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WO2020020183A1 - Novel bamboo salt and preparation method therefor - Google Patents

Novel bamboo salt and preparation method therefor Download PDF

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Publication number
WO2020020183A1
WO2020020183A1 PCT/CN2019/097358 CN2019097358W WO2020020183A1 WO 2020020183 A1 WO2020020183 A1 WO 2020020183A1 CN 2019097358 W CN2019097358 W CN 2019097358W WO 2020020183 A1 WO2020020183 A1 WO 2020020183A1
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WO
WIPO (PCT)
Prior art keywords
salt
plant
bamboo
extract
functional
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Application number
PCT/CN2019/097358
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French (fr)
Chinese (zh)
Inventor
张英
陈亮
周沫希
严雅丽
黄骆镰
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杭州尤美特科技有限公司
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Priority to KR1020217005372A priority Critical patent/KR20210061998A/en
Publication of WO2020020183A1 publication Critical patent/WO2020020183A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/40—Table salts; Dietetic salt substitutes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/10—General methods of cooking foods, e.g. by roasting or frying
    • A23L5/15—General methods of cooking foods, e.g. by roasting or frying using wave energy, irradiation, electrical means or magnetic fields, e.g. oven cooking or roasting using radiant dry heat
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/14—Alkali metal chlorides; Alkaline earth metal chlorides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/15—Inorganic Compounds
    • A23V2250/156—Mineral combination
    • A23V2250/1638—Undefined mineral extract

Definitions

  • the invention relates to the technical field of food and health food, in particular to a functional plant salt and a preparation method thereof, and more particularly to a bamboo salt and a preparation method thereof.
  • Plant salt refers to a kind of balanced salt extracted from plant organisms, using pure physical processes and processes, conforming to low sodium salts and a variety of mineral trace element groups, and various element proportions meeting the requirements of the human body. , A new type of edible salt with a certain promoting effect on function regulation and metabolism and health.
  • Plants are the main source; 2 Natural low sodium salt and natural balanced salt; 3 Rich in various organic nutrients and biologically active ingredients; 4 No chemical synthetic substances are added during processing and production; 5 A variety of diseases and related complications caused by the lack or imbalance of mineral element groups have certain adjuvant treatment and regulating effects. Such as sodium-sensitive hypertension, type 2 diabetes, hyperlipidemia, cardiovascular disease, tumors and immune dysfunction and other diseases and complications have a clear effect.
  • Plant-Salt Plant-Salt
  • the technical problem to be solved by the present invention is to provide a functional plant salt and a method for preparing the functional plant salt.
  • the functional plant salt is bamboo salt, which refers to a product prepared by using bamboo extract as a plant-based raw material.
  • a functional plant salt is provided.
  • the functional plant salt is prepared by mixing one or more plant extract powders with fine salt in an amount of 1% to 40%. Uniform, roasted once or in multiples at a high temperature of 800-1400 ° C to obtain a functional plant salt.
  • the plant extract is a bamboo extract.
  • the plant extract is selected from the group consisting of bamboo leaf extract, bamboo rutberry extract, bamboo pole extract, bamboo shoot extract, or a combination thereof.
  • the plant extract is selected from the group consisting of bamboo leaf flavones, bamboo leaf polyphenols, or a combination thereof.
  • the fine salt is selected from the group consisting of sea salt, lake salt, well salt, and mineral salt.
  • the fine salt is crude sea salt.
  • the functional plant salt is bamboo salt.
  • the plant extract further comprises a plum extract and / or a pine extract.
  • the fine salt is common salt.
  • the ⁇ L value of the functional plant salt is 28-50, preferably 35-50.
  • the ⁇ L value of the functional plant salt is 30-40, preferably 32-39.
  • a method for preparing a functional plant salt according to the first aspect of the present invention comprising the steps of: adding one or more plant extract powders in an amount of 1% to 40% and Fine salt is mixed uniformly, and it is roasted once or in multiples at a high temperature of 800-1400 ° C to obtain a functional plant salt.
  • the roasting is performed in a mechanized oven.
  • the roasting is performed in an intermediate frequency melting furnace.
  • a food or pharmaceutical composition comprising one or more functional plant salts according to the first aspect of the present invention.
  • a fourth aspect of the present invention there is provided the use of the functional plant salt according to the first aspect of the present invention, for preparing a substance selected from the group consisting of a pharmaceutical auxiliary, a health food / drink, and a functional food / drink , Table seasoning salt, cooking salt, special purpose cosmetics or personal care products.
  • a method for inhibiting transaminase activity comprising the step of applying one or more inhibitory effective amounts of the functional plant salt according to the first aspect of the present invention to a desired patient.
  • aminotransferase is selected from the group consisting of aspartate aminotransferase (ALT) and glutamic acid aminotransferase (AST).
  • FIG. 1 is a photograph of the pine salt obtained in Example 4.
  • FIG. 1 is a photograph of the pine salt obtained in Example 4.
  • FIG. 2 is a photograph of bamboo salt obtained in Example 5.
  • FIG. 2 is a photograph of bamboo salt obtained in Example 5.
  • FIG. 3 is a photograph of plum salt obtained in Example 6.
  • FIG. 4 is a photograph of the pine salt obtained in Example 7.
  • FIG. 4 is a photograph of the pine salt obtained in Example 7.
  • FIG. 5 is a photograph of bamboo salt obtained in Example 8.
  • FIG. 6 is a photograph of plum salt obtained in Example 9.
  • FIG. 7 is a photograph of plant salts obtained under different formulations and different process parameters.
  • Figure 8 is a micrograph of different salts.
  • Figure 9 is the SEM results of different salts.
  • Figure 10 is the TEM results of different salts.
  • Figure 11 is the XRD results of different salts.
  • Figure 12 shows the results of the electronic tongue test for different salts.
  • Figure 13 shows the results of electronic nose tests for different salts.
  • FIG. 14 is a morphological observation result of pathological sections of liver tissue of mice in each group.
  • Figure 17 is the effect of 0.5% mass fraction of each sample on the migration of B16 cells, a-normal; b-original salt; c-arbutin; d-pine salt; e-bamboo salt; f-plum salt.
  • Figure 18 shows the effect of 0.5% mass fraction of each sample on the B16 cell cycle, a-normal; b-raw salt; c-arbutin; d-pine salt; e-bamboo salt; f-plum salt.
  • FIG. 19 is a graph showing changes in body weight of experimental animals in 6.3.1.
  • Figure 20 shows the changes in blood pressure in experimental rats in 6.3.2.
  • the inventors After a long and intensive study, the inventors have unexpectedly prepared a kind of high-quality treatment by using a mixture of unpre-roasted bamboo extract and raw salt at a temperature higher than the melting point of the raw salt (such as 800-1400 ° C). Biological effects of bamboo salt.
  • the preparation method of the bamboo salt is simple and easy to popularize on a large scale. On this basis, the inventors have completed the present invention.
  • the term "functional plant salt” is bamboo salt, which refers to a product prepared by using bamboo extract as a plant-based raw material.
  • the term "without pre-baking" means that the raw material for preparing the bamboo salt is a direct mixture of bamboo extract and raw salt, and the mixture has not been subjected to one or more low temperatures (such as below 800 ° C) of traditional preparation of bamboo salt. ) Pre-baking treatment.
  • plant extract is a bamboo extract.
  • the bamboo extract is selected from the group consisting of bamboo leaf extract, bamboo rut extract, bamboo pole extract, bamboo shoot extract, or a combination thereof.
  • the bamboo extract is selected from the group consisting of bamboo leaf flavones, bamboo leaf polyphenols, bamboo triterpenes, bamboo polysaccharides (such as bamboo polysaccharides), bamboo shoot sterols, bamboo shoot amino acid peptides, or combinations thereof .
  • the well-known Korean bamboo salt actually originated in China. According to legend, more than 1300 years ago, a caravan stayed at a temple on the Ancient Tea Horse Road. At night, the horse stables caught fire, and the white table salt and the container (bamboo basket) containing salt were burned into gray salt blocks (that is, the prototype of bamboo salt). The monks used it to treat people's indigestion, trauma, hemostasis, and disinfection. The effects were significant and gradually spread. So far, in some temples in Yunnan and some mountainous areas in Zhejiang province, some people still put salt in bamboo tubes and calcined them in the stove to eat.
  • bamboo salt has certain anti-inflammatory, anti-allergic, anti-cancer, antibacterial, prevent obesity, protect teeth, anti-gastric ulcer, liver protection, anti-aging, prevent hypertension, prevent blood acidification, Improve diabetes, treat arthritis, and enhance immunity.
  • Traditional bamboo salt production uses sun-sea salt as raw material, which is filled into fresh bamboo bamboo (that is, moso bamboo), sealed with loess, pine wood as fuel, and repeatedly baked in an earth kiln. According to the filling, The number of roasts distinguishes between quality and grade.
  • roasted bamboo salt needs to repeat 9 cycles of "filling-roasting-cooling-pulverizing-refilling", the purpose is to let the salt absorb more beneficial ingredients from the fresh bamboo tube; and during the last roasting, Pine resin is added to pine wood to make the baking temperature reach 1300 ° C. After cooling, blue-violet salt is obtained, which is called purple bamboo salt.
  • a production cycle takes about one month.
  • a well-known domestic bamboo salt manufacturer (Sanhe Garden, Lin'an, Zhejiang) described the traditional roasting process as follows: remove moisture at 100-300 ° C, and burn the raw salt and organic matter in the bamboo tube at 300-800 ° C; 800-1200 ° C Some of the heavy metal elements in the time salt are distilled away in the form of chloride, and the trace elements in bamboo tube and yellow mud are combined with sodium chloride; the molten table salt is taken out at a high temperature above 1300 ° C, and rapid cooling causes 1 crystal. Distortion of the grid can get bamboo salt.
  • the melting point of NaCl is 801 ° C.
  • the temperature for melting the salt should reach at least about 900 ° C.
  • the effective ingredients of bamboo that can be absorbed in this way are very limited, so it needs to be recycled repeatedly. For example, nine roasted bamboo salt (purple bamboo salt) needs to be recycled nine times.
  • the traditional pine wood-fired kiln can generally reach a temperature of about 800 ° C, in order to finally melt the salt, the method of adding turpentine to pine branches during the ninth roasting to increase the furnace temperature, Claimed to be able to reach more than 1300 ° C (the lack of support of measured data, it is estimated that the actual temperature can reach about 900 ° C).
  • bamboo salt has been a niche product and cannot reach more consumers.
  • the invention discloses a class of functional plant salts, a preparation method and uses thereof, and relates to the fields of food, condiments, daily chemicals, and medicines, and the purpose is to provide a natural, safe, nutritious, and multiple biological health care effect.
  • the core is to use intermediate frequency melting technology or resistance heating technology. Mix different types of plant extracts from different sources with edible salt in a certain mass ratio (0-40%) and put them into an intermediate frequency melting furnace or resistance furnace. The temperature is reduced to 800 to 1400 ° C. and melted, and then crystallized after cooling, so as to obtain plant salt series with different colors, different physical and chemical properties, different lattice structures, different flavors, and different uses.
  • the salt solution changes from weak acidity to strong alkalinity, and the color changes from white to gray, light gray, pale pink, fuchsia, and bright blue and other colors, showing a clean and clear appearance with a touch of light
  • the sulphur taste of the salt is smaller than that of the original salt, and the elemental composition is more abundant.
  • the potassium content is increased by about 100 times, the calcium is about 7 times, and other beneficial elements are also significantly increased.
  • the health benefits of functional plant salts include, but are not limited to: whitening, liver protection, stomach strengthening, detoxification, anti-inflammatory, anti-allergy, oral cleansing, prevention of hypertension, treatment of migraine, inhibition of intestinal canceration, and can be widely used in food, Beverages, condiments, health foods, medicines, daily chemicals, personal care products, etc. have a bright future. Since the melting point of the sodium chloride crystal lattice is 801 ° C, the temperature for completely melting the table salt is at least about 900 ° C, and the traditional furnace cannot generally meet this temperature requirement.
  • Intermediate frequency melting and resistance heating are two technologies that can efficiently convert electrical energy into thermal energy, which can quickly reach melting temperatures between 900 and 1700 ° C, have high production efficiency, and can achieve mechanization, automation, scale, standardization and clean production .
  • functional plant salt series with different colors, stable performance, excellent quality, and standardizable quality can be obtained by controlling the type of plant extracts, the addition ratio, the smelting temperature, the holding time, and the heating and cooling rates.
  • the invention creatively applies intermediate frequency melting technology or resistance heating technology, and after mixing table salt with different plant extracts, it is smelted at a high temperature of 800 to 1400 ° C to form different colors, different lattice structures, different physical and chemical indexes, Functional plant salt series with different flavors and uses.
  • Intermediate frequency smelting is a commonly used metal smelting method, that is, to convert AC power at 50Hz to intermediate frequency (300 ⁇ 1000Hz), rectify the three-phase power frequency AC to DC, and then convert DC to adjustable intermediate frequency current.
  • Intermediate frequency melting technology can reach the required melting temperature in a short time, greatly improving production efficiency.
  • the resistance heating technology converts electrical energy into heat energy through a resistor body, thereby rapidly heating the material.
  • the resistance furnaces currently in commercial use in China mainly use molybdenum rods, silicon rods or carbon rods as heating elements, and the maximum furnace temperature can reach 1700 ° C.
  • Resistance heating technology can control heating time, temperature, heating rate, etc., so as to achieve precise control of process parameters and standardization of products.
  • Plant extracts also known as phytochemicals (that is, plant secondary metabolites), refer to the use of different technical means to extract, isolate, and purify from various fruits, vegetables, flowers, Chinese herbal medicine, medicinal and edible plants, and new resource foods.
  • Different precision formulations and even characteristic monomer compounds such as extracts from plum, orchid, bamboo, chrysanthemum, pine, lotus, tea, pomegranate, begonia, edible fungi, algae and the like.
  • the raw material of the algae extract can be seaweed (such as kelp, wakame, sargassum, etc.), or algae cultivated in freshwater or seawater, such as cyanobacteria, green algae, chlorella, etc .
  • edible mushroom extracts include shiitake mushrooms , Mushrooms, enoki mushrooms, bamboo mushrooms, mushrooms, fungus, white fungus, monkey head, ganoderma, truffle, boletus, etc. and their processing residues (such as mushroom handles, etc.);
  • microbial-derived extracts include various types of yeast Extracts (also known as yeast extract), such as beer yeast, baker's yeast, and the like.
  • the main technical features of the present invention are: after using intermediate frequency melting technology or resistance heating technology, plant extracts from different sources are mixed with table salt at a ratio of 0 to 40% (the extract powder will be evenly adsorbed on the surface of the table salt particles) , Under normal pressure, the temperature was raised to 800 ⁇ 1400 °C, and then smelted. After cooling, it was recrystallized to obtain plant functional salt series with different colors, different lattice structures, different physical and chemical indexes, different flavors and different uses.
  • the new plant salt series has whitening, liver protection, stomach strengthening, detoxification, anti-inflammatory, anti-allergy, oral cleansing, and prevention of hypertension .
  • Multiple functions such as treating migraine and inhibiting intestinal cancer, can be widely used in food, beverages, condiments, health foods, pharmaceuticals, daily chemicals, personal care products and other fields.
  • the heating rate is 1 ⁇ 200 °C / min; 1 ⁇ 20 °C / min;
  • the heating temperature can be set in one or more intervals between 800 and 1400 ° C as required;
  • the above mixing-melting-recrystallization-cooling-pulverizing process can be performed one or more times.
  • the raw common salt (also called raw salt) according to the present invention may be sea salt, lake salt, well salt, mineral salt, and preferably crude sea salt (that is, sun salt).
  • the functional plant salt provided by the present invention according to different sources of plant extract, addition ratio, melting temperature and other parameters, its health effect can be specifically expressed as non-bamboo functional plant salt (such as pine salt, plum, etc.)
  • the health benefits of salt include, but are not limited to, whitening, liver protection, and prevention of hypertension. Its applications include food, beverages, condiments, health foods, pharmaceuticals, household chemicals and personal care products.
  • the intermediate frequency smelting technology and the resistance heating technology of the present invention can also be directly used for processing table salt, that is, without adding any plant extract or other foreign substances in the manufacturing process, the raw salt is converted into a food with higher safety and appearance More crystal clear grilled salt.
  • the outstanding advantage of the present invention is that by using intermediate frequency melting technology or resistance heating technology, raw salts can be mixed with plant extracts (powder) of any source and ratio, and recrystallized after co-melting in a precisely set temperature interval, and once through Or it can be roasted several times to obtain salt with special biological effects, and it can realize large-scale industrial production of mechanization, automation, standardization and cleanliness.
  • the invention creates non-bamboo series plant functional salt represented by pine salt and plum salt, which is a revolution to the table salt industry (especially the high-end salt market) and is of great significance to the health cause of all human beings .
  • the present invention proposes a method for preparing a functional plant salt, which includes the following steps:
  • One or more plant extract powders are mixed with fine salt uniformly at an added amount of 1% to 40%, and roasted at a high temperature of 800-1400 ° C once or in stages to obtain a functional plant salt.
  • a plant extract powder is uniformly mixed with fine salt at an addition amount of 1% to 40%, and is roasted at a high temperature of 800-1400 ° C at one time to obtain a functional plant salt.
  • the obtained mixed powder is uniformly mixed with fine salt at an addition amount of 1% to 40%, and is roasted at a high temperature of 800-1400 ° C at one time to obtain a functional plant salt.
  • step S2 Take out the product that was kept at 800-1400 ° C for a certain time in step S1, add another plant extract powder to it after crushing, and repeat steps S1 and S2 (Note: at this time, the original temperature or higher Continue to roast) until all the plant extract powder is roasted to obtain a functional plant salt.
  • the present invention also provides a functional plant salt prepared by using the above preparation method.
  • the present invention also proposes the use of a functional plant salt:
  • the functional plant salt is used in pharmaceutical auxiliaries, health foods / drinks, functional foods / beverages, table seasoning salts, cooking salts, special-purpose cosmetics or personal care products.
  • the functional plant salt according to the present invention is characterized in that, by using intermediate frequency melting technology or resistance heating (also known as ohmic heating) technology, bamboo extracts from different sources are mixed with table salt in a certain ratio,
  • the new bamboo salt obtained by smelting at 800 ⁇ 1400 °C and recrystallizing after cooling can be widely used in food, beverages, condiments, health foods, pharmaceuticals, daily chemicals, personal care products, etc.
  • the non-bamboo plant functional salt represented by pine salt and plum salt which is pioneered by the present invention, has the same biological effects as bamboo salt, and also has various functions such as preventing hypertension, whitening, and protecting the liver.
  • the salt solution changes from weakly acidic to strongly alkaline, and the color changes from white to gray, light gray, light pink, fuchsia, and bright blue, showing a clean and clear appearance, accompanied by There is a slight sulphur smell; the unit cell size of the salt is smaller than that of the original salt, and the elemental composition of the salt is more abundant, in which the potassium content is increased by about 100 times, calcium is about 7 times, and other beneficial elements are also significantly increased.
  • the intermediate frequency melting technology is to convert AC power of 50Hz to intermediate frequency (300 ⁇ 1000Hz), rectify the three-phase power frequency AC to DC, and then convert DC to adjustable intermediate frequency current.
  • the intermediate frequency alternating current flowing in the induction coil generates high-density magnetic lines of force in the induction coil, and rapidly produces high temperature through a magnetically permeable container (such as a graphite silicon carbide crucible) to heat the material (that is, a mixture of salt and extract) Let it melt.
  • the melting temperature can be reached quickly in a short time, which greatly improves the production efficiency.
  • the melting temperature can be set in one or more intervals between 800 and 1400 ° C;
  • the above mixing-melting-cooling-recrystallization-pulverizing process can be performed one or more times.
  • Resistance heating also known as ohmic heating
  • ohmic heating converts electrical energy into thermal energy to rapidly heat materials.
  • the ratio of resistance furnace to flame furnace has high thermal efficiency and easy temperature control. It is divided into direct heating and indirect heating.
  • most resistance furnaces use indirect heating, and the equipment is equipped with a resistance element that realizes electric-thermal conversion, called an electric heating body, which transfers heat energy to the material to be processed.
  • Commonly used electric heating bodies are silicon carbide rods and molybdenum disilicide rods.
  • the temperature of silicon carbon rods can reach 1400 ° C, and silicon molybdenum rods can reach 1700 ° C.
  • the heating rate is 1 ⁇ 20 °C / min;
  • the melting temperature can be set in one or more intervals between 800 and 1400 ° C;
  • the above mixing-melting-recrystallization-cooling-pulverizing process can be performed one or more times.
  • the plant extract also known as a phytochemical, is a secondary metabolite of a plant. Contains a variety of fruits, vegetables, flowers, Chinese herbs, medicinal and edible plants, and plant-derived new resource foods, crude plant extracts, high-precision preparations and even monomer compounds such as bamboo, plum, pine, tea, lotus, chrysanthemum , Pomegranate, begonia, edible fungus, algae extract, etc.
  • the same plant source can also be extracts from different parts of it, such as bamboo extracts including bamboo leaf extracts (such as bamboo leaf flavones, bamboo leaf polyphenols), bamboo rut extracts (such as ruta triterpenes, ruta polysaccharides), bamboo pole extract, bamboo shoot extract (such as bamboo shoot sterol, bamboo shoot amino acid peptide), plum extracts include plum fruit extract, plum extract, plum leaf extract, plum branch extract, plum root extract, plum kernel extract, pine The extracts include pine needle extract, pine bark extract, and pine pollen extract.
  • bamboo extracts including bamboo leaf extracts (such as bamboo leaf flavones, bamboo leaf polyphenols), bamboo rut extracts (such as ruta triterpenes, ruta polysaccharides), bamboo pole extract, bamboo shoot extract (such as bamboo shoot sterol, bamboo shoot amino acid peptide), plum extracts include plum fruit extract, plum extract, plum leaf extract, plum branch extract, plum root extract, plum kernel extract,
  • plant functional salts are divided into two series: bamboo and non-bamboo. It is classified as bamboo functional salt by using bamboo extract series.
  • Non-bamboo functional salts are collectively referred to as non-bamboo functional salts. They include algae, mushrooms (edible fungi) and microbial-derived extracts (or extracts). Functional salt.
  • the common salt raw material (that is, raw salt) may be sea salt, lake salt, well salt, mineral salt, preferably crude sea salt (also known as sun-sea salt).
  • the health effects of the non-bamboo functional salts include, but are not limited to, preventing hypertension, whitening, and protecting the liver.
  • the application fields of the functional plant salt include: food, beverages, condiments, health foods, medicines, daily chemicals and personal care products.
  • the intermediate frequency melting technology or the resistance heating technology of the present invention can also directly process table salt (that is, without adding plant extracts or any other foreign substances), and convert it into roasted salt with higher food safety.
  • the present invention has the following technical advantages:
  • the specific biological functions of the functional plant salt prepared by the present invention include: bacteriostatic, anti-inflammatory, anti-viral; protecting the liver and kidney, soothing the stomach, detoxifying and detoxifying, stopping bleeding and hemorrhoids; treating migraine, allergic rhinitis, Anti-inflammatory effect on human gingival fibroblasts, weight loss function, protecting oral health, inhibiting colon cancer, increasing anti-mutation and anti-cancer effects in vitro, anti-mutation activity of HepG2 human liver cancer cells and anti-cancer effects in vitro, preventing neuronal apoptosis Wait. In addition, it helps to improve the permeability of human epidermal cells and helps skin care factors in daily chemicals work better.
  • the functional plant salt prepared by the present invention include: pharmaceutical auxiliaries; health food / drinks; functional foods / drinks; table seasoning salt, cooking salt; special-purpose cosmetics, personal care products (skin cream, liquid foundation, soap , Facial mask, toothpaste, mouthwash, shampoo, bath salt, etc.); the form of the end product can be powder, capsule, tablet, granule, effervescent tablet, water, emulsion, spray, etc.
  • Zhuru triterpenes yellow-green powder, provided by the "Natural Products and Human Health” research center of the College of Biological System Engineering and Food Science, Zhejiang University, with a total triterpenoid saponin content of 45%;
  • Pine needle extract brown powder, purchased from Shaanxi Qingya Biotechnology Co., Ltd .;
  • Plum branch extract yellow powder, provided by Hangzhou Youmeite Biotechnology Co., Ltd .;
  • Raw salt (sun-sea salt): from Weifang, Shandong, the measured NaCl content is 93.36%;
  • the heating element is a copper induction coil
  • the crucible is a magnetically conductive graphite silicon carbide crucible
  • Tube resistance furnace The heating element is a silicon carbon rod.
  • Example 1 A method for preparing a functional plant salt, including the following steps:
  • plant extracts include, but are not limited to, plant extracts of various fruits, vegetables, edible flowers, Chinese herbal medicine and medicinal food plants, as well as new sources of food derived from plants, such as bamboo, plum, pine, tea, lotus, Daisy, pomegranate, begonia, edible fungus, algae extract, etc.
  • the same plant source can also be extracts from different parts of it, such as bamboo extracts including bamboo leaf extracts (bamboo leaf flavones, bamboo leaf polyphenols), bamboo rut extracts, bamboo pole extracts, bamboo shoot extracts, and plum extracts including Plum blossom extract, plum fruit extract, plum leaf extract, plum branch extract, plum root extract, pine extracts include pine needle extract, pine bark extract, pine pollen extract, and the like.
  • bamboo extracts including bamboo leaf extracts (bamboo leaf flavones, bamboo leaf polyphenols), bamboo rut extracts, bamboo pole extracts, bamboo shoot extracts, and plum extracts including Plum blossom extract, plum fruit extract, plum leaf extract, plum branch extract, plum root extract, pine extracts include pine needle extract, pine bark extract, pine pollen extract, and the like.
  • step 2 Mix the plant extract powder in step 1 with the fine salt in an amount of 1% to 40% to obtain a mixture;
  • the obtained mixture is put into a special container, and is heated to 800-1400 ° C. in a medium-frequency melting furnace (or other mechanized oven) at a certain heating rate to obtain a functional plant salt.
  • a medium-frequency melting furnace or other mechanized oven
  • the source of the fine salt may be sea salt, lake salt, well salt, mineral salt, and preferably crude sea salt.
  • the above% is a mass ratio.
  • Example 2 The plant extract powder in Example 1 was changed to at least two plant extract powders, that is, the at least two plant extract powders were mixed uniformly at any ratio to obtain a mixed powder; the mixed powder was 1% The added amount of -40% is mixed with fine salt (sieved) uniformly to obtain a mixture; the rest is equivalent to Example 1.
  • Embodiment 3 The plant extract powder in Example 1 is changed to at least two plant extract powders (one of which is bamboo extract), and the specific steps are as follows:
  • step 3 Take out the product that was kept at 800-1400 °C for a certain period of time in step 2 and add another plant extract powder to it after crushing, and repeat steps 1-3 (Note: at this time at the original temperature or higher Continue to roast) until all plant extract powder is roasted to obtain functional plant salt
  • the total added amount of the plant extract powder used above is 1% to 40% (mass ratio).
  • the functional plant salt of the present invention can be roasted at one time after adding plant extracts, or it can be roasted in stages and added in stages.
  • the elemental composition, NaCl crystal lattice, and crystal configuration have undergone significant changes.
  • different colors are displayed, such as gray, purple, pink, white, blue, turquoise, etc .
  • the pH value of the 5% saline solution is increased from 6-7 of the raw salt to 8-12
  • the total amount of NaCl decreases, and the content of other elements (especially a variety of trace elements) increases; the taste changes from a simple salty taste to a rich and rounded savory sweetness, and it also has a significant rotten egg smell.
  • the "pine salt”, “bamboo salt”, and “plum salt” obtained after adding pine, bamboo, and plum extracts are different due to the origin of the extract, the chemical composition and the amount of addition, and the number of roasting times and process parameters
  • the physical and chemical properties, appearance, color, and flavor of the obtained plant salt are significantly different, and the biological effects caused by them are also different.
  • the specific biological functions of functional plant salts include: bacteriostatic, anti-inflammatory, anti-viral; liver-protecting kidney, soothe the stomach, detoxify and detoxify alcohol, stop bleeding and relieve hemorrhoids; treat migraine, allergic rhinitis, and affect human gums
  • Fibrocytes have anti-inflammatory effects, weight loss functions, protect oral health, inhibit colon canceration, increase anti-mutation and anti-cancer effects in vitro, anti-mutation activity of HepG2 human liver cancer cells and anti-cancer effects in vitro, and prevent neuronal apoptosis. In addition, it helps to improve the permeability of human epidermal cells and helps skin care factors in daily chemicals work better.
  • functional plant salts include: as pharmaceutical auxiliaries; health foods / drinks; functional foods / drinks; table seasoning salts, cooking salts; special-purpose cosmetics, and human care products (skin creams, liquid foundations, soaps, masks , Toothpaste, mouthwash, shampoo, bath salt, etc.); the form of the end product can be powder, capsule, tablet, granule, effervescent tablet, water, emulsion, spray and so on.
  • the present invention provides a method for preparing a functional plant salt.
  • Different plant extracts and table salt are mixed uniformly at a certain ratio, and they are formed by roasting one or several times at a high temperature of 800-1400 ° C.
  • Functional plant salts of different colors, different spatial structures, different physical and chemical properties, different flavors and different uses.
  • the functional plant salt prepared by the invention not only has a health-care effect, but also helps to improve the permeability of human epidermal cells, and helps the skin care factors in daily chemicals to function better. It can be used in pharmaceutical auxiliaries, health foods / Beverages, functional foods / drinks, table salt, cooking salt, special purpose cosmetics or care products.
  • Figure 7 shows raw salt added with 5 to 35% of different plant extracts, subjected to high temperature smelting at 800 to 1400 ° C for 1 to 2 hours under normal pressure, the pine salt obtained in Example 4, the bamboo salt obtained in Example 5, and Example 6
  • the appearance of the obtained plum salt can show different colors such as gray, light gray, pale pink, blue-violet and bright blue.
  • the roasted salt obtained by smelting the raw salt without adding any foreign substances at a high temperature of 800 to 900 ° C. for 1 to 2 hours has higher transparency and brighter color.
  • the color difference meter is a common photoelectric integrating color measuring instrument. It imitates the principle of color perception of the human eye. It uses a light receiver that can sense three colors of red, green, and blue. The standard light source inside the instrument is used to illuminate the measured object. The photocurrents experienced by each are amplified, and an integral measurement is performed over the entire visible light wavelength range to obtain the tristimulus value and chromaticity coordinates of the color of the transmitted or reflected object, thereby obtaining the signal of this color, and giving the signal by the system. Measure the color difference between samples.
  • the main indicators measured by the color difference meter include L *, a *, b *. among them,
  • L * represents the brightness of the sample, the larger the value, the brighter it is
  • a * stands for red and green, "+” means red, and "-” means green.
  • b * represents a blue-yellow phase
  • "+” indicates a bias toward yellow
  • "-” indicates a bias toward blue
  • the ⁇ L value represents the difference between the sample value and the reference point;
  • the ⁇ a value represents the difference between the a * value of the sample and the reference point, which can better represent the red-green value of the sample.
  • the reference point refers to the value of the standard sample
  • the present invention uses a standard white board as the standard sample.
  • Table 1 is a summary of the color difference analysis data of different plant functional salts. It can be seen that the ⁇ L values of raw salt and roasted salt are 84.18 ⁇ 0.25 and 82.83 ⁇ 3.47, showing that they are close to white; while the ⁇ L values of other plant functional salts are between 25-50, the darkest is the mangosteen salt, the brightest It's plum salt. The a * value and b * value of different samples are significantly different (p ⁇ 0.05), which indicates that the color direction of them differs greatly, showing a completely different hue.
  • the pH value of food has an important effect on maintaining the acid-base balance of the human body environment, so there is a distinction between acidic foods and alkaline foods.
  • the dietary structure of the general public is generally acidic. Over time, it will form an acidic constitution, which will lead to a decline in immunity and a sub-healthy state.
  • the pH values of different salt samples were measured at a concentration of 0.2 to 5% (w / v), as shown in Table 2.
  • the original salt solution with a concentration of 1% or less is weakly acidic (pH ⁇ 7), and it rises significantly after baking; all the plant functional salt solutions show strong alkalinity (pH> 9) even at a concentration of 0.2%, which is in phase with the original salt. The difference is very significant (p ⁇ 0.001). It is shown that after the original salt is fused with the plant extract, a large number of alkali metal elements have entered the lattice structure of NaCl or adhered to the surface.
  • Atomic energy spectroscopy can not only scan and photograph the surface morphology of materials, but also detect various elements on the surface of materials and their relative contents.
  • the plant functional salt prepared by the present invention and its reference sample (particles) are placed under an atomic energy spectrometer (Hitachi, Japan, SU-8010) to perform element qualitative, semi-quantitative, and quantitative analysis on the micro-area points, lines, and surfaces of the sample Analysis (detection limit: 0.5%), the results are shown in Table 4.
  • the main ingredient of salt is sodium chloride, and it also contains small amounts of other mineral elements.
  • the more refined the salt the higher the sodium chloride content, the less other nutrients, which is not good for human health.
  • Potassium can regulate intracellular osmotic pressure and body fluid acid-base balance, participate in the metabolism of sugar and protein in the cell, help maintain nerve health, normal heartbeat, prevent stroke, and assist normal muscle contraction. In the case of high blood pressure caused by ingestion of high sodium, potassium or potassium-rich foods have a hypotensive effect.
  • Magnesium is the main cation in human cells. It can affect the transport of potassium and calcium ions, regulate signal transmission, participate in energy metabolism, protein and nucleic acid synthesis, and activate and inhibit catalytic enzymes.
  • Phosphorus is one of the basic components of nucleic acids that make up genetic material. At the same time, phosphorus and calcium are important building materials for bones and teeth. Phosphorus can maintain the balance of ATP metabolism in the body, regulate the acid-base balance in the body, and participate in the metabolism of energy in the body.
  • Molybdenum is an essential trace element for the human body. It is one of the basic components of xanthine oxidase and aldehyde oxidase in the liver and intestines of animals. It is also a component of heparin sulfase oxidase. Studies have shown that molybdenum also has a significant anti-caries effect, and has a strong inhibitory effect on the formation of urinary stones. Humans lacking molybdenum are prone to kidney stones.
  • Vanadium is an essential trace element in the human body. It is generally believed that it may help prevent cholesterol accumulation, lower blood sugar, lower blood pressure, prevent dental caries, and help make red blood cells.
  • Selenium is an essential trace element necessary for the human body.
  • the Chinese Nutrition Association lists selenium as one of the 15 essential nutrients for the human body. It is generally believed that selenium is closely related to immunity, aging, and reproductive functions. Selenium deficiency is an important cause of Keshan disease.
  • Chromium is also an essential micronutrient, which plays an important role in all insulin regulating activities and is an important blood glucose regulator. At the same time, the lack of chromium is also related to the formation of myopia.
  • Silicon is mainly concentrated in bones, lungs, lymph nodes, pancreas, adrenal glands, nails, and hair. It is the highest in connective tissues such as aorta, trachea, tendons, bones, and skin, and has special significance for human bone health. However, due to industrial development and changes in lifestyle, the silicon element in modern human bodies is often extremely insufficient.
  • plant functional salt and original salt have very significant differences.
  • the potassium content of pine, bamboo, and plum salts has increased significantly, especially the potassium content of the new bamboo salt obtained in Examples 2 and 3 is more than 100 times that of the original salt, and it is 14.9% higher than that of Renshan bamboo salt produced by traditional processes;
  • the salt (including the old and new samples) contained magnesium, iron, nickel, zinc, molybdenum, and vanadium significantly higher than the original salt and other plant salts.
  • the selenium was also detected in the pine and plum salts obtained by the new technology. .
  • plum salt had much higher iodine (28mg / kg), boron (26.6mg / kg), phosphorus (424mg / kg), chromium (4mg / kg) and second only to bamboo salt than other samples.
  • Manganese content (18mg / kg).
  • the silicon content of plant salt is significantly higher than that of raw salt and roasted salt, and the bamboo salt of the new process is higher than the traditionally produced Renshan bamboo salt.
  • selenium was detected in pine and plum salts.
  • Hydrogen sulfide has a variety of neuroprotective effects, which have been confirmed by a large number of experiments at home and abroad, mainly including anti-neuroinflammatory response, anti-oxidative stress injury and anti-hypoxic-ischemic nerve injury, which can increase the plasticity of nerve cells and reduce hippocampus Neuronal apoptosis.
  • the detection limit of phosphorus is 20mg / kg; the detection limit of molybdenum, antimony, selenium, tin, and total arsenic is 0.01mg / kg; the detection limit of cadmium and * germanium is 0.001mg / kg; the detection of radon The limit is 0.0001 mg / kg.
  • a stereo microscope (Jiangnan Brand SE220) was used to capture the appearance of the sample of the plant functional salt particles obtained during the implementation under the conditions of 10 times the eyepiece and 5 times the objective lens, as shown in FIG. 8.
  • the raw salt and roasted salt are basically white, the particle surface is relatively smooth, and the shape is regular.
  • pine salt, bamboo salt, and plum salt show different colors, and the crystal arrangement differs greatly, giving them a more lustrous feel.
  • the color of the new bamboo salt prepared by the present invention is similar to that of Renshan bamboo salt, but the former is more pure and looks crystal clear, while the latter has more impurities.
  • the GeminiSEM 300 scanning electron microscope produced by Carl Zeiss Microscopy GmbH of Germany was used to perform morphological characterization of the plant functional salts of the examples.
  • the different sample particles were respectively observed under an electron microscope, and FIG. 9 was obtained.
  • the surface of raw salt and roasted salt is smooth without much attachment.
  • the surface of the plant functional salt prepared by the present invention is in the shape of a sweet potato, and many small particulate materials are attached to the particle surface.
  • the pine salt obtained in Example 4, bamboo salt obtained in Example 5, plum salt obtained in Example 6 and its control sample were ground into powder (passed through a 100-mesh sieve), and an X-ray diffractometer (Bruker D8 Advance) produced by Bruker, Germany was used. Perform analysis. Since the crystal is composed of unit cells arranged regularly, these X-rays scattered by different atoms interfere with each other, resulting in strong X-ray diffraction in some special directions. The orientation and intensity of the diffraction line in spatial distribution are closely related to the crystal structure. The diffraction pattern produced by each crystal reflects the atomic distribution of the crystal. The sample can be analyzed for phase analysis, unit cell parameters, and diffraction line intensity analysis.
  • Phase analysis is the most commonly used aspect of X-ray diffraction in metals and is divided into qualitative and quantitative analysis.
  • the former compares the lattice plane spacing and diffraction intensity measured on the material with the diffraction data of the standard phase to determine the phase present in the material.
  • Accurate unit cell parameter data can reflect the slight differences in structure between different samples of a substance, or the small changes in the structure of a crystal caused by external physical and chemical factors.
  • the unit cell parameters of several phases of the plant functional salt obtained in the examples are as follows:
  • the unit cell parameters of different salt samples were simulated by unit cell parameters. From Figure 11, it can be seen that the raw salt is more regular and cubic, while the ends of the pine salt are sharper, the bamboo salt is more round, and the plum salt has an octahedral cone shape. The crystal structure has changed a lot.
  • the electronic tongue is to simulate the human tongue to analyze, identify and judge the sample to be tested, and use multivariate statistical methods to process the obtained data, quickly reflect the overall quality information of the sample, and realize the identification and classification of the sample.
  • It is a detection technology that uses the multi-sensor array as the basis to sense the overall characteristic response signal of the sample, and performs analog identification and quantitative qualitative analysis on the sample. It is mainly composed of taste sensor array, signal acquisition system and pattern recognition system.
  • Electronic nose also known as odor scanner
  • odor scanner is a novel instrument for rapid food detection developed in the 1990s. It uses specific sensors and pattern recognition systems to quickly provide overall information about the sample under test, indicating the underlying characteristics of the sample. It is an instrument composed of a selective array of electrochemical sensors and an appropriate identification method. It can identify simple and complex odors and can obtain results consistent with human sensory evaluation.
  • the discrimination index is a measurement index used by the electronic tongue and electronic nose to determine the overall discrimination effect of the sample, and its value range is [-100, +100]. -100,0 indicates that it cannot distinguish the taste and smell of the sample; +100 indicates that it can effectively distinguish, and the closer the value is to 100, the better the discrimination effect.
  • the plant functional salt obtained in the example was made into a 10% solution, and the difference in taste was detected with an electronic tongue (Smartongue, Isensogroup).
  • the electronic nose collects the odor signals of different samples through 14 sensors, and then analyzes the data by software for principal component analysis.
  • the plant functional salt obtained in the example is made into a 10% solution, and the electronic nose (iNose, Isensogroup) is used to detect the difference in taste.
  • the two principal components that account for the most information in the sample are abscissa PC1 and ordinate PC2, respectively.
  • the contribution rate of principal component 1 (PC1) is 99.80%, which is much larger than that of principal component 2. Therefore, the two sample data The farther the distance on the abscissa is, the larger the difference is.
  • mice Forty-five 5-week-old SPF C57BL / 6 healthy male mice were randomly divided into 9 groups after one week of adaptive feeding.
  • the experimental design and dosage configuration are shown in Table 9. Except for the normal control group (1 # ), which used conventional feed, the other 8 groups used salt-free basic feed.
  • Administration phase the oral dose results in mice according to the World Health Organization recommended daily salt intake of 5g per dose, and in standard animal equivalent conversion method, in addition to the group # 1, each sample was fed with 4% NaCl , Intragastric administration once a day for 21d.
  • the breeding environment temperature is (22 ⁇ 2) ° C, and the relative humidity is 50-70%.
  • model control group and the experimental group (# 3 - # 9) mice were injected with 0.1% CCl 4 in olive oil solution, CCl 4 at a dose of 10mL / kg.BW, normal control group (# 1 ⁇ 2 # ) Give an equal volume of olive oil and fast overnight.
  • mice Sixteen hours after the injection of the CCl 4 olive oil solution, the mice were sacrificed by spinal dislocation, the eyeballs were immediately removed for blood extraction, and the mice were dissected to take the liver.
  • the upper serum was collected for determination of blood biochemical indicators.
  • liver homogenate was centrifuged in an ultra-low temperature centrifuge (4000r / min, 20min), and the supernatant was taken to determine the content of malondialdehyde (MDA), glutathione (GSH), and total Activities of superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) (Coomassie blue method).
  • MDA malondialdehyde
  • GSH glutathione
  • T-SOD total Activities of superoxide dismutase
  • GSH-Px glutathione peroxidase
  • Liver tissue from the same part of each group of rats was taken, washed with cold saline, fixed with neutral formalin, 4 ⁇ m paraffin sections were made, and HE stained, and the morphological changes were observed under a microscope.
  • the experimental data were expressed as mean ⁇ standard error (x ⁇ s), and the data were tested for homogeneity of variance; the comparison of data between groups was performed by single factor analysis of variance.
  • the Duncan test in the one-way analysis of variance (ANOVA) of SPSS19.0 statistical software was used to analyze the significance, P ⁇ 0.05 was significant, and P ⁇ 0.01 was extremely significant.
  • mice in each experimental group increased with the feeding time, and the body weight did not show significant differences (P> 0.05). It was observed that the fur of the mice in each group was smooth and the clinical activities were normal. Good health status, indicating that intake of different salt samples had no effect on the clinical performance of mice. See Table 10 for the body weight of each group of experimental rats during the test cycle.
  • the positive drug control group (bifendate) had the strongest effect of inhibiting ALT activity, but had no effect on AST.
  • the commercially available Renshan Zizhu salt like the original salt, did not show an inhibitory effect on serum aminotransferase activity in mice with acute chemical liver injury.
  • # means P ⁇ 0.01;
  • # means P ⁇ 0.01;
  • Figure 14 is the effect of plant functional salts on the histopathology of the liver of experimental mice ( ⁇ 400).
  • # 1 and # 2 are mouse normal liver tissue, showing hepatic cords arranged in rows, clear cell borders, normal karyotype, no abnormalities; hepatic cords derangement 3 # NaCl model group karyotype change significantly, There is a large amount of cell infiltration in the central vein; hepatocyte cords in the 4 # biphenyldiester group are clearer, the karyotype is normal, and a small amount of vacuolar degeneration, indicating that bifendate drip pills have a therapeutic effect on acute liver injury; 5 # The arrangement of hepatocyte cords in Renshan Zizhu salt group was clear, the karyotype was normal, there was a small amount of vacuolar degeneration and watery degeneration; the hepatocyte cords of the # 6 original salt control group were irregularly arranged, the karyotype changed significantly, and a large number in
  • the edible salt of Chinese residents mainly comes from sea salt.
  • the sun-sea salt original salt
  • the results of this study show that the levels of GSH in the liver of experimental rats that ingested the raw salt were significantly lower than those in the same dose of pure NaCl, suggesting that there may indeed be some harmful factors in the liver that caused oxidative stress in the liver.
  • the final results show that the traditional ingredients of the bamboo salt roasted by the traditional method are limited in the effective components absorbed from the bamboo tube, and their liver-protective effects are not as good as the pine, bamboo, and plum salts provided by the present invention.
  • Tyrosine was used as a substrate for tyrosinase monophenolase activity measurement
  • L-DOPA was used as a substrate for tyrosinase diphenolase activity measurement
  • tyrosinase activity measurement was performed with reference to the method of Huang et al.
  • the raw salt, roasted salt, pine salt, bamboo salt, and plum salt were prepared into a salt solution of 0.2%, 1%, 5%, and 10% by mass with PBS solution, and the sample solutions of different concentrations and 0.1mol were accurately sucked in order according to the table.
  • Tyrosinase monophenolase / diphenolase inhibition rate /% [1- (A3-A4) / (A1-A2)] ⁇ 100
  • the MTT method was used to determine the proliferation rate of B16 cells.
  • the original salt was used as a negative control, and arbutin and Korean bamboo salt were used as positive controls.
  • the B16 cells in the logarithmic growth phase were selected and inserted into a 96-well plate. After the adherence, the original culture medium was discarded, and the prepared raw salt, pine salt, bamboo salt, containing 0.25%, 0.50%, 0.75%, and 1% by mass were added.
  • the culture medium of plum salt, -arbutin and Korean bamboo salt was cultured at 37 ° C and 5% CO 2 for 48 hours.
  • Dopa oxidation method was used to determine the tyrosinase activity in B16 cells.
  • B16 cells were inserted into a 96-well plate, the original medium was discarded after being adhered, and a medium containing each sample was added to culture for 48 h.
  • the culture solution of each group of cells that has reached the action time is decanted, washed twice with PBS buffer, and 50 ⁇ L of a 1% by mass Triton-X100 aqueous solution is added to each well, and then quickly stored in a -80 ° C refrigerator for 30 minutes and removed. After thawing at 37 ° C, the cells were completely ruptured. 10 ⁇ L of L-DOPA solution (10g / L) was added to each well.
  • Intracellular tyrosinase activity /% (OD 475 (experiment) -OD 475 (blank) ) / (OD 475 (control) -OD 475 (blank) ) / cell proliferation rate ⁇ 100
  • tyrosinase When tyrosine is used as a substrate, tyrosinase can catalyze it into dopa and further produce melanin. This process mainly exerts tyrosinase monophenolase activity.
  • the inhibitory effect of various salt solutions on tyrosinase monophenolase activity is shown in FIG. 15. As can be seen from FIG. 15, the original salt showed no significant inhibitory effect on tyrosinase in the concentration range of 0.2 to 10.0%. Roasted salt had the greatest effect on tyrosinase at a concentration of 5.0%, but the inhibition rate was not higher than 10%. Pine salt and bamboo salt have a significant effect on tyrosinase activity and show a concentration-effect relationship.
  • tyrosinase When dopa is used as a substrate, tyrosinase can catalyze it into dopaquinone and further produce melanin. This process mainly uses tyrosinase diphenolase activity.
  • the inhibition rate of tyrosinase diphenolase by various salt solutions is shown in FIG. 16. It can be seen from the figure that compared with the inhibition rate of tyrosinase monophenolase, the inhibition rate of tyrosinase diphenolase by the original salt and roasted salt is obviously enhanced, and there is a certain concentration-effect relationship, but the maximum inhibition rate does not exceed 30 %.
  • the tyrosinase diphenolase inhibition of pine salt and bamboo salt is similar to that of monophenolase.
  • the high concentration (5.0%, 10.0%) of the plum salt solution has a diphenolase inhibition rate that is the same as that of pine salt and bamboo salt.
  • the solution has little difference, and the inhibition rate can reach 92.0% at 5%.
  • plant salt has more potassium, iron, magnesium, calcium and other metal elements, which may have a certain effect on the protein structure of tyrosinase, thereby inhibiting its activity.
  • plant salt has no effect on B16 cells. Obvious extra cytotoxicity, and the cell proliferation rate in 0.5% pine salt, bamboo salt, and plum salt is much greater than the original salt, indicating that these plant salts have a certain protective effect on B16 cells in the NaCl environment.
  • the cell proliferation rate and concentration of the arbutin group in the positive control group had little relationship with each other, about 70%.
  • Tyrosinase is a key rate-limiting enzyme for melanin synthesis in B16 cells. Its enzyme activity is an important factor affecting melanin production and an important indicator for measuring the whitening efficacy of functional factors. It can be known from Table 15 that each sample has a certain effect on the tyrosinase activity of B16 cells, and the intracellular tyrosinase activity is significantly reduced, and there is a certain concentration-effect relationship. Compared with the original salt, pine salt, bamboo salt, and Korean bamboo salt have higher tyrosinase activity in each group of cells, indicating that these plant salts have less inhibitory effect on tyrosinase in B16 cells than the original salt.
  • the tyrosinase activity of the cells was lower than that of the original salt at the concentration, indicating that plum salt can significantly reduce the tyrosinase activity of B16 cells, and the effect is better than the test substances in each group.
  • the tyrosinase activity of cells in the positive control arbutin group was higher than that in the salt sample group of the same concentration.
  • tumor cells An important feature of tumor cells is that they can spread and metastasize during proliferation, and their ability to spread can be judged by their degree of migration.
  • the B16 cells in the normal group (a) basically healed after 48 h of culture, indicating that their migration ability is very strong.
  • B16 cells treated with the original salt (b) entered only part of the cells in the underlined area, indicating that the original salt had a certain inhibitory effect on cell migration.
  • Arbutin (c) also has a certain inhibitory effect on the mobility of B16 cells, but its effect is not as good as that of plant salts.
  • the cell cycle refers to the process from the end of cell division to the end of the next cell division. It is divided into four stages, G0 / G1 (pre-DNA synthesis), S (DNA replication), and G2 (DNA replication completed). To the beginning of mitosis), M phase (beginning to the end of cell division). The DNA content of cells in different cycles is different. Using the fluorescent dye PI to stain the cell DNA and detecting the fluorescence intensity in a flow cytometer, you can study the distribution of cells in different cycle stages and understand the cell proliferation.
  • MTT results show that each test substance has a certain inhibitory effect on the proliferation of the B16 cell population, and cell cycle studies have found that the viability of surviving cells is better. It is speculated that the treatment of the test substance may cause some cell death and be resistant to some The cells have the effect of promoting proliferation.
  • the pine, bamboo, and plum salts provided by the present invention exhibit concentration-dependent inhibition on monophenolase activity and diphenolase activity of tyrosinase in vitro.
  • the sample concentration is At 10%, the inhibition rates of tyrosinase monophenolase reached 97.8%, 94.2%, and 73.3%, respectively; at a concentration of 5%, the inhibition rates of diphenolase reached 99.9%, 93.0%, and 92.0%, respectively.
  • Plant functional salt has a certain inhibitory effect on B16 cell proliferation, but it has no obvious cytotoxicity compared to the original salt.
  • Plum salt can significantly reduce tyrosinase activity in B16 cells, and B16 cell tyrosinase activity in 0.75% plum salt medium is only 6.07%.
  • plant functional salts can significantly inhibit the lateral migration of B16 cells, but do not inhibit their proliferation by affecting the cell cycle.
  • mice male rats SD rats, 13-14 weeks old, weighing about 200g. After one week of adaptive feeding, they were randomly divided into 8 groups, namely the control group fed with normal salt, the control group fed with normal saline without salt, the high salt model group, the experimental group (functional salt of pine, bamboo, and plum plants), and the positive control group Felodipine sustained-release tablets), 4 rats in each group, and the experimental design and dosage configuration are shown in Table 17. During the experiment, the animals were raised in a single cage at a temperature of 25 ° C and a humidity of 30%. Clinical manifestations were observed weekly. During the administration period, the rats' daily high salt intake was 4000mg / kg (equivalent to 8% dietary salt).
  • the samples and control samples were configured to the corresponding concentration.
  • the solution was orally administered to rats daily with 2 mL / 100 g of bamboo saline, pine saline, and plum saline. Except for the normal control group (1 # ), which used conventional feed, the other 8 groups used salt-free basic feed.
  • Blood pressure and weight were measured every 4 days, and the test lasted 27 days. After the last gastric lavage, fasting and water were not allowed, placed in a metabolic cage for 24 hours and urine was collected. After the end, anesthetic was injected intraperitoneally, and the rats were sacrificed after blood was drawn from the heart.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • the high-salt test group 3 and the conventional raw salt group 2 had a significant difference, indicating that oral high-dose salt can lead to hypertension, and the experimental modeling was successful.
  • the functional salt groups of the pine, bamboo, and plum plants obtained in the examples all had significant differences from the original salt groups, indicating that ingestion of the plant functional salts prepared in this example had the effect of preventing hypertension.
  • plum salt has the most obvious antihypertensive effect in plant salt, and there is no significant difference with low salt intake. It may be due to the special physicochemical structure and element content of plum salt.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • ALT Alanine aminotransferase
  • ALT an enzyme involved in the metabolism of human proteins
  • ALT can accelerate the conversion of protein amino acids in the body, and it is the most abundant in liver cells.
  • tissues and organs are diseased, the ALT in them will be released into the blood, which will increase the serum ALT content.
  • ALT in the serum of the high-salt group was significantly increased, indicating that the liver was damaged.
  • AST Aspartate aminotransferase helps to understand the degree of damage to myocardium, liver and kidney tissue. AST exists in various tissues of the human body, with the heart muscle being the most abundant, followed by the liver. When necrosis of the heart and liver cells occurs, m-AST is released from the mitochondria, which increases the AST in the serum. The AST in the serum of rats in the high-salt group was significantly increased, indicating that the liver was affected to some extent after ingesting high concentrations of salt.
  • Alkaline phosphatase is widely distributed in human liver, bones, intestines, kidneys and placenta. It is excreted by the liver to the gallbladder. It is mainly used for the diagnosis and differential diagnosis of bone, hepatobiliary system diseases. Bone, liver and gallbladder diseases are closely related. There was no significant difference in ALP content between the groups, which indicated that the effect of high-salt diet on ALP in one month was not obvious.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • Urea nitrogen is a nitrogen-containing compound other than protein in the plasma. It is filtered from the glomerulus and excreted. When renal insufficiency is decompensated, BUN will increase. It is used clinically as an index for judging glomerular filtration function; creatinine (CRea) is a metabolic product of muscle tissue in the body, which is mainly excreted by glomerular filtration. When renal insufficiency, creatinine will accumulate in the body It becomes a harmful toxin to the human body; when all the uric acid in the blood is filtered from the glomeruli, abnormal urine processing by the kidneys will increase the blood uric acid (UA) content. It can be seen from the table that there is no significant difference between the indicators in each group within one month, and it may be that the high salt diet time is too short.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • CK creatine kinase
  • the measurement of creatine kinase activity can be used for the diagnosis of skeletal muscle disease and myocardial disease.
  • Pathological increase myocardial infarction, viral myocarditis, dermatomyositis, muscular dystrophy, pericarditis, accidental rupture of cerebral blood vessels, etc. It can be seen from the table that high salt concentration has a significant effect on CK content, and a high salt diet for more than a month may have a certain effect on myocardial function.
  • the CK content of pine salt is significantly lower than that of the high salt group, indicating that pine salt has the potential to prevent myocardial damage.
  • Lactate dehydrogenase is found in almost all tissues and is most abundant in heart, skeletal muscle, and kidney, and can be used for the diagnosis of myocardial disease. Increased lactate dehydrogenase: mainly seen in myocardial infarction, hepatitis, malignant tumors, pulmonary infarction, leukemia, hemolytic anemia, kidney disease, progressive muscle atrophy, etc. The contents of CK and LDH in the high-salt diet were lower than those in the normal salt diet group, which further explained that the high-salt diet had damage to myocardial function.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • Urinary creatinine is mainly from the blood. After filtering through the glomerulus, it is eliminated with the urine. The renal tubules are basically not absorbed and excreted rarely. Urine creatinine content changes when kidney problems occur. Normal urine contains a small amount of small molecule protein (U-TP). When protein in the urine increases, proteinuria is formed. Proteinuria is a common manifestation of kidney disease, and proteinuria can also occur in systemic diseases. Urinary microalbumin (U-ALB) is one of the important plasma proteins. Under normal circumstances, albumin has a large molecular weight and cannot cross the glomerular basement membrane.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • each of the high-salt test groups (groups 4 to 8) * indicates p ⁇ 0.05, and ** indicates p ⁇ 0.01.
  • the systolic blood pressure of male SD rats significantly increased after taking raw salt, and showed obvious liver, kidney, and myocardial function damage; while the blood pressure of experimental animals ingesting the plant functional salt of the present invention was significantly lower than the original salt , Indicating that it has the potential to replace ordinary edible salt and prevent hypertension.

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Abstract

Disclosed are a functional plant salt and a preparation method therefor. The preparation method comprises mixing one or more plant extract powders, in an addition amount of 1%-40%, with a fine salt until uniform, baking same all once or in batches at a high temperature of 800℃-1400℃, so as to form functional plant salts with different colors, different space structures, different physicochemical properties, different flavors and different uses.

Description

新型竹盐及其制备方法Novel bamboo salt and preparation method thereof 技术领域Technical field
本发明涉及食品和保健食品技术领域，具体涉及一种功能性植物盐及其制备方法，更具体地涉及一种竹盐及其制备方法。The invention relates to the technical field of food and health food, in particular to a functional plant salt and a preparation method thereof, and more particularly to a bamboo salt and a preparation method thereof.
背景技术Background technique
植物盐是指从植物有机体进行提取、用纯粹物理流程和工艺、符合低钠盐和多种矿物质微量元素群、各种元素比例符合人体要求的一种平衡盐类，对人体的微量元素平衡、机能调节和新陈代谢及健康具有一定促进作用的新型食用盐。Plant salt refers to a kind of balanced salt extracted from plant organisms, using pure physical processes and processes, conforming to low sodium salts and a variety of mineral trace element groups, and various element proportions meeting the requirements of the human body. , A new type of edible salt with a certain promoting effect on function regulation and metabolism and health.
植物盐的主要特点：①以植物为主要来源；②天然低钠盐和天然平衡盐；③富含多种有机营养物质和生物活性成分；④加工生产过程中不添加任何化学合成物质；⑤对矿物元素群缺乏或不平衡导致的多种疾病及相关并发症具有一定辅助治疗和调节作用。如对钠敏感性高血压、2型糖尿病、高血脂、心血管疾病、肿瘤和机体免疫机能障碍等疾病和并发症等疾病有明确的疗效。The main characteristics of plant salt: ① Plants are the main source; ② Natural low sodium salt and natural balanced salt; ③ Rich in various organic nutrients and biologically active ingredients; ④ No chemical synthetic substances are added during processing and production; ⑤ A variety of diseases and related complications caused by the lack or imbalance of mineral element groups have certain adjuvant treatment and regulating effects. Such as sodium-sensitive hypertension, type 2 diabetes, hyperlipidemia, cardiovascular disease, tumors and immune dysfunction and other diseases and complications have a clear effect.
植物盐(Plant-Salt)概念由香港预防医学研究院院长潘汉杰教授首次提出，并不断进行完善和发展。The concept of plant salt (Plant-Salt) was first proposed by Professor Pan Hanjie, Dean of the Hong Kong Institute of Preventive Medicine, and it has been continuously improved and developed.
潘汉杰教授指出，植物盐的功效同原料植物的选种、培植土壤和浇灌水源的活性等具有非常密切的关系。植物盐工程在我国刚刚起步，前景十分广阔。Professor Pan Hanjie pointed out that the efficacy of plant salt is closely related to the selection of raw materials, the activity of cultivating soil and watering sources. The plant salt project has just begun in China and has a very bright future.
潘汉杰教授强调，植物盐的生物学活性，取决于原料植物品种的选择和品种组合以及它们所含无机盐种类的多少、元素的含量和比例关系及植物相关的生物学活性成分等因素；植物盐主要针对需要低钠饮食人群，和因机体微量元素不平衡或缺乏导致机体机能代谢障碍、机能下降和功能受损的人群。Professor Pan Hanjie emphasized that the biological activity of plant salts depends on the selection of plant species and the combination of varieties of raw materials, the number of types of inorganic salts they contain, the content and proportion of elements, and the biologically active ingredients related to plants; plant salts It is mainly aimed at people who need a low-sodium diet, and people who have a disordered or impaired function due to imbalance or lack of trace elements in the body.
发明内容Summary of the Invention
本发明要解决的技术问题是提供一种功能性植物盐，及该功能性植物盐的制备方法。The technical problem to be solved by the present invention is to provide a functional plant salt and a method for preparing the functional plant salt.
在本发明中，所述功能性植物盐为竹盐，是指以竹提取物为植物性原料所制得的产物。In the present invention, the functional plant salt is bamboo salt, which refers to a product prepared by using bamboo extract as a plant-based raw material.
本发明的第一方面，提供了一种功能性植物盐，所述功能性植物盐是如下制备的：将一种或多种植物提取物粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温下一次或分次烤制，获得功能性植物盐。According to a first aspect of the present invention, a functional plant salt is provided. The functional plant salt is prepared by mixing one or more plant extract powders with fine salt in an amount of 1% to 40%. Uniform, roasted once or in multiples at a high temperature of 800-1400 ° C to obtain a functional plant salt.
在另一优选例中，所述植物提取物为竹提取物。In another preferred example, the plant extract is a bamboo extract.
在另一优选例中，所述植物提取物选自下组：竹叶提取物、竹茹提取物、竹竿提取物、竹笋提取物、或其组合。In another preferred example, the plant extract is selected from the group consisting of bamboo leaf extract, bamboo rutberry extract, bamboo pole extract, bamboo shoot extract, or a combination thereof.
在另一优选例中，所述植物提取物选自下组：竹叶黄酮、竹叶多酚、或其组合。In another preferred example, the plant extract is selected from the group consisting of bamboo leaf flavones, bamboo leaf polyphenols, or a combination thereof.
在另一优选例中，所述细盐选自下组：海盐、湖盐、井盐、矿盐。In another preferred example, the fine salt is selected from the group consisting of sea salt, lake salt, well salt, and mineral salt.
在另一优选例中，所述细盐为粗制海盐。In another preferred example, the fine salt is crude sea salt.
在另一优选例中，所述功能性植物盐为竹盐。In another preferred example, the functional plant salt is bamboo salt.
在另一优选例中，所述植物提取物还包含梅提取物和/或松提取物。In another preferred example, the plant extract further comprises a plum extract and / or a pine extract.
在另一优选例中，所述细盐为食盐。In another preferred example, the fine salt is common salt.
在另一优选例中，所述功能性植物盐的ΔL值为28-50，较佳地35-50。In another preferred example, the ΔL value of the functional plant salt is 28-50, preferably 35-50.
在另一优选例中，所述功能性植物盐的ΔL值为30-40，较佳地32-39。In another preferred example, the ΔL value of the functional plant salt is 30-40, preferably 32-39.
本发明的第二方面，提供了一种本发明第一方面所述的功能性植物盐的制备方法，包括步骤：将一种或多种植物提取物粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温下一次或分次烤制，获得功能性植物盐。According to a second aspect of the present invention, there is provided a method for preparing a functional plant salt according to the first aspect of the present invention, comprising the steps of: adding one or more plant extract powders in an amount of 1% to 40% and Fine salt is mixed uniformly, and it is roasted once or in multiples at a high temperature of 800-1400 ° C to obtain a functional plant salt.
在另一优选例中，所述烤制在机械化烤炉中进行。In another preferred example, the roasting is performed in a mechanized oven.
在另一优选例中，所述烤制在中频熔炼炉中进行。In another preferred example, the roasting is performed in an intermediate frequency melting furnace.
本发明的第三方面，提供了一种食品或药品组合物，包含一种或多种本发明第一方面所述的功能性植物盐。According to a third aspect of the present invention, there is provided a food or pharmaceutical composition comprising one or more functional plant salts according to the first aspect of the present invention.
本发明的第四方面，提供了一种本发明第一方面所述的功能性植物盐的用途，用于制备选自下组的物质：药物助剂、保健食品/饮品、功能性食品/饮品、餐桌调味盐、烹饪盐、特殊用途化妆品或个人护理用品。According to a fourth aspect of the present invention, there is provided the use of the functional plant salt according to the first aspect of the present invention, for preparing a substance selected from the group consisting of a pharmaceutical auxiliary, a health food / drink, and a functional food / drink , Table seasoning salt, cooking salt, special purpose cosmetics or personal care products.
本发明的第五方面，提供了一种抑制转氨酶活性的方法，包括步骤：将一种或多种抑制有效量的本发明第一方面所述的功能性植物盐施用于所需患者。According to a fifth aspect of the present invention, there is provided a method for inhibiting transaminase activity, comprising the step of applying one or more inhibitory effective amounts of the functional plant salt according to the first aspect of the present invention to a desired patient.
在另一优选例中，所述转氨酶选自下组：天冬氨酸转氨酶(ALT)、谷氨酸转氨酶(AST)。In another preferred example, the aminotransferase is selected from the group consisting of aspartate aminotransferase (ALT) and glutamic acid aminotransferase (AST).
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。It should be understood that, within the scope of the present invention, the above technical features of the present invention and the technical features specifically described in the following (such as the embodiments) may be combined with each other to form a new or preferred technical solution. Due to space limitations, I will not repeat them here.
附图说明BRIEF DESCRIPTION OF THE DRAWINGS
图1是实施例4所得松盐的照片。FIG. 1 is a photograph of the pine salt obtained in Example 4. FIG.
图2是实施例5所得竹盐的照片。FIG. 2 is a photograph of bamboo salt obtained in Example 5. FIG.
图3是实施例6所得梅盐的照片。FIG. 3 is a photograph of plum salt obtained in Example 6. FIG.
图4是实施例7所得松盐的照片。FIG. 4 is a photograph of the pine salt obtained in Example 7. FIG.
图5是实施例8所得竹盐的照片。FIG. 5 is a photograph of bamboo salt obtained in Example 8. FIG.
图6是实施例9所得梅盐的照片。FIG. 6 is a photograph of plum salt obtained in Example 9. FIG.
图7是不同配比和不同工艺参数下获得的植物盐照片。FIG. 7 is a photograph of plant salts obtained under different formulations and different process parameters.
图8是不同盐的显微图片。Figure 8 is a micrograph of different salts.
图9是不同盐的SEM结果。Figure 9 is the SEM results of different salts.
图10是不同盐的TEM结果。Figure 10 is the TEM results of different salts.
图11是不同盐的XRD结果。Figure 11 is the XRD results of different salts.
图12是不同盐的电子舌测试结果。Figure 12 shows the results of the electronic tongue test for different salts.
图13是不同盐的电子鼻测试结果。Figure 13 shows the results of electronic nose tests for different salts.
图14是各组小鼠肝组织病理切片的形态学观察结果。FIG. 14 is a morphological observation result of pathological sections of liver tissue of mice in each group.
图15是不同浓度的盐溶液对体外酪氨酸酶(单酚酶)的抑制作用，**表示与原盐比较，实验组存在极显著差异(p＜0.01)，n＝3。Figure 15 shows the inhibitory effect of different concentrations of salt solution on tyrosinase (monophenolase) in vitro. ** indicates that compared with the original salt, there is a very significant difference in the experimental group (p <0.01), n = 3.
图16是不同浓度盐溶液对体外酪氨酸酶(二酚酶)的抑制作用，**表示与原盐比较，实验组存在极显著差异(p＜0.01)，n＝3。Figure 16 shows the inhibitory effect of different concentrations of salt solution on tyrosinase (diphenolase) in vitro. ** indicates that compared with the original salt, there is a very significant difference in the experimental group (p <0.01), n = 3.
图17为0.5％质量分数的各试样对B16细胞迁移的影响，a—正常；b—原盐；c—熊果苷；d—松盐；e—竹盐；f—梅盐。Figure 17 is the effect of 0.5% mass fraction of each sample on the migration of B16 cells, a-normal; b-original salt; c-arbutin; d-pine salt; e-bamboo salt; f-plum salt.
图18为0.5％质量分数的各试样对B16细胞周期的影响，a—正常；b—原盐；c—熊果苷；d—松盐；e—竹盐；f—梅盐。Figure 18 shows the effect of 0.5% mass fraction of each sample on the B16 cell cycle, a-normal; b-raw salt; c-arbutin; d-pine salt; e-bamboo salt; f-plum salt.
图19为6.3.1中实验动物的体重变化图。FIG. 19 is a graph showing changes in body weight of experimental animals in 6.3.1.
图20为6.3.2中实验大鼠的血压变化。Figure 20 shows the changes in blood pressure in experimental rats in 6.3.2.
具体实施方式detailed description
本发明人经过长期而深入的研究，通过采用未经预烤的竹提取物与原盐的混合物在大于原盐熔点以上的温度(如800-1400℃)进行高温处理，意外地制备了一种具有优异的生物学效应的竹盐。所述竹盐的制备方法简单、易于大规模推广。在此基础上，发明人完成了本发明。After a long and intensive study, the inventors have unexpectedly prepared a kind of high-quality treatment by using a mixture of unpre-roasted bamboo extract and raw salt at a temperature higher than the melting point of the raw salt (such as 800-1400 ° C). Biological effects of bamboo salt. The preparation method of the bamboo salt is simple and easy to popularize on a large scale. On this basis, the inventors have completed the present invention.
术语the term
在本发明中，术语“功能性植物盐”为竹盐，是指以竹提取物为植物性原料所制得的产物。In the present invention, the term "functional plant salt" is bamboo salt, which refers to a product prepared by using bamboo extract as a plant-based raw material.
在本发明中，术语“未经预烤”是指制备所述竹盐的原料为竹提取物与原盐的直接混合物，该混合物未经传统制备竹盐的一次或多次低温(如800℃以下)预烘烤处理。In the present invention, the term "without pre-baking" means that the raw material for preparing the bamboo salt is a direct mixture of bamboo extract and raw salt, and the mixture has not been subjected to one or more low temperatures (such as below 800 ° C) of traditional preparation of bamboo salt. ) Pre-baking treatment.
在本发明中，术语“植物提取物”为竹提取物。In the present invention, the term "plant extract" is a bamboo extract.
竹提取物Bamboo extract
所述竹提取物选自下组：竹叶提取物、竹茹提取物、竹竿提取物、竹笋提取物、或其组合。The bamboo extract is selected from the group consisting of bamboo leaf extract, bamboo rut extract, bamboo pole extract, bamboo shoot extract, or a combination thereof.
在另一优选例中，所述竹提取物选自下组：竹叶黄酮、竹叶多酚、竹茹三萜、竹多糖(如竹茹多糖)、竹笋甾醇、竹笋氨基酸肽、或其组合。In another preferred example, the bamboo extract is selected from the group consisting of bamboo leaf flavones, bamboo leaf polyphenols, bamboo triterpenes, bamboo polysaccharides (such as bamboo polysaccharides), bamboo shoot sterols, bamboo shoot amino acid peptides, or combinations thereof .
竹盐及其制备方法和用途Bamboo salt and preparation method and application thereof
目前广为人知的韩国竹盐实际起源于中国。相传1300多年前，在茶马古道上有一支商队借宿于寺庙，晚上马厩起火，将白色的食盐与装盐的容器(竹篓)一起烧成了灰色的盐块(即竹盐的雏形)，僧侣们将其用于治疗百姓的消化不良、外伤止血和消毒等效果显著，渐渐流传开来。迄今，在云南的一些寺庙、浙江某些山区，仍有人将食盐装在竹筒中放入灶火中煅烧后取食。《本草纲目》中也有将食盐放入瓷器皿中、炒制后药用的记载。民间应用历史及现代科学研究表明，竹盐具有一定的抗炎、抗过敏、抗癌、抑菌、预防肥胖、保护牙齿、抗胃溃疡、护肝、抗衰、预防高血压、防治血液酸化、改善糖尿病、治疗关节炎、增强免疫力等功效。The well-known Korean bamboo salt actually originated in China. According to legend, more than 1300 years ago, a caravan stayed at a temple on the Ancient Tea Horse Road. At night, the horse stables caught fire, and the white table salt and the container (bamboo basket) containing salt were burned into gray salt blocks (that is, the prototype of bamboo salt). The monks used it to treat people's indigestion, trauma, hemostasis, and disinfection. The effects were significant and gradually spread. So far, in some temples in Yunnan and some mountainous areas in Zhejiang province, some people still put salt in bamboo tubes and calcined them in the stove to eat. "Compendium of Materia Medica" also records the use of table salt in porcelain ware, and medicated after being fried. Folk application history and modern scientific research show that bamboo salt has certain anti-inflammatory, anti-allergic, anti-cancer, antibacterial, prevent obesity, protect teeth, anti-gastric ulcer, liver protection, anti-aging, prevent hypertension, prevent blood acidification, Improve diabetes, treat arthritis, and enhance immunity.
传统的竹盐生产是以日晒海盐为原料，将其装灌于新鲜的楠竹(即毛竹)筒中，用黄土封口，以松木为燃料，在土窑中反复烤制而成，根据充填、烤制的次数多寡区分品质与等级。九烤竹盐即需要重复“充填—烤制—冷却—粉碎—再充填”的9次循环，目的是为了让盐更多地吸取新鲜竹筒中的有益成分；并在最后一次烤制时，在松木中添加松脂，使烤制温度达到1300℃，冷却后得到蓝紫色的盐，称为紫竹盐。一个生产周期大约需要一个月。Traditional bamboo salt production uses sun-sea salt as raw material, which is filled into fresh bamboo bamboo (that is, moso bamboo), sealed with loess, pine wood as fuel, and repeatedly baked in an earth kiln. According to the filling, The number of roasts distinguishes between quality and grade. Nine roasted bamboo salt needs to repeat 9 cycles of "filling-roasting-cooling-pulverizing-refilling", the purpose is to let the salt absorb more beneficial ingredients from the fresh bamboo tube; and during the last roasting, Pine resin is added to pine wood to make the baking temperature reach 1300 ° C. After cooling, blue-violet salt is obtained, which is called purple bamboo salt. A production cycle takes about one month.
国内有名的竹盐生产厂家(浙江临安三和园)将传统烤制过程描述如下：100～300℃下脱去水分，300～800℃时原盐和竹筒中的有机质燃烧殆尽；800～1200℃时原盐中的部分重金属元素以氯化物的形式被蒸馏除去，同时竹筒和黄泥中的微量元素与氯化钠结合；在1300℃以上的高温下将熔融态的食盐取出，快速冷却导致1晶格发生畸变，即可得到竹盐。A well-known domestic bamboo salt manufacturer (Sanhe Garden, Lin'an, Zhejiang) described the traditional roasting process as follows: remove moisture at 100-300 ° C, and burn the raw salt and organic matter in the bamboo tube at 300-800 ° C; 800-1200 ° C Some of the heavy metal elements in the time salt are distilled away in the form of chloride, and the trace elements in bamboo tube and yellow mud are combined with sodium chloride; the molten table salt is taken out at a high temperature above 1300 ° C, and rapid cooling causes 1 crystal. Distortion of the grid can get bamboo salt.
NaCl的熔点为801℃，要使得食盐完成融熔的温度至少要达到900℃左右。目前市面上真正意义上的人造植物盐只有竹盐一种，这是因为竹筒能作为盛盐的容器在高温下一同烤制，当竹筒碳化的同时，其所含的有益成分融进了盐中。然 而这种方式能吸收到的竹子有效成分十分有限，故需要反复循环，如九烤竹盐(紫竹盐)就需要循环九次。同时，还由于传统以松木为燃料的炉窑一般能达到的温度在800℃左右，为了使盐最终能够融熔，第九次烤制时采用在松枝中添加松脂的方法，以提高炉温，声称能达到1300℃以上(是缺乏实测数据支撑的，估计实际能达到的温度在900℃左右)。The melting point of NaCl is 801 ° C. The temperature for melting the salt should reach at least about 900 ° C. At present, there is only one kind of artificial plant salt in the real sense on the market. This is because the bamboo tube can be roasted together at a high temperature as a container for salt. When the bamboo tube is carbonized, the beneficial ingredients contained in it are melted into the salt. . However, the effective ingredients of bamboo that can be absorbed in this way are very limited, so it needs to be recycled repeatedly. For example, nine roasted bamboo salt (purple bamboo salt) needs to be recycled nine times. At the same time, because the traditional pine wood-fired kiln can generally reach a temperature of about 800 ° C, in order to finally melt the salt, the method of adding turpentine to pine branches during the ninth roasting to increase the furnace temperature, Claimed to be able to reach more than 1300 ° C (the lack of support of measured data, it is estimated that the actual temperature can reach about 900 ° C).
传统竹盐生产的主要瓶颈：一是生产周期长，劳动强度大，生产效率低；二是机械化程度低，产能十分有限，生产成本高；三是难以实现标准化和清洁化生产，由此导致的最大问题是难以实现大规模的工业化生产。迄今为止，竹盐始终是个小众商品，无法惠及更多的消费者。The main bottlenecks of traditional bamboo salt production are: one is the long production cycle, high labor intensity, and low production efficiency; the second is the low degree of mechanization, the production capacity is very limited, and the production cost is high; The biggest problem is that it is difficult to achieve large-scale industrial production. So far, bamboo salt has been a niche product and cannot reach more consumers.
本发明公开了一类功能性植物盐及其制备方法和用途，涉及食品、调味品、日用化学品和药品等领域，目的是提供一种天然、安全、营养、并具有多重生物学保健功效的植物盐。其核心是采用中频熔炼技术或电阻加热技术，将不同种类、不同来源的植物提取物与食用盐按一定质量比(0～40％)混合均匀后放入中频熔炼炉或电阻炉中，在常压下升温至800～1400℃融熔，冷却后结晶，从而得到具有不同色泽、不同理化性质、不同晶格结构、不同风味以及不同用途的植物盐系列。其显著特征是，盐溶液由弱酸性变为强碱性，色泽由白色变为灰色、浅灰色、淡粉色、紫红色和亮蓝色等不同颜色，呈现晶莹剔透的干净外观，并伴有淡淡的硫味；盐的晶胞尺寸较原盐更小，元素组成更为丰富，其中钾含量提高约100倍，钙约7倍，其他有益元素也显著增加。功能性植物盐的健康功效包括但不限于：美白、护肝、健胃、解毒、抗炎、抗过敏、清洁口腔、预防高血压、治疗偏头痛、抑制肠道癌变，可广泛应用于食品、饮品、调味品、保健食品、药品、日用化学品、个人护理用品等，前景十分广阔。由于氯化钠晶格的熔点为801℃，要使得食盐完全融熔的温度至少在900℃左右，而传统的炉窑一般无法满足该温度要求。中频熔炼和电阻加热是两种能将电能高效转化为热能的技术，可快速到达900～1700℃之间的熔炼温度，生产效率高，且可实现机械化、自动化、规模化、标准化和清洁化生产。同时，可通过控制植物提取物的种类、添加比例、熔炼温度、保温时间、以及升降温速率得到不同颜色、性能稳定、品质优良、质量可标化的功能性植物盐系列。The invention discloses a class of functional plant salts, a preparation method and uses thereof, and relates to the fields of food, condiments, daily chemicals, and medicines, and the purpose is to provide a natural, safe, nutritious, and multiple biological health care effect. Plant salt. The core is to use intermediate frequency melting technology or resistance heating technology. Mix different types of plant extracts from different sources with edible salt in a certain mass ratio (0-40%) and put them into an intermediate frequency melting furnace or resistance furnace. The temperature is reduced to 800 to 1400 ° C. and melted, and then crystallized after cooling, so as to obtain plant salt series with different colors, different physical and chemical properties, different lattice structures, different flavors, and different uses. Its distinctive feature is that the salt solution changes from weak acidity to strong alkalinity, and the color changes from white to gray, light gray, pale pink, fuchsia, and bright blue and other colors, showing a clean and clear appearance with a touch of light The sulphur taste of the salt is smaller than that of the original salt, and the elemental composition is more abundant. Among them, the potassium content is increased by about 100 times, the calcium is about 7 times, and other beneficial elements are also significantly increased. The health benefits of functional plant salts include, but are not limited to: whitening, liver protection, stomach strengthening, detoxification, anti-inflammatory, anti-allergy, oral cleansing, prevention of hypertension, treatment of migraine, inhibition of intestinal canceration, and can be widely used in food, Beverages, condiments, health foods, medicines, daily chemicals, personal care products, etc. have a bright future. Since the melting point of the sodium chloride crystal lattice is 801 ° C, the temperature for completely melting the table salt is at least about 900 ° C, and the traditional furnace cannot generally meet this temperature requirement. Intermediate frequency melting and resistance heating are two technologies that can efficiently convert electrical energy into thermal energy, which can quickly reach melting temperatures between 900 and 1700 ° C, have high production efficiency, and can achieve mechanization, automation, scale, standardization and clean production . At the same time, functional plant salt series with different colors, stable performance, excellent quality, and standardizable quality can be obtained by controlling the type of plant extracts, the addition ratio, the smelting temperature, the holding time, and the heating and cooling rates.
本发明创造性地应用中频熔炼技术或电阻加热技术，将食盐与不同的植物提取物混合后，在800～1400℃的高温下进行熔炼，形成了具有不同色泽、不同晶格结构、不同理化指标、不同风味、不同用途的功能性植物盐系列。The invention creatively applies intermediate frequency melting technology or resistance heating technology, and after mixing table salt with different plant extracts, it is smelted at a high temperature of 800 to 1400 ° C to form different colors, different lattice structures, different physical and chemical indexes, Functional plant salt series with different flavors and uses.
中频熔炼是一种常用的金属炼制手段，即将工频50Hz的交流电转变为中频(300～1000Hz)，把三相工频交流电整流后变成直流电，再把直流电变为可调节的中频电流，供给由电容和感应线圈里流过的中频交变电流，在感应线圈中产生高密度的磁力线，通过导磁的容器(如石墨碳化硅坩埚)迅速生产高温，以此加热物料(即盐与提取物的混合物)使其熔融。中频熔炼技术可以在短时间内达 到所需的熔炼温度，大幅度提高生产效率。Intermediate frequency smelting is a commonly used metal smelting method, that is, to convert AC power at 50Hz to intermediate frequency (300 ~ 1000Hz), rectify the three-phase power frequency AC to DC, and then convert DC to adjustable intermediate frequency current. Supply the intermediate frequency alternating current flowing through the capacitor and the induction coil to generate high-density magnetic lines of force in the induction coil, and quickly produce high temperature through a magnetically permeable container (such as a graphite silicon carbide crucible) to heat the material (i.e. salt and extraction Mixture of substances) to make it melt. Intermediate frequency melting technology can reach the required melting temperature in a short time, greatly improving production efficiency.
电阻加热技术是通过电阻体将电能转化为热能，从而对物料进行快速加热。我国目前商用的电阻炉主要以钼棒、硅棒或碳棒为加热元件，最高炉温可达1700℃。电阻加热技术可控制加热的时间、温度、升温速率等，从而做到工艺参数的精准控制和产品的标准化。The resistance heating technology converts electrical energy into heat energy through a resistor body, thereby rapidly heating the material. The resistance furnaces currently in commercial use in China mainly use molybdenum rods, silicon rods or carbon rods as heating elements, and the maximum furnace temperature can reach 1700 ° C. Resistance heating technology can control heating time, temperature, heating rate, etc., so as to achieve precise control of process parameters and standardization of products.
植物提取物又称植物化学素(即植物次生代谢产物)，是指采用不同技术手段从各种水果、蔬菜、花卉、中草药、药食两用植物、新资源食品中提取、分离、纯化得到的不同精度制剂乃至特征性的单体化合物，如来自于梅、兰、竹、菊、松、荷、茶、石榴、海棠、食用菌、藻类等的提取物。Plant extracts, also known as phytochemicals (that is, plant secondary metabolites), refer to the use of different technical means to extract, isolate, and purify from various fruits, vegetables, flowers, Chinese herbal medicine, medicinal and edible plants, and new resource foods. Different precision formulations and even characteristic monomer compounds, such as extracts from plum, orchid, bamboo, chrysanthemum, pine, lotus, tea, pomegranate, begonia, edible fungi, algae and the like.
藻类提取物的原料可以是海藻(如海带、裙带菜、羊栖菜等)，也可以是淡水或海淡水养殖的藻类，如蓝藻、绿藻、小球藻等等；食用菌提取物包括香菇、蘑菇、金针菇、竹荪、口蘑、木耳、银耳、猴头、灵芝、松露、牛肝菌等及其加工剩余物(如香菇柄等)的提取物；微生物来源的抽提物包括各类酵母的抽提物(又称酵母膏)，如啤酒酵母、面包酵母等。The raw material of the algae extract can be seaweed (such as kelp, wakame, sargassum, etc.), or algae cultivated in freshwater or seawater, such as cyanobacteria, green algae, chlorella, etc .; edible mushroom extracts include shiitake mushrooms , Mushrooms, enoki mushrooms, bamboo mushrooms, mushrooms, fungus, white fungus, monkey head, ganoderma, truffle, boletus, etc. and their processing residues (such as mushroom handles, etc.); microbial-derived extracts include various types of yeast Extracts (also known as yeast extract), such as beer yeast, baker's yeast, and the like.
本发明的主要技术特征为：采用中频熔炼技术或电阻加热技术，将不同来源的植物提取物与食盐按0～40％的比例混匀后(提取物粉末将均匀地吸附在食盐颗粒的表面)，在常压下升温至800～1400℃共同熔炼，冷却后重新结晶，获得了具有不同色泽、不同晶格结构、不同理化指标、不同风味和不同用途的植物功能盐系列。根据植物提取物的来源及其有效组分、添加比例、熔炼参数等的不同，得到的新型植物盐系列具有美白、护肝、健胃、解毒、抗炎、抗过敏、清洁口腔、预防高血压、治疗偏头痛、抑制肠道癌变等多重功能，可广泛应用于食品、饮品、调味品、保健食品、药品、日用化学品、个人护理用品等领域。The main technical features of the present invention are: after using intermediate frequency melting technology or resistance heating technology, plant extracts from different sources are mixed with table salt at a ratio of 0 to 40% (the extract powder will be evenly adsorbed on the surface of the table salt particles) , Under normal pressure, the temperature was raised to 800 ～ 1400 ℃, and then smelted. After cooling, it was recrystallized to obtain plant functional salt series with different colors, different lattice structures, different physical and chemical indexes, different flavors and different uses. According to the source of plant extracts and their effective components, addition ratios, melting parameters, etc., the new plant salt series has whitening, liver protection, stomach strengthening, detoxification, anti-inflammatory, anti-allergy, oral cleansing, and prevention of hypertension , Multiple functions such as treating migraine and inhibiting intestinal cancer, can be widely used in food, beverages, condiments, health foods, pharmaceuticals, daily chemicals, personal care products and other fields.
本发明大致的工艺过程和工艺参数如下：The approximate process and process parameters of the present invention are as follows:
按0～40％的质量比(优选5～25％)，将植物提取物粉末与食盐均匀混合；Mix the plant extract powder with table salt at a mass ratio of 0 to 40% (preferably 5 to 25%);
将以上混合物料放入导磁坩埚内置于中频熔炼炉中，开始升温，升温速率为1～200℃/min；或将以上混合物料放入刚玉坩埚内置于电阻炉中，开始升温，升温速率为1～20℃/min；Put the above mixture into the magnetically-conducting crucible and build it in the intermediate frequency melting furnace, and start to heat up, and the heating rate is 1 ～ 200 ℃ / min; 1 ～ 20 ℃ / min;
在常压下升至800～1400℃之间的某一温度保温0～6h；Rise to a temperature between 800 ～ 1400 ℃ under normal pressure for 0 ～ 6h;
将物料倒出，自然降温或控制性降温；Pour out the material and cool it naturally or in a controlled way;
冷却结晶后，粉碎、过筛得到产品；After cooling and crystallization, crushing and sieving to obtain the product;
加热温度根据需要可设置在800～1400℃之间的1个或多个区间内；The heating temperature can be set in one or more intervals between 800 and 1400 ° C as required;
以上混合—熔融—重结晶—冷却—粉碎的过程可一次或多次进行。The above mixing-melting-recrystallization-cooling-pulverizing process can be performed one or more times.
(1)本发明所涉及的原料食盐(又称原盐)，可以是海盐、湖盐、井盐、矿盐，优选粗制的海盐(即日晒盐)。(1) The raw common salt (also called raw salt) according to the present invention may be sea salt, lake salt, well salt, mineral salt, and preferably crude sea salt (that is, sun salt).
(2)本发明提供的功能性植物盐，根据植物提取物来源、添加比例、熔炼温度等参数的不同，其健康效应可具体表现为，非竹类的功能 性植物盐(如松盐、梅盐)的健康功效包括但不限于：美白、护肝、预防高血压。其应用领域包括食品、饮品、调味品、保健食品、药品、日用化学品和个人护理用品等。(2) According to the functional plant salt provided by the present invention, according to different sources of plant extract, addition ratio, melting temperature and other parameters, its health effect can be specifically expressed as non-bamboo functional plant salt (such as pine salt, plum, etc.) The health benefits of salt) include, but are not limited to, whitening, liver protection, and prevention of hypertension. Its applications include food, beverages, condiments, health foods, pharmaceuticals, household chemicals and personal care products.
(3)此外，本发明的中频熔炼技术和电阻加热技术也可直接用于加工食盐,即在制造过程中不添加任何植物提取物或其他外来物质，将原盐转化为食用安全性更高、外观更加晶莹剔透的烤盐。(3) In addition, the intermediate frequency smelting technology and the resistance heating technology of the present invention can also be directly used for processing table salt, that is, without adding any plant extract or other foreign substances in the manufacturing process, the raw salt is converted into a food with higher safety and appearance More crystal clear grilled salt.
本发明的突出优势在于，采用中频熔炼技术或电阻加热技术，可将原盐与任何来源、任何比例的植物提取物(粉末)混合，在精准设定的温度区间内共融后重结晶，通过一次或多次烤制得到具有特殊生物学功效的盐品，并可实现机械化、自动化、标准化、清洁化的大规模工业化生产。本发明创制出了以松盐和梅盐为代表的非竹系列植物功能盐，这是对食盐产业(尤其是中高端盐品市场)的一次革命，对全人类的健康事业具有十分重要的意义。The outstanding advantage of the present invention is that by using intermediate frequency melting technology or resistance heating technology, raw salts can be mixed with plant extracts (powder) of any source and ratio, and recrystallized after co-melting in a precisely set temperature interval, and once through Or it can be roasted several times to obtain salt with special biological effects, and it can realize large-scale industrial production of mechanization, automation, standardization and cleanliness. The invention creates non-bamboo series plant functional salt represented by pine salt and plum salt, which is a revolution to the table salt industry (especially the high-end salt market) and is of great significance to the health cause of all human beings .
为解决上述技术问题，本发明提出一种功能性植物盐的制备方法，包括以下步骤：To solve the above technical problems, the present invention proposes a method for preparing a functional plant salt, which includes the following steps:
将一种或多种植物提取物粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温下一次或分次烤制，获得功能性植物盐。One or more plant extract powders are mixed with fine salt uniformly at an added amount of 1% to 40%, and roasted at a high temperature of 800-1400 ° C once or in stages to obtain a functional plant salt.
作为本发明功能性植物盐的制备方法的改进：As an improvement of the preparation method of the functional plant salt of the present invention:
所述植物提取物粉末为一种时：When the plant extract powder is one type:
将一种植物提取物粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温下一次烤制，获得功能性植物盐。A plant extract powder is uniformly mixed with fine salt at an addition amount of 1% to 40%, and is roasted at a high temperature of 800-1400 ° C at one time to obtain a functional plant salt.
作为本发明功能性植物盐的制备方法的进一步改进：As a further improvement of the preparation method of the functional plant salt of the present invention:
所述植物提取物粉末为多种(至少两种)时：When the plant extract powder is multiple (at least two):
将多种植物提取物粉末混合均匀后，获得混合粉末；After mixing a plurality of plant extract powders uniformly, a mixed powder is obtained;
将所得混合粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温下一次烤制，获得功能性植物盐。The obtained mixed powder is uniformly mixed with fine salt at an addition amount of 1% to 40%, and is roasted at a high temperature of 800-1400 ° C at one time to obtain a functional plant salt.
作为本发明功能性植物盐的制备方法的进一步改进：As a further improvement of the preparation method of the functional plant salt of the present invention:
所述植物提取物粉末为多种(至少两种)时：When the plant extract powder is multiple (at least two):
S1、将一种植物提取物粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温烤制并保温一段时间；S1. Mix a kind of plant extract powder with fine salt according to the addition amount of 1% to 40%, bake it at a high temperature of 800-1400 ° C and keep it for a while;
S2、将步骤S1中于800-1400℃保温一定时间的所得物取出，粉碎后再次向其中添加另一种植物提取物粉末，重复步骤S1和S2(注：此时在原温度或更高的温度下继续烤制)，直至所有植物提取物粉末烤制完成，获得功能性植物盐。S2. Take out the product that was kept at 800-1400 ° C for a certain time in step S1, add another plant extract powder to it after crushing, and repeat steps S1 and S2 (Note: at this time, the original temperature or higher Continue to roast) until all the plant extract powder is roasted to obtain a functional plant salt.
为解决上述技术问题，本发明还提出一种利用上述制备方法制备所得的功能性植物盐。In order to solve the above technical problems, the present invention also provides a functional plant salt prepared by using the above preparation method.
为解决上述技术问题，本发明还提出一种功能性植物盐的用途：In order to solve the above technical problems, the present invention also proposes the use of a functional plant salt:
所述功能性植物盐用于药物助剂、保健食品/饮品、功能性食品/饮品、餐桌 调味盐、烹饪盐、特殊用途化妆品或个人护理用品。The functional plant salt is used in pharmaceutical auxiliaries, health foods / drinks, functional foods / beverages, table seasoning salts, cooking salts, special-purpose cosmetics or personal care products.
本发明所述的功能性植物盐，其特征是，采用中频熔炼技术或电阻加热(又称欧姆加热)技术，将不同来源的竹提取物与食盐按一定比例混匀后，于常压下在800～1400℃之间熔炼，冷却后重结晶，得到的新型竹盐，可广泛应用于食品、饮品、调味品、保健食品、药品、日用化学品、个人护理用品等。本发明首创的以松盐和梅盐为代表的非竹植物功能盐除了有着与竹盐相同的生物学效应外，还具有预防高血压、美白、护肝等多种作用。The functional plant salt according to the present invention is characterized in that, by using intermediate frequency melting technology or resistance heating (also known as ohmic heating) technology, bamboo extracts from different sources are mixed with table salt in a certain ratio, The new bamboo salt obtained by smelting at 800 ～ 1400 ℃ and recrystallizing after cooling can be widely used in food, beverages, condiments, health foods, pharmaceuticals, daily chemicals, personal care products, etc. The non-bamboo plant functional salt represented by pine salt and plum salt, which is pioneered by the present invention, has the same biological effects as bamboo salt, and also has various functions such as preventing hypertension, whitening, and protecting the liver.
根据竹提取物的种类、来源及其有效成分、添加比例、烤制工艺参数等的不同，得到不同色泽、不同晶格结构、不同理化指标、不同风味和不同用途的新型竹盐，其产品突出的特征和共性是：盐溶液由弱酸性变为强碱性，色泽由白色变为灰色、浅灰色、淡粉色、紫红色和亮蓝色等不同颜色，呈现出晶莹剔透的干净外观，并伴有淡淡的硫味；盐的晶胞尺寸较原盐更小，盐的元素组成更为丰富，其中钾含量提高约100倍，钙约7倍，其他有益元素也显著增加。According to the different types, sources and effective components of bamboo extracts, addition ratios, and roasting process parameters, new bamboo salts with different colors, different lattice structures, different physical and chemical indexes, different flavors, and different uses are obtained. The products are outstanding. The characteristics and common features are: the salt solution changes from weakly acidic to strongly alkaline, and the color changes from white to gray, light gray, light pink, fuchsia, and bright blue, showing a clean and clear appearance, accompanied by There is a slight sulphur smell; the unit cell size of the salt is smaller than that of the original salt, and the elemental composition of the salt is more abundant, in which the potassium content is increased by about 100 times, calcium is about 7 times, and other beneficial elements are also significantly increased.
所述的中频熔炼技术，是将工频50Hz的交流电转变为中频(300～1000Hz)，把三相工频交流电整流后变成直流电，再把直流电变为可调节的中频电流，供给由电容和感应线圈里流过的中频交变电流，在感应圈中产生高密度的磁力线，通过导磁的容器(如石墨碳化硅坩埚)迅速生产高温，以此加热物料(即盐与提取物的混合物)使其熔融。可以在短时间内迅速达到熔炼温度，大幅度提高生产效率。The intermediate frequency melting technology is to convert AC power of 50Hz to intermediate frequency (300 ~ 1000Hz), rectify the three-phase power frequency AC to DC, and then convert DC to adjustable intermediate frequency current. The intermediate frequency alternating current flowing in the induction coil generates high-density magnetic lines of force in the induction coil, and rapidly produces high temperature through a magnetically permeable container (such as a graphite silicon carbide crucible) to heat the material (that is, a mixture of salt and extract) Let it melt. The melting temperature can be reached quickly in a short time, which greatly improves the production efficiency.
具体工艺及参数如下：The specific process and parameters are as follows:
按0～40％的质量比(优选5～25％)，将不同的植物提取物粉末与食盐均匀混合；Mix different plant extract powders with table salt at a mass ratio of 0 to 40% (preferably 5 to 25%);
将以上混合物料放入导磁坩埚内、置于中频熔炼炉中，开始升温，升温速率为20～100℃/min；Put the above mixture in a magnetically-induced crucible and place it in an intermediate-frequency melting furnace, and begin to raise the temperature, with a heating rate of 20 to 100 ° C / min;
在常压下升至800～1400℃之间的某一温度区间、保温0～6h；Rise to a temperature range between 800 ～ 1400 ℃ under normal pressure, and keep it for 0 ～ 6h;
将溶融的物料倒出，自然降温或控制性降温；Pour out the melted material and cool it naturally or controlledly;
冷却结晶后，粉碎、过筛得到产品；After cooling and crystallization, crushing and sieving to obtain the product;
熔炼温度可设置在800～1400℃之间的1个或多个区间内；The melting temperature can be set in one or more intervals between 800 and 1400 ° C;
以上混合—熔融—冷却—重结晶—粉碎的过程可一次或多次进行。The above mixing-melting-cooling-recrystallization-pulverizing process can be performed one or more times.
电阻加热(又称欧姆加热)技术，是将电能转化为热能，从而对物料进行快速加热。电阻炉和火焰炉比，热效率高，温度容易控制，分为直接加热和间接加热两种方式。目前大部分电阻炉采用间接加热，设备中装有实现电-热转变的电阻元件，称为电热体，由它把热能传给待加工的物料。常用的电热体有碳化硅棒和二硅化钼棒，其中硅碳棒的温度可达1400℃，硅钼棒可达1700℃。Resistance heating (also known as ohmic heating) technology converts electrical energy into thermal energy to rapidly heat materials. The ratio of resistance furnace to flame furnace has high thermal efficiency and easy temperature control. It is divided into direct heating and indirect heating. At present, most resistance furnaces use indirect heating, and the equipment is equipped with a resistance element that realizes electric-thermal conversion, called an electric heating body, which transfers heat energy to the material to be processed. Commonly used electric heating bodies are silicon carbide rods and molybdenum disilicide rods. The temperature of silicon carbon rods can reach 1400 ° C, and silicon molybdenum rods can reach 1700 ° C.
具体工艺及参数如下：The specific process and parameters are as follows:
按0～40％的质量比(优选5～25％)，将不同的植物提取物粉末与食盐均 匀混合；Mix different plant extract powders with table salt at a mass ratio of 0 to 40% (preferably 5 to 25%);
将以上混合物料放入刚玉坩埚内、置于电阻炉中，开始升温，升温速率为1～20℃/min；Put the above mixture into the corundum crucible and put it in the resistance furnace, and start to raise the temperature, the heating rate is 1 ～ 20 ℃ / min;
在常压下升至800～1400℃之间的某一温度保温0～6h；Rise to a temperature between 800 ～ 1400 ℃ under normal pressure for 0 ～ 6h;
将物料倒出，自然降温或控制性降温；Pour out the material and cool it naturally or in a controlled way;
冷却结晶后，粉碎、过筛得到产品；After cooling and crystallization, crushing and sieving to obtain the product;
熔炼温度可设置在800～1400℃之间的1个或多个区间内；The melting temperature can be set in one or more intervals between 800 and 1400 ° C;
以上混合—熔融—重结晶—冷却—粉碎的过程可一次或多次进行。The above mixing-melting-recrystallization-cooling-pulverizing process can be performed one or more times.
所述的植物提取物，又称植物化学素，即为植物的次生代谢产物。包含各种水果、蔬菜、花卉、中草药、药食两用植物以及源于植物的新资源食品的植物粗提物、高精度制剂乃至单体化合物，如竹、梅、松、茶、荷、菊、石榴、海棠、食用菌、藻类提取物等。同一植物来源还可以是其不同部位的提取物，如竹取物包括竹叶提取物(如竹叶黄酮、竹叶多酚)、竹茹提取物(如竹茹三萜、竹茹多糖)、竹竿提取物、竹笋提取物(如竹笋甾醇、竹笋氨基酸肽)，梅取物包括梅果提取物、梅花提取物、梅叶提取物、梅枝提取物、梅根提取物、梅核提取物，松取物包括松针提取物、松树皮提取物、松花粉提取物。The plant extract, also known as a phytochemical, is a secondary metabolite of a plant. Contains a variety of fruits, vegetables, flowers, Chinese herbs, medicinal and edible plants, and plant-derived new resource foods, crude plant extracts, high-precision preparations and even monomer compounds such as bamboo, plum, pine, tea, lotus, chrysanthemum , Pomegranate, begonia, edible fungus, algae extract, etc. The same plant source can also be extracts from different parts of it, such as bamboo extracts including bamboo leaf extracts (such as bamboo leaf flavones, bamboo leaf polyphenols), bamboo rut extracts (such as ruta triterpenes, ruta polysaccharides), Bamboo pole extract, bamboo shoot extract (such as bamboo shoot sterol, bamboo shoot amino acid peptide), plum extracts include plum fruit extract, plum extract, plum leaf extract, plum branch extract, plum root extract, plum kernel extract, pine The extracts include pine needle extract, pine bark extract, and pine pollen extract.
根据植物提取物种类和来源的不同，将植物功能盐分成竹和非竹二大系列。应用竹取物系列制得的归为竹功能盐，竹以外来源的则统称非竹功能盐，包括用藻类、蕈类(食用菌)及微生物来源的提取物(或称抽提物)制备得到的功能盐。According to the different types and sources of plant extracts, plant functional salts are divided into two series: bamboo and non-bamboo. It is classified as bamboo functional salt by using bamboo extract series. Non-bamboo functional salts are collectively referred to as non-bamboo functional salts. They include algae, mushrooms (edible fungi) and microbial-derived extracts (or extracts). Functional salt.
所述的食盐原料(即原盐)，可以是海盐、湖盐、井盐、矿盐，优选粗制的海盐(又称日晒海盐)。The common salt raw material (that is, raw salt) may be sea salt, lake salt, well salt, mineral salt, preferably crude sea salt (also known as sun-sea salt).
所述的非竹功能盐的健康功效包括但不限于：预防高血压、美白、护肝。The health effects of the non-bamboo functional salts include, but are not limited to, preventing hypertension, whitening, and protecting the liver.
所述的功能性植物盐的应用领域包括：食品、饮品、调味品、保健食品、药品、日用化学品和个人护理用品。The application fields of the functional plant salt include: food, beverages, condiments, health foods, medicines, daily chemicals and personal care products.
采用本发明的中频熔炼技术或电阻加热技术也可直接加工食盐(即不添加植物提取物或任何其他外来物质)，将其转化为食用安全性更高的烤盐。The intermediate frequency melting technology or the resistance heating technology of the present invention can also directly process table salt (that is, without adding plant extracts or any other foreign substances), and convert it into roasted salt with higher food safety.
本发明与现有技术相比，具有如下技术优势：Compared with the prior art, the present invention has the following technical advantages:
本发明制备所得功能性植物盐的具体的生物学功能包括：抑菌、抗炎、抗病毒；护肝益肾、安神健胃、排毒解酒、止血消痔；治疗偏头痛、过敏性鼻炎、对人牙龈成纤维细胞的抗炎作用、减肥功能、保护口腔健康、抑制结肠癌变、增加抗突变和体外抗癌作用、HepG2人肝癌细胞的抗突变活性和体外抗癌作用、预防神经细胞凋亡等。此外，有助于提升人体表皮细胞的渗透性，帮助日用化学品中的护肤因子更好地起作用。The specific biological functions of the functional plant salt prepared by the present invention include: bacteriostatic, anti-inflammatory, anti-viral; protecting the liver and kidney, soothing the stomach, detoxifying and detoxifying, stopping bleeding and hemorrhoids; treating migraine, allergic rhinitis, Anti-inflammatory effect on human gingival fibroblasts, weight loss function, protecting oral health, inhibiting colon cancer, increasing anti-mutation and anti-cancer effects in vitro, anti-mutation activity of HepG2 human liver cancer cells and anti-cancer effects in vitro, preventing neuronal apoptosis Wait. In addition, it helps to improve the permeability of human epidermal cells and helps skin care factors in daily chemicals work better.
本发明制备所得功能性植物盐的具体用途包括：药物助剂；保健食品/饮品；功能性食品/饮品；餐桌调味盐，烹饪盐；特殊用途化妆品，个人护理用品(护肤 霜、粉底液、香皂、面膜、牙膏、漱口水、洗发液、沐浴盐等)；终端产品的形态可以是粉末、胶囊、片剂、颗粒剂、泡腾片、水剂、乳剂、喷雾剂等。Specific uses of the functional plant salt prepared by the present invention include: pharmaceutical auxiliaries; health food / drinks; functional foods / drinks; table seasoning salt, cooking salt; special-purpose cosmetics, personal care products (skin cream, liquid foundation, soap , Facial mask, toothpaste, mouthwash, shampoo, bath salt, etc.); the form of the end product can be powder, capsule, tablet, granule, effervescent tablet, water, emulsion, spray, etc.
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。The present invention will be further described below with reference to specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions in the following examples are usually performed according to the conventional conditions or the conditions recommended by the manufacturer. Unless stated otherwise, percentages and parts are by weight.
除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equal to those described can be used in the method of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.
通用原料和设备General materials and equipment
竹叶黄酮：棕色粉末，购自浙江圣氏生物科技有限公司(安吉)，总黄酮含量为24.8％；Bamboo leaf flavonoids: brown powder, purchased from Zhejiang Shengshi Biotechnology Co., Ltd. (Anji), with a total flavonoid content of 24.8%;
竹笋甾醇：黄色粉末，由浙江大学生物系统工程与食品科学学院的“天然产物与人类健康”研究中心提供，总甾醇含量为25％；Bamboo shoot sterols: yellow powder, provided by the "Natural Products and Human Health" Research Center of the College of Biological System Engineering and Food Science, Zhejiang University, with a total sterol content of 25%;
竹茹三萜：黄绿色粉末，由浙江大学生物系统工程与食品科学学院的“天然产物与人类健康”研究中心提供，总三萜皂苷含量为45％；Zhuru triterpenes: yellow-green powder, provided by the "Natural Products and Human Health" research center of the College of Biological System Engineering and Food Science, Zhejiang University, with a total triterpenoid saponin content of 45%;
竹笋氨基酸肽：灰色粉末，由浙江大学生物系统工程与食品科学学院的“天然产物与人类健康”研究中心提供，总氨基酸含量为21％；Bamboo shoot amino acid peptide: gray powder, provided by the "Natural Product and Human Health" Research Center of the College of Biological System Engineering and Food Science, Zhejiang University, with a total amino acid content of 21%;
竹多糖：由湖南城步锦龙农林科技开发有限责任公司提供，灰白色粉末，多糖含量为40％。Bamboo polysaccharide: provided by Hunan Chengbu Jinlong Agriculture and Forestry Technology Development Co., Ltd., an off-white powder with a polysaccharide content of 40%.
松针提取物：棕色粉末，购自陕西清雅生物科技有限公司；Pine needle extract: brown powder, purchased from Shaanxi Qingya Biotechnology Co., Ltd .;
梅枝提取物：黄色粉末，由杭州尤美特生物科技有限公司提供；Plum branch extract: yellow powder, provided by Hangzhou Youmeite Biotechnology Co., Ltd .;
原盐(日晒海盐)：来自山东潍坊，实测NaCl含量为93.36％；Raw salt (sun-sea salt): from Weifang, Shandong, the measured NaCl content is 93.36%;
中频熔炼炉：加热元件为紫铜感应线圈，坩埚为导磁的石墨碳化硅坩埚；Medium frequency melting furnace: the heating element is a copper induction coil, and the crucible is a magnetically conductive graphite silicon carbide crucible;
管式电阻炉：加热元件为硅碳棒。Tube resistance furnace: The heating element is a silicon carbon rod.
实施例1、功能性植物盐的制备方法，包括以下步骤：Example 1. A method for preparing a functional plant salt, including the following steps:
1、获取植物提取物粉末：1. Obtain plant extract powder:
上述植物提取物包括但不限于各种水果、蔬菜、食用花卉、中草药和药食两用植物的植物提取物，以及源于植物的新资源食品等，如竹、梅、松、茶、荷、 菊、石榴、海棠、食用菌、藻类提取物等。The above-mentioned plant extracts include, but are not limited to, plant extracts of various fruits, vegetables, edible flowers, Chinese herbal medicine and medicinal food plants, as well as new sources of food derived from plants, such as bamboo, plum, pine, tea, lotus, Daisy, pomegranate, begonia, edible fungus, algae extract, etc.
同一植物来源还可以是其不同部位的提取物、如竹取物包括竹叶提取物(竹叶黄酮、竹叶多酚)、竹茹提取物、竹竿提取物、竹笋提取物，梅取物包括梅花提取物、梅果提取物、梅叶提取物、梅枝提取物、梅根提取物，松取物包括松针提取物、松树皮提取物、松花粉提取物等。The same plant source can also be extracts from different parts of it, such as bamboo extracts including bamboo leaf extracts (bamboo leaf flavones, bamboo leaf polyphenols), bamboo rut extracts, bamboo pole extracts, bamboo shoot extracts, and plum extracts including Plum blossom extract, plum fruit extract, plum leaf extract, plum branch extract, plum root extract, pine extracts include pine needle extract, pine bark extract, pine pollen extract, and the like.
2、将步骤1中植物提取物粉末按1％～40％的添加量与细盐混合均匀，获得混合物；2. Mix the plant extract powder in step 1 with the fine salt in an amount of 1% to 40% to obtain a mixture;
将所得混合物放入特制的容器内，于中频熔炼炉(或其他机械化烤炉)中，在一定的升温速率下加热到800-1400℃，获得功能性植物盐。The obtained mixture is put into a special container, and is heated to 800-1400 ° C. in a medium-frequency melting furnace (or other mechanized oven) at a certain heating rate to obtain a functional plant salt.
上述细盐的来源可以是海盐、湖盐、井盐、矿盐，优选粗制海盐。The source of the fine salt may be sea salt, lake salt, well salt, mineral salt, and preferably crude sea salt.
上述％为质量比。The above% is a mass ratio.
实施例2、将实施例1中植物提取物粉末更改为至少两种植物提取物粉末，即，将至少两种植物提取物粉末按任意比例混合均匀后，获得混合粉末；将混合粉末按1％～40％的添加量与细盐(过筛)混合均匀，获得混合物；其余等同于实施例1。Example 2: The plant extract powder in Example 1 was changed to at least two plant extract powders, that is, the at least two plant extract powders were mixed uniformly at any ratio to obtain a mixed powder; the mixed powder was 1% The added amount of -40% is mixed with fine salt (sieved) uniformly to obtain a mixture; the rest is equivalent to Example 1.
实施例3、将实施例1中植物提取物粉末更改为至少两种植物提取物粉末(其中一种为竹取物)，具体步骤如下： Embodiment 3. The plant extract powder in Example 1 is changed to at least two plant extract powders (one of which is bamboo extract), and the specific steps are as follows:
1、将一种植物提取物粉末与细盐混合均匀，获得混合物；1. Mix a plant extract powder with fine salt to obtain a mixture;
2、将所得混合物放入特制的容器内，于中频熔炼炉(或其他机械化烤炉)中，在一定的升温速率下加热到800-1400℃，并保温一定时间，获得所得物；2. Put the obtained mixture into a special container, heat it in an intermediate frequency melting furnace (or other mechanized oven) to 800-1400 ℃ at a certain heating rate, and keep it for a certain time to obtain the product;
3、将步骤2中于800-1400℃保温一定时间的所得物取出，粉碎后再次向其中添加另一种植物提取物粉末，重复步骤1-3(注：此时在原温度或更高的温度下继续烤制)，直至所有植物提取物粉末烤制完成，获得功能性植物盐3. Take out the product that was kept at 800-1400 ℃ for a certain period of time in step 2 and add another plant extract powder to it after crushing, and repeat steps 1-3 (Note: at this time at the original temperature or higher Continue to roast) until all plant extract powder is roasted to obtain functional plant salt
注：上述所采用植物提取物粉末的总添加量为1％～40％(质量比)。Note: The total added amount of the plant extract powder used above is 1% to 40% (mass ratio).
综上，本发明的功能性植物盐既可以是添加植物提取物后一次性烤制完成，也可以是分次添加、分阶段完成烤制。In summary, the functional plant salt of the present invention can be roasted at one time after adding plant extracts, or it can be roasted in stages and added in stages.
本发明提供的功能性植物盐，在高温熔炼过程中，由于不同植物提取物中不同矿物元素的渗入，元素组成和NaCl晶格、晶体构型都发生了显著的变化。根据不同的添加比例和烤制方式，呈现出不同的颜色，如灰色、紫色、粉色、白色、蓝色、青绿色等等；5％盐水溶液的pH值从原料盐的6-7升高至8-12；NaCl总量下降，其他元素(尤其是多种微量元素)含量增加；口感从单纯的咸味转变为丰富圆润的咸鲜甜味，还带有显著的臭皮蛋味。During the high-temperature smelting process of the functional plant salt provided by the present invention, due to the infiltration of different mineral elements in different plant extracts, the elemental composition, NaCl crystal lattice, and crystal configuration have undergone significant changes. According to different addition ratios and roasting methods, different colors are displayed, such as gray, purple, pink, white, blue, turquoise, etc .; the pH value of the 5% saline solution is increased from 6-7 of the raw salt to 8-12; The total amount of NaCl decreases, and the content of other elements (especially a variety of trace elements) increases; the taste changes from a simple salty taste to a rich and rounded savory sweetness, and it also has a significant rotten egg smell.
例如，添加松、竹、梅提取物后获得的“松盐”、“竹盐”和“梅盐”，由于提取物的来源、化学组成及添加量的不同，以及烤制次数和工艺参数的不同，所获得的植物盐的理化性质、外观色泽、风味口感存在显著差异，由此带来的生物学效应也有不同。For example, the "pine salt", "bamboo salt", and "plum salt" obtained after adding pine, bamboo, and plum extracts are different due to the origin of the extract, the chemical composition and the amount of addition, and the number of roasting times and process parameters The physical and chemical properties, appearance, color, and flavor of the obtained plant salt are significantly different, and the biological effects caused by them are also different.
功能性植物盐的具体的生物学功能包括：抑菌、抗炎、抗病毒；护肝益肾、安神健胃、排毒解酒、止血消痔；治疗偏头痛、过敏性鼻炎、对人牙龈成纤维细胞的抗炎作用、减肥功能、保护口腔健康、抑制结肠癌变、增加抗突变和体外抗癌作用、HepG2人肝癌细胞的抗突变活性和体外抗癌作用、预防神经细胞凋亡等。此外，有助于提升人体表皮细胞的渗透性，帮助日用化学品中的护肤因子更好地起作用。The specific biological functions of functional plant salts include: bacteriostatic, anti-inflammatory, anti-viral; liver-protecting kidney, soothe the stomach, detoxify and detoxify alcohol, stop bleeding and relieve hemorrhoids; treat migraine, allergic rhinitis, and affect human gums Fibrocytes have anti-inflammatory effects, weight loss functions, protect oral health, inhibit colon canceration, increase anti-mutation and anti-cancer effects in vitro, anti-mutation activity of HepG2 human liver cancer cells and anti-cancer effects in vitro, and prevent neuronal apoptosis. In addition, it helps to improve the permeability of human epidermal cells and helps skin care factors in daily chemicals work better.
功能性植物盐的具体用途包括：作为药物助剂；保健食品/饮品；功能性食品/饮品；餐桌调味盐，烹饪盐；特殊用途化妆品，和人护理用品(护肤霜、粉底液、香皂、面膜、牙膏、漱口水、洗发液、沐浴盐等)；终端产品的形态可以是粉末、胶囊、片剂、颗粒剂、泡腾片、水剂、乳剂、喷雾剂等。Specific uses of functional plant salts include: as pharmaceutical auxiliaries; health foods / drinks; functional foods / drinks; table seasoning salts, cooking salts; special-purpose cosmetics, and human care products (skin creams, liquid foundations, soaps, masks , Toothpaste, mouthwash, shampoo, bath salt, etc.); the form of the end product can be powder, capsule, tablet, granule, effervescent tablet, water, emulsion, spray and so on.
综上，本发明提出了一种功能性植物盐的制备方法，将不同的植物提取物与食盐按一定比例混合均匀后，在800-1400℃的高温下一次或分次烤制所形成的具有不同颜色、不同空间结构、不同理化性质、不同风味和不同用途的功能性植物盐。本发明制备所得的功能性植物盐不仅具有保健作用，还有助于提升人体表皮细胞的渗透性，帮助日用化学品中的护肤因子更好地起作用，可用于药物助剂、保健食品/饮品、功能性食品/饮品、餐桌调味盐、烹饪盐、特殊用途化妆品或护理用品。In summary, the present invention provides a method for preparing a functional plant salt. Different plant extracts and table salt are mixed uniformly at a certain ratio, and they are formed by roasting one or several times at a high temperature of 800-1400 ° C. Functional plant salts of different colors, different spatial structures, different physical and chemical properties, different flavors and different uses. The functional plant salt prepared by the invention not only has a health-care effect, but also helps to improve the permeability of human epidermal cells, and helps the skin care factors in daily chemicals to function better. It can be used in pharmaceutical auxiliaries, health foods / Beverages, functional foods / drinks, table salt, cooking salt, special purpose cosmetics or care products.
实施例4、植物性功能盐—松盐的制作(中频熔炼)Example 4 Production of Plant Functional Salts—Pine Salt (Intermediate Frequency Smelting)
分别称取6.0g松针提取物和24.0g原盐，混合均匀后放至导磁坩埚内，置于中频熔炼炉中，在常压下按100℃/min的升温速率升至1000℃，在1000℃下保温1h后取出自然降温，得到粉紫色的松盐(代码为Plum Salt)，其色泽和外观如图1所示。Weigh 6.0g of pine needle extract and 24.0g of raw salt, mix them and put them into a magnetic crucible, put them in an intermediate frequency melting furnace, and raise them to 1000 ℃ at a normal temperature of 100 ℃ / min, at 1000 ℃ After incubation for 1 hour, the temperature was taken out to cool down to obtain pink-purple pine salt (code: Plum Salt). Its color and appearance are shown in Figure 1.
实施例5、植物性功能盐—新型紫竹盐的制作(中频熔炼)Example 5. Production of botanical functional salt—new purple bamboo salt (medium frequency melting)
分别称取竹叶提取物10g和原盐40g，混合均匀后放入导磁坩埚内，置于中频熔炼炉中，在常压下按100℃/min的升温速率升至900℃后保温1.5h，取出自然降温，冷却后粉碎得到新型的紫竹盐，其色泽和外观如图2所示。Weigh 10g of bamboo leaf extract and 40g of raw salt respectively, mix them well and put them in a magnetic crucible, put them in an intermediate frequency melting furnace, and raise the temperature to 900 ° C at a temperature of 100 ° C / min under normal pressure for 1.5h. Take out the natural cooling, pulverize after cooling to obtain a new type of purple bamboo salt, its color and appearance are shown in Figure 2.
实施例6、植物性功能盐—梅盐的制作(中频熔炼)Example 6 Preparation of Plant Functional Salt-Plum Salt (Medium Frequency Smelting)
分别称取5.0g梅枝提取物和25.0g原盐，混合均匀后放入导磁坩埚，置于中频熔炼炉中，在常压下按100℃/min的升温速率升至1300℃，在1300℃下 保温1h后取出自然降温，得到蓝色的梅盐(记为Plum Salt)，其色泽和外观如图3所示。Weigh 5.0g of plum branch extract and 25.0g of raw salt, mix well and put them into a magnetic crucible, put them in an intermediate frequency melting furnace, and raise it to 1300 ℃ at a normal temperature of 100 ℃ / min, at 1300 ℃ After holding for 1 hour, the temperature was taken out to cool down to obtain blue plum salt (recorded as Plum Salt). Its color and appearance are shown in Figure 3.
实施例7、植物性功能盐—松盐的制作(电阻加热)Example 7 Preparation of Plant Functional Salt—Pine Salt (Resistance Heating)
分别称取4.0g松针提取物和26.0g原盐，混合均匀后放至导磁坩埚内，置于管式电阻炉中，在常压下按100℃/min的升温速率升至1100℃，在1100℃下保温2h后取出自然降温，得到粉紫色的松盐，其色泽和外观如图4所示。Weigh 4.0g of pine needle extract and 26.0g of raw salt, mix them and place them in a magnetic crucible, put them in a tube resistance furnace, and raise them to 1100 ℃ at a temperature of 100 ℃ / min under normal pressure. After incubating at ℃ for 2 hours, the temperature was taken out and the temperature was reduced to obtain a pink purple pine salt. The color and appearance are shown in FIG. 4.
实施例8、植物性功能盐—新型紫竹盐的制作(电阻加热)Example 8. Production of botanical functional salt-new type of purple bamboo salt (resistance heating)
分别称取竹叶提取物15g和原盐35g，混合均匀后放入刚玉坩埚内，置于管式炉中，在常压下按10℃/min的升温速率升至950℃后保温2.5h，取出自然降温，冷却后粉碎得到紫竹盐，其色泽和外观如图5所示。Weigh 15g of bamboo leaf extract and 35g of raw salt, mix them well, put them into a corundum crucible, put them in a tube furnace, raise the temperature to 950 ℃ at a temperature of 10 ℃ / min under normal pressure, and keep them for 2.5h. The natural temperature is lowered, and the shredded bamboo salt is obtained after cooling, and its color and appearance are shown in FIG. 5.
实施例9、植物性功能盐—梅盐的制作(电阻加热)Example 9 Production of Plant Functional Salt-Plum Salt (Resistance Heating)
分别称取8.0g梅枝提取物和22.0g原盐，混合均匀后放入刚玉坩埚，置于管式电阻炉中，在常压下按10℃/min的升温速率升至1250℃，在1250℃下保温1.5h后取出自然降温，得到蓝色的梅盐，其色泽和外观如图6所示。Weigh 8.0g of plum branch extract and 22.0g of raw salt respectively, mix them well, put them into a corundum crucible, put them in a tube resistance furnace, and raise it to 1250 ° C at a heating rate of 10 ° C / min under normal pressure, at 1250 ° C After holding for 1.5 hours, the temperature was taken out to cool down to obtain blue plum salt. The color and appearance are shown in Figure 6.
性能测试(实验中松、竹、梅盐分别为实施例4、5、6所得样品)Performance test (In the experiment, pine, bamboo, and plum salt were the samples obtained in Examples 4, 5, and 6, respectively)
实验1、植物功能盐及其对照品的理化性质分析 Experiment 1. Analysis of Physicochemical Properties of Plant Functional Salts and Their References
1.1色差分析1.1 Color difference analysis
植物提取物种类、添加比例、熔炼温度、保温时间乃至升温速率都会对终产品的色泽和外观产生直接影响。图7显示的添加5～35％不同植物提取物的原盐，在常压下经过800～1400℃下1～2h的高温熔炼，实施例4所得松盐、实施例5所得竹盐和实施例6所得梅盐的外观，分别可呈现灰色、浅灰色、淡粉色、蓝紫色和亮蓝色等不同色泽。同时，图7中可见，不添加任何外来物质的原盐经过800～900℃下1～2h的高温熔炼后得到的烤盐透明度更高，色泽更亮。The type of plant extract, the addition ratio, the melting temperature, the holding time and even the heating rate will directly affect the color and appearance of the final product. Figure 7 shows raw salt added with 5 to 35% of different plant extracts, subjected to high temperature smelting at 800 to 1400 ° C for 1 to 2 hours under normal pressure, the pine salt obtained in Example 4, the bamboo salt obtained in Example 5, and Example 6 The appearance of the obtained plum salt can show different colors such as gray, light gray, pale pink, blue-violet and bright blue. At the same time, it can be seen in FIG. 7 that the roasted salt obtained by smelting the raw salt without adding any foreign substances at a high temperature of 800 to 900 ° C. for 1 to 2 hours has higher transparency and brighter color.
色差仪是一种常见的光电积分式测色仪器，它仿照人眼感色的原理，采用能感受红、绿、蓝三种颜色的受光器，利用仪器内部的标准光源照射被测物体，将各自所感受的光电流加以放大处理，在整个可见光波长范围内进行一次积分测量，得到透射或反射物体色的三刺激值和色品坐标，从而获得这一颜色的信号，并通过系统给出被测样品之间的色差值。The color difference meter is a common photoelectric integrating color measuring instrument. It imitates the principle of color perception of the human eye. It uses a light receiver that can sense three colors of red, green, and blue. The standard light source inside the instrument is used to illuminate the measured object. The photocurrents experienced by each are amplified, and an integral measurement is performed over the entire visible light wavelength range to obtain the tristimulus value and chromaticity coordinates of the color of the transmitted or reflected object, thereby obtaining the signal of this color, and giving the signal by the system. Measure the color difference between samples.
色差仪测得的主要指标包括L*、a*、b*。其中，The main indicators measured by the color difference meter include L *, a *, b *. among them,
L*表示试样的亮度，数值越大，表明越亮；L * represents the brightness of the sample, the larger the value, the brighter it is;
a*代表红绿色向，“+”表示偏向红色，“-”表示偏向绿色；a * stands for red and green, "+" means red, and "-" means green.
b*代表蓝黄色相，“+”表示偏向黄色，“-”表示偏向蓝色。b * represents a blue-yellow phase, "+" indicates a bias toward yellow, and "-" indicates a bias toward blue.
ΔL值表示样品值与基准点的差值；Δa值表示样品的a*值相对基准点的差值，能更好的表示样品的红绿色值。The ΔL value represents the difference between the sample value and the reference point; the Δa value represents the difference between the a * value of the sample and the reference point, which can better represent the red-green value of the sample.
在本发明中，基准点指的是标样的值，本发明用标准白板作为标样。In the present invention, the reference point refers to the value of the standard sample, and the present invention uses a standard white board as the standard sample.
表1是不同植物功能盐的色差分析数据汇总。从中可见，原盐和烤盐ΔL值分别为84.18±0.25和82.83±3.47，显示其接近白色；而其他植物功能盐的ΔL值介于25～50之间，最暗的是仁山竹盐，最亮的是梅盐。不同试样的a*值和b*值差异显著(p<0.05)，表明其色向差异较大，呈现出截然不同的色相。Table 1 is a summary of the color difference analysis data of different plant functional salts. It can be seen that the ΔL values of raw salt and roasted salt are 84.18 ± 0.25 and 82.83 ± 3.47, showing that they are close to white; while the ΔL values of other plant functional salts are between 25-50, the darkest is the mangosteen salt, the brightest It's plum salt. The a * value and b * value of different samples are significantly different (p <0.05), which indicates that the color direction of them differs greatly, showing a completely different hue.
表1不同植物功能盐试样的色值参数Table 1 Color value parameters of different plant functional salt samples
Figure PCTCN2019097358-appb-000001
Figure PCTCN2019097358-appb-000001
1.2pH的变化1.2 pH change
食物的pH值对维持人体内环境的酸碱平衡有重要影响，故有酸性食物和碱性食物之的分。现代社会，大众的饮食结构普遍偏酸性，久而久之会使形成酸性体质，导致免疫力下降和亚健康状态。测得0.2～5％浓度(w/v)下不同盐样的pH值，如表2所示。The pH value of food has an important effect on maintaining the acid-base balance of the human body environment, so there is a distinction between acidic foods and alkaline foods. In modern society, the dietary structure of the general public is generally acidic. Over time, it will form an acidic constitution, which will lead to a decline in immunity and a sub-healthy state. The pH values of different salt samples were measured at a concentration of 0.2 to 5% (w / v), as shown in Table 2.
1％及以下浓度的原盐溶液呈弱酸性(pH<7)，烤制后显著上升；而所有植物功能盐溶液即使在0.2％的浓度下都呈现强碱性(pH>9)，与原盐相比差异十分显著(p<0.001)。表明，原盐在与植物提取物共融后，有大量碱金属元素进入了NaCl的晶格结构中，或附着在其表面。The original salt solution with a concentration of 1% or less is weakly acidic (pH <7), and it rises significantly after baking; all the plant functional salt solutions show strong alkalinity (pH> 9) even at a concentration of 0.2%, which is in phase with the original salt. The difference is very significant (p <0.001). It is shown that after the original salt is fused with the plant extract, a large number of alkali metal elements have entered the lattice structure of NaCl or adhered to the surface.
表2不同植物功能盐试样的pH值Table 2 pH values of different plant functional salt samples
Figure PCTCN2019097358-appb-000002
Figure PCTCN2019097358-appb-000002
1.3电导率的变化1.3 Changes in conductivity
测得0.2～5％浓度(w/v)下不同试样的电导率数据，如表3所示。由表可知， 在人体可耐受的盐溶液浓度(＜1.0％)下，本发明的植物功能盐与原盐的电导率无显著差异。The conductivity data of different samples at 0.2 to 5% concentration (w / v) were measured, as shown in Table 3. As can be seen from the table, at a salt solution concentration (<1.0%) that can be tolerated by the human body, there is no significant difference in electrical conductivity between the plant functional salt and the original salt.
表3不同植物功能盐试样的电导率值Table 3 Conductivity values of different plant functional salt samples
Figure PCTCN2019097358-appb-000003
Figure PCTCN2019097358-appb-000003
1.4原子能谱分析1.4 Atomic Energy Spectrum Analysis
原子能谱不仅能够扫描拍摄物料的表面形态，还能检测出材料表面各种元素及其相对含量。将本发明制得的植物功能盐及其对照样(颗粒)置于原子能谱仪(日本日立公司，SU-8010)下进行样品表层的微区点、线、面的元素定性、半定量及定量分析(检出限为0.5％)，所得结果见表4。Atomic energy spectroscopy can not only scan and photograph the surface morphology of materials, but also detect various elements on the surface of materials and their relative contents. The plant functional salt prepared by the present invention and its reference sample (particles) are placed under an atomic energy spectrometer (Hitachi, Japan, SU-8010) to perform element qualitative, semi-quantitative, and quantitative analysis on the micro-area points, lines, and surfaces of the sample Analysis (detection limit: 0.5%), the results are shown in Table 4.
表4原子能谱分析得到的不同试样的元素组成(％)Table 4 Elemental composition of different samples obtained by atomic energy spectrum analysis (%)
Figure PCTCN2019097358-appb-000004
Figure PCTCN2019097358-appb-000004
1.5元素组成分析1.5 Elemental composition analysis
元素检测委托西安国联质量检测技术股份有限公司进行，结果见表5。Elemental testing was entrusted to Xi'an Guolian Quality Inspection Technology Co., Ltd. The results are shown in Table 5.
盐的主要成分为氯化钠，此外还含有少量的其他矿物质元素。精制程度越高的盐，氯化钠含量越高，其他营养元素就越少，于人体健康是不利的。The main ingredient of salt is sodium chloride, and it also contains small amounts of other mineral elements. The more refined the salt, the higher the sodium chloride content, the less other nutrients, which is not good for human health.
钾元素可以调节细胞内渗透压和体液酸碱平衡，参与细胞内糖和蛋白质的代谢，有助于维持神经健康、心跳规律正常，预防中风，并协助肌肉正常收缩。在摄入高钠盐导致高血压的情况下，钾盐或富钾的食物就具有降压作用。Potassium can regulate intracellular osmotic pressure and body fluid acid-base balance, participate in the metabolism of sugar and protein in the cell, help maintain nerve health, normal heartbeat, prevent stroke, and assist normal muscle contraction. In the case of high blood pressure caused by ingestion of high sodium, potassium or potassium-rich foods have a hypotensive effect.
镁是人体细胞内的主要阳离子，它可以影响钾离子和钙离子的转运，调控信号的传递，参与能量代谢、蛋白质和核酸的合成并进行催化酶的激活和抑制等。Magnesium is the main cation in human cells. It can affect the transport of potassium and calcium ions, regulate signal transmission, participate in energy metabolism, protein and nucleic acid synthesis, and activate and inhibit catalytic enzymes.
磷是组成遗传物质核酸的基本成分之一，同时磷和钙都是骨骼牙齿的重要构成材料。磷可以保持体内ATP代谢的平衡，调节体内的酸碱平衡，参与体内能量的代谢。Phosphorus is one of the basic components of nucleic acids that make up genetic material. At the same time, phosphorus and calcium are important building materials for bones and teeth. Phosphorus can maintain the balance of ATP metabolism in the body, regulate the acid-base balance in the body, and participate in the metabolism of energy in the body.
锰是SOD的辅酶，其缺乏可引起神经衰弱，影响智力发育，同时还将导致胰 岛素合成和分泌的降低，影响糖代谢。Manganese is a coenzyme of SOD. Its deficiency can cause neurasthenia and affect intellectual development. At the same time, it will also lead to a decrease in insulin synthesis and secretion and affect glucose metabolism.
钼是人体必不可少的一种微量元素，是动物体内肝、肠中黄嘌呤氧化酶、醛类氧化酶的基本成分之一，也是亚硫酸肝素氧化酶的组成成分。研究表明，钼还有明显防龋作用，且对尿结石的形成有强烈抑制作用，人体缺钼易患肾结石。Molybdenum is an essential trace element for the human body. It is one of the basic components of xanthine oxidase and aldehyde oxidase in the liver and intestines of animals. It is also a component of heparin sulfase oxidase. Studies have shown that molybdenum also has a significant anti-caries effect, and has a strong inhibitory effect on the formation of urinary stones. Humans lacking molybdenum are prone to kidney stones.
钒是人体必需的一种微量元素，一般认为，它可能有助于防止胆固醇蓄积、降血糖、降血压、防止龋齿、帮助制造红血球等。Vanadium is an essential trace element in the human body. It is generally believed that it may help prevent cholesterol accumulation, lower blood sugar, lower blood pressure, prevent dental caries, and help make red blood cells.
硒是人体必需的重要微量元素，中国营养学会将硒列为人体必需的15种营养素之一，一般认为硒与免疫力、衰老、生殖功能等紧密相关，缺硒是克山病的重要原因。Selenium is an essential trace element necessary for the human body. The Chinese Nutrition Association lists selenium as one of the 15 essential nutrients for the human body. It is generally believed that selenium is closely related to immunity, aging, and reproductive functions. Selenium deficiency is an important cause of Keshan disease.
铬也是一种必要的微量营养元素，在所有胰岛素调节活动中起着重要作用，是重要的血糖调节剂。同时，铬的缺乏还与近视的形成有一定关系。Chromium is also an essential micronutrient, which plays an important role in all insulin regulating activities and is an important blood glucose regulator. At the same time, the lack of chromium is also related to the formation of myopia.
硅主要集中于骨骼、肺、淋巴结、胰腺、肾上腺、指甲和头发之中，在主动脉、气管、肌腱、骨骼和皮肤等结缔组织中含量最高，对人体的骨健康具有特殊意义。然而，由于工业的发展及生活方式的改变，现代人体内的硅元素往往极为不足。Silicon is mainly concentrated in bones, lungs, lymph nodes, pancreas, adrenal glands, nails, and hair. It is the highest in connective tissues such as aorta, trachea, tendons, bones, and skin, and has special significance for human bone health. However, due to industrial development and changes in lifestyle, the silicon element in modern human bodies is often extremely insufficient.
从不同盐样的元素检测结果可知，植物功能盐与原盐(及其烤盐)有着十分显著的差异。松、竹、梅盐的钾含量极显著增加，尤其是实施例2、3得到的新型竹盐中钾元素是原盐的100倍以上，且比传统工艺生产的仁山竹盐高出14.9％；竹盐(包含新旧2种试样)的镁、铁、镍、锌、钼、钒的含显著高于原盐和其他植物盐，用新技术获得的松盐和梅盐中还检出了硒的存在。在所有试样中，梅盐拥有远高于其他试样的碘(28mg/kg)、硼(26.6mg/kg)、磷(424mg/kg)、铬(4mg/kg)和仅次于竹盐的锰含量(18mg/kg)。植物盐的硅含量显著高于原盐及烤盐，且新工艺的竹盐高于传统生产的仁山竹盐。同时，松盐与梅盐中还检出了硒元素。From the results of elemental testing of different salt samples, it can be seen that plant functional salt and original salt (and its roasted salt) have very significant differences. The potassium content of pine, bamboo, and plum salts has increased significantly, especially the potassium content of the new bamboo salt obtained in Examples 2 and 3 is more than 100 times that of the original salt, and it is 14.9% higher than that of Renshan bamboo salt produced by traditional processes; The salt (including the old and new samples) contained magnesium, iron, nickel, zinc, molybdenum, and vanadium significantly higher than the original salt and other plant salts. The selenium was also detected in the pine and plum salts obtained by the new technology. . Among all the samples, plum salt had much higher iodine (28mg / kg), boron (26.6mg / kg), phosphorus (424mg / kg), chromium (4mg / kg) and second only to bamboo salt than other samples. Manganese content (18mg / kg). The silicon content of plant salt is significantly higher than that of raw salt and roasted salt, and the bamboo salt of the new process is higher than the traditionally produced Renshan bamboo salt. At the same time, selenium was detected in pine and plum salts.
竹盐中还存在一定量的硫化物，带有典型的臭皮蛋气味。硫化氢有多种神经保护作用，已被国内外大量实验所证实,主要包括抗神经炎症反应、抗氧化应激损伤和抗缺氧缺血性神经损伤，可增加神经细胞的可塑性，从而减少海马神经元的凋亡。There is also a certain amount of sulfide in bamboo salt, with a typical stinky egg smell. Hydrogen sulfide has a variety of neuroprotective effects, which have been confirmed by a large number of experiments at home and abroad, mainly including anti-neuroinflammatory response, anti-oxidative stress injury and anti-hypoxic-ischemic nerve injury, which can increase the plasticity of nerve cells and reduce hippocampus Neuronal apoptosis.
此外，尤为值得一提的是，经过800℃以上的高温熔炼，盐的食用安全性大大提高，烤盐与原盐相比，重金属(铅、砷、汞)含量大幅度下降。同时，原盐中的其他有害成份(如有机的塑料微粒等)也被完全除去。In addition, it is particularly worth mentioning that after high temperature smelting above 800 ° C, the edible safety of the salt is greatly improved, and the content of heavy metals (lead, arsenic, and mercury) in the roasted salt is significantly reduced compared to the original salt. At the same time, other harmful components (such as organic plastic particles, etc.) in the raw salt are also completely removed.
表5不同试样的元素检测分析结果Table 5 Elemental analysis results of different samples
Figure PCTCN2019097358-appb-000005
Figure PCTCN2019097358-appb-000005
备注：磷元素检出限为20mg/kg；钼、锑、硒、锡、总砷元素检出限为0.01mg/kg；镉、*锗元素检出限为0.001mg/kg；铊元素检出限为0.0001mg/kg。Note: The detection limit of phosphorus is 20mg / kg; the detection limit of molybdenum, antimony, selenium, tin, and total arsenic is 0.01mg / kg; the detection limit of cadmium and * germanium is 0.001mg / kg; the detection of radon The limit is 0.0001 mg / kg.
实验2、植物功能盐及其对照品的显微结构和晶格结构分析 Experiment 2. Microstructure and Lattice Structure Analysis of Plant Functional Salts and Their References
2.1试样外观的显微镜图2.1 Microscope image of sample appearance
用体视显微镜(江南牌SE220)在目镜10倍、物镜5倍条件下拍摄得到实施中所得植物功能盐颗粒的试样外观如图8所示。A stereo microscope (Jiangnan Brand SE220) was used to capture the appearance of the sample of the plant functional salt particles obtained during the implementation under the conditions of 10 times the eyepiece and 5 times the objective lens, as shown in FIG. 8.
从图8中可以看到原盐和烤盐基本呈白色，颗粒表面较为光滑，外形规整。而松盐、竹盐和梅盐则呈现不同的颜色，晶体排列差异较大，更有光泽感。本发明制备的新品竹盐与仁山竹盐的色泽接近，但前者更加纯净，看上去晶莹剔透，而后者杂质较多。It can be seen from Figure 8 that the raw salt and roasted salt are basically white, the particle surface is relatively smooth, and the shape is regular. However, pine salt, bamboo salt, and plum salt show different colors, and the crystal arrangement differs greatly, giving them a more lustrous feel. The color of the new bamboo salt prepared by the present invention is similar to that of Renshan bamboo salt, but the former is more pure and looks crystal clear, while the latter has more impurities.
2.2扫描电镜的观察结果2.2 SEM observation results
利用德国Carl Zeiss Microscopy GmbH公司出品的GeminiSEM 300扫描电子显微镜对实施例的植物功能盐进行形态表征。分别将不同的试样颗粒置于电镜下观察，得到图9。原盐和烤盐表面光滑，无太多附着物。而经本发明所制得的植物功能盐表面呈现地瓜状，并且有许多小颗粒物质附着在颗粒表面。The GeminiSEM 300 scanning electron microscope produced by Carl Zeiss Microscopy GmbH of Germany was used to perform morphological characterization of the plant functional salts of the examples. The different sample particles were respectively observed under an electron microscope, and FIG. 9 was obtained. The surface of raw salt and roasted salt is smooth without much attachment. However, the surface of the plant functional salt prepared by the present invention is in the shape of a sweet potato, and many small particulate materials are attached to the particle surface.
2.3透射电镜的观察结果2.3 Observation results of transmission electron microscope
将不同的盐样悬浮于无水乙醇中(5％，w/v)，利用日本JEOL公司JEM-1230透射式电子显微镜(Transmission Electron Microscope，TEM)对实施例中所得植物功能盐颗粒进行形态表征。实施例4、5、6制得的松盐、竹、梅为代表的植物功能盐透射电镜图如图10所示，原盐形状较为规整，呈现氯化钠立方体规则结构，而植物功能盐无具体规则，且周围附着一些微小颗粒。Different salt samples were suspended in anhydrous ethanol (5%, w / v), and the morphological characterization of the plant functional salt particles obtained in the examples was performed using a JEM-1230 transmission electron microscope (TEM) from JEOL, Japan. . The transmission electron micrographs of the plant functional salts represented by the pine salt, bamboo, and plum prepared in Examples 4, 5, and 6 are shown in Fig. 10. The shape of the original salt is relatively regular, showing a regular structure of sodium chloride cubes, but the plant functional salt is not specific. Regular, and some small particles are attached around.
2.4 X射线衍射分析结果2.4 X-ray diffraction analysis results
将实施例4所得松盐、实施例5所得竹盐、实施例6所得梅盐及其对照试样研磨成粉末(过100目筛)，用德国Bruker公司生产的X射线衍射仪(Bruker D8Advance)进行分析。由于晶体是由原子规则排列成的晶胞组成，这些由不同原子散射的X射线相互干涉，在某些特殊方向上产生强X射线衍射，衍射线在空间分布的方位和强度与晶体结构密切相关，每种晶体所产生的衍射花样都反映出该晶体内部的原子分配规律。可对试样进行物相分析、晶胞参数测定及其衍射线强度分析。The pine salt obtained in Example 4, bamboo salt obtained in Example 5, plum salt obtained in Example 6 and its control sample were ground into powder (passed through a 100-mesh sieve), and an X-ray diffractometer (Bruker D8 Advance) produced by Bruker, Germany was used. Perform analysis. Since the crystal is composed of unit cells arranged regularly, these X-rays scattered by different atoms interfere with each other, resulting in strong X-ray diffraction in some special directions. The orientation and intensity of the diffraction line in spatial distribution are closely related to the crystal structure. The diffraction pattern produced by each crystal reflects the atomic distribution of the crystal. The sample can be analyzed for phase analysis, unit cell parameters, and diffraction line intensity analysis.
2.2.1晶体结构分析2.2.1 Crystal structure analysis
物相分析是X射线衍射在金属中用得最多的方面，分为定性分析和定量分析。前者把对材料测得的点阵平面间距及衍射强度与标准物相的衍射数据相比较，确定材料中存在的物相。精确的晶胞参数数据能够反映一种物质的不同样品间在结构上的细微差异，或者一种晶体的结构在外界物理化学因素作用下产生的微小变化。实施例中所得植物功能盐几种物相的晶胞参数如下：Phase analysis is the most commonly used aspect of X-ray diffraction in metals and is divided into qualitative and quantitative analysis. The former compares the lattice plane spacing and diffraction intensity measured on the material with the diffraction data of the standard phase to determine the phase present in the material. Accurate unit cell parameter data can reflect the slight differences in structure between different samples of a substance, or the small changes in the structure of a crystal caused by external physical and chemical factors. The unit cell parameters of several phases of the plant functional salt obtained in the examples are as follows:
表6不同试样中的氯化钠含量及晶格参数Table 6 Sodium chloride content and lattice parameters in different samples
Figure PCTCN2019097358-appb-000006
Figure PCTCN2019097358-appb-000006
表7不同试样中的氯化钾含量及晶格参数Table 7 Potassium chloride content and lattice parameters in different samples
Figure PCTCN2019097358-appb-000007
Figure PCTCN2019097358-appb-000007
表8盐的晶体大小Table 8 Salt crystal size
Figure PCTCN2019097358-appb-000008
Figure PCTCN2019097358-appb-000008
2.2.2X射线衍射谱图及模拟的晶体结构图2.2.2 X-ray diffraction spectrum and simulated crystal structure diagram
通过晶胞参数模拟出不同盐试样的晶胞图。从图11中可以看到原盐较为规整呈立方体形，而松盐两端较尖，竹盐较圆，梅盐则呈现出八面体的锥形，说明原盐与不同的植物提取物通过高温熔炼后，晶体结构发生了很大的变化。The unit cell parameters of different salt samples were simulated by unit cell parameters. From Figure 11, it can be seen that the raw salt is more regular and cubic, while the ends of the pine salt are sharper, the bamboo salt is more round, and the plum salt has an octahedral cone shape. The crystal structure has changed a lot.
实验3、植物功能盐及其对照品的感官性能测试 Experiment 3. Sensory performance test of plant functional salt and its reference
电子舌是模拟人的舌头对待测样品进行分析、识别和判断，用多元统计方法对得到的数据进行处理，快速地反映出样品整体的质量信息，实现对样品的识别和分类。是一种利用多传感阵列为基础，感知样品的整体特征响应信号，对样品进行模拟识别和定量定性分析的一种检测技术。主要由味觉传感器阵列、信号采集系统和模式识别系统3部分组成。The electronic tongue is to simulate the human tongue to analyze, identify and judge the sample to be tested, and use multivariate statistical methods to process the obtained data, quickly reflect the overall quality information of the sample, and realize the identification and classification of the sample. It is a detection technology that uses the multi-sensor array as the basis to sense the overall characteristic response signal of the sample, and performs analog identification and quantitative qualitative analysis on the sample. It is mainly composed of taste sensor array, signal acquisition system and pattern recognition system.
电子鼻又称气味扫描仪，是20世纪90年代发展起来的一种快速检测食品的新颖仪器。它以特定的传感器和模式识别系统快速提供被测样品的整体信息，指示样品的隐含特征。它是由选择性的电化学传感器阵列和适当的识别方法组成的仪器，能识别简单和复杂的气味，可得到与人的感官品评相一致的结果。Electronic nose, also known as odor scanner, is a novel instrument for rapid food detection developed in the 1990s. It uses specific sensors and pattern recognition systems to quickly provide overall information about the sample under test, indicating the underlying characteristics of the sample. It is an instrument composed of a selective array of electrochemical sensors and an appropriate identification method. It can identify simple and complex odors and can obtain results consistent with human sensory evaluation.
区分指数(Discrimination index,DI)是电子舌和电子鼻用来判定样品整体区分效果的一个衡量指标，其取值范围为[-100,+100]。-100,0表示其无法区分试样的味觉和嗅觉；+100表示其能够有效区分，其值越接近100，表示区分效果好。The discrimination index (DIcrimination index) is a measurement index used by the electronic tongue and electronic nose to determine the overall discrimination effect of the sample, and its value range is [-100, +100]. -100,0 indicates that it cannot distinguish the taste and smell of the sample; +100 indicates that it can effectively distinguish, and the closer the value is to 100, the better the discrimination effect.
3.1用电子舌得出的味觉性状3.1 Taste traits derived from electronic tongue
将实施例所得植物功能盐制成10％溶液，用电子舌(Smartongue，Isensogroup公司)检测其味觉差异。The plant functional salt obtained in the example was made into a 10% solution, and the difference in taste was detected with an electronic tongue (Smartongue, Isensogroup).
从图12中可以看出电子舌的DI值达到99.57，表明不同植物功能盐在味觉上存在显著差异，可以明显区分。主成分1(PC1)的贡献率为78.26％，远大于主成分2(PC2)；在横坐标上的距离越远，表示其差异性越大；实施例4所得松盐/实施例5所得竹盐和仁山竹盐的横坐标距离较近，表明二者滋味较接近，且都与原盐与实施例6所得梅盐有显著差异。It can be seen from Figure 12 that the DI value of the electronic tongue reaches 99.57, indicating that there is a significant difference in taste of different plant functional salts, which can be clearly distinguished. The contribution rate of the main component 1 (PC1) is 78.26%, which is much larger than that of the main component 2 (PC2); the longer the distance on the abscissa, the greater the difference; the pine salt obtained in Example 4 / bamboo obtained in Example 5 The abscissa distance between the salt and the mangosteen salt is close, indicating that the taste of the two is close, and both are significantly different from the original salt and the plum salt obtained in Example 6.
3.2用电子鼻得出的嗅觉性状3.2 Olfactory traits derived from electronic nose
电子鼻通过14个传感器收集不同样品的气味信号，再经软件处理将数据进行主成分分析，将实施例所得植物功能盐制成10％溶液，用电子鼻(iNose，Isensogroup公司)检测其味觉差异如图13所示：样品中占总信息量最多的两种主成分分别作为横坐标PC1和纵坐标PC2，主成分1(PC1)贡献率为99.80％远大于主成分2，因此两种样品数据在横坐标上的距离越远，表示其差异越大，图中植物功能盐之间横坐标非常接近，表明本发明得到的三种植物盐从嗅觉上看比较接近。而传统方法制备的竹盐(仁山竹盐)的气味与原盐较为接近。此外，根据表4原子能谱分析结果，竹盐表面发现有硫元素，因此可以解释其带有淡淡的硫化氢味，而其他盐的硫元素均为达到检测限。The electronic nose collects the odor signals of different samples through 14 sensors, and then analyzes the data by software for principal component analysis. The plant functional salt obtained in the example is made into a 10% solution, and the electronic nose (iNose, Isensogroup) is used to detect the difference in taste. As shown in Figure 13: the two principal components that account for the most information in the sample are abscissa PC1 and ordinate PC2, respectively. The contribution rate of principal component 1 (PC1) is 99.80%, which is much larger than that of principal component 2. Therefore, the two sample data The farther the distance on the abscissa is, the larger the difference is. The abscissas between the plant functional salts in the figure are very close, indicating that the three plant salts obtained by the present invention are closer from the olfactory perspective. The odor of bamboo salt (renshan bamboo salt) prepared by the traditional method is close to that of the original salt. In addition, according to the results of atomic spectrum analysis in Table 4, sulfur is found on the surface of bamboo salt, so it can be explained that it has a slight taste of hydrogen sulfide, while sulfur of other salts has reached the detection limit.
实验4、植物功能盐护肝作用的动物试验Experiment 4.Animal test of plant function salt to protect liver
4.1动物分组与实验设计4.1 Animal grouping and experimental design
4.1.1实验动物及分组4.1.1 Experimental animals and groups
取5周龄的SPF级C57BL/6健康雄性小鼠45只，适应性喂养一周后随机分成9组，实验设计及剂量配置如表9所示。除正常对照组(1 #)采用常规饲料外，其余8组均使用无盐基础饲料。给药阶段，根据世界卫生组织推荐的每人每日食盐摄入量5g及标准动物等效剂量折算法得出小鼠的口服剂量，除1 #组外，各灌胃样品含4％的NaCl，每日灌胃1次，连续21d。饲养环境温度为(22±2)℃，相对湿度为50～70％。 Forty-five 5-week-old SPF C57BL / 6 healthy male mice were randomly divided into 9 groups after one week of adaptive feeding. The experimental design and dosage configuration are shown in Table 9. Except for the normal control group (1 # ), which used conventional feed, the other 8 groups used salt-free basic feed. Administration phase, the oral dose results in mice according to the World Health Organization recommended daily salt intake of 5g per dose, and in standard animal equivalent conversion method, in addition to the group # 1, each sample was fed with 4% NaCl , Intragastric administration once a day for 21d. The breeding environment temperature is (22 ± 2) ° C, and the relative humidity is 50-70%.
表9动物试验分组及给药剂量Table 9 Animal test groups and doses
Figure PCTCN2019097358-appb-000009
Figure PCTCN2019097358-appb-000009
*为了确保试验数据的可比性，正常对照组2 #和模型组3 #膳食中的盐均为纯的NaCl * In order to ensure comparability test data, the normal control group and the model group # 2 # 3 diets are pure NaCl salt
4.1.2急性肝损伤造模4.1.2 Acute liver injury modeling
末次给药1h后，模型对照组和试验组(3 #～9 #)小鼠均腹腔注射0.1％的CCl 4橄榄油溶液，CCl 4剂量为10mL/kg.BW，正常对照组(1 #～2 #)给予等体积的橄榄油，禁食过夜。 After the last administration 1h, model control group and the experimental group (# 3 - # 9) mice were injected with 0.1% CCl 4 in olive oil solution, CCl 4 at a dose of 10mL / kg.BW, normal control group (# 1 ~ 2 # ) Give an equal volume of olive oil and fast overnight.
4.1.3试样采集及生化指标测定4.1.3 Sample collection and determination of biochemical indicators
在注射CCl 4橄榄油溶液16h后，脊椎脱臼处死小鼠，立刻摘眼球取血，同时解剖小鼠取肝脏。 Sixteen hours after the injection of the CCl 4 olive oil solution, the mice were sacrificed by spinal dislocation, the eyeballs were immediately removed for blood extraction, and the mice were dissected to take the liver.
血液样本，经3500r/min离心10min后，吸取上层血清用于血生化指标测定。取出完整肝脏后，剪取0.4g肝脏组织1份，放入装有PBS(pH＝7.4)的冻存管中匀浆，其余肝脏组织放入冻存管中，于-80℃冰箱中保存留用。将剪取的0.4g肝脏匀浆后置于超低温离心机中离心(4000r/min、20min)，取上清液，测定其丙二醛(MDA)、谷胱甘肽(GSH)的含量以及总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化酶(GSH-Px)的活性(考马斯亮蓝法)。After the blood sample was centrifuged at 3500r / min for 10min, the upper serum was collected for determination of blood biochemical indicators. After taking out the complete liver, cut a portion of 0.4g of liver tissue, put it into a cryopreserved tube filled with PBS (pH = 7.4), and homogenize the rest of the liver tissue in a cryopreserved tube. Store in a -80 ° C refrigerator . The cut 0.4g liver homogenate was centrifuged in an ultra-low temperature centrifuge (4000r / min, 20min), and the supernatant was taken to determine the content of malondialdehyde (MDA), glutathione (GSH), and total Activities of superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) (Coomassie blue method).
4.1.4肝组织的病理观察4.1.4 Pathological observation of liver tissue
取各组实验鼠相同部位的肝组织，冷生理盐水冲洗干净后用中性福尔马林固定，制作4μm的石蜡切片，行HE染色，在显微镜下观察其形态学的变化。Liver tissue from the same part of each group of rats was taken, washed with cold saline, fixed with neutral formalin, 4 μm paraffin sections were made, and HE stained, and the morphological changes were observed under a microscope.
4.1.5实验数据处理4.1.5 Experimental data processing
实验数据均采用均值±标准误差(x±s)表示，数据行方差齐性检验；组间数据比较采用单因素方差分析。使用SPSS19.0统计软件的单因素方差分析(ANOVA)中的Duncan检验分析显著性，P<0.05为差异显著，P<0.01为差异极显著。The experimental data were expressed as mean ± standard error (x ± s), and the data were tested for homogeneity of variance; the comparison of data between groups was performed by single factor analysis of variance. The Duncan test in the one-way analysis of variance (ANOVA) of SPSS19.0 statistical software was used to analyze the significance, P <0.05 was significant, and P <0.01 was extremely significant.
4.2结果与分析4.2 Results and analysis
4.2.1植物功能盐对实验小鼠体重的影响4.2.1 Effects of plant functional salts on body weight of experimental mice
在为期21d的试验周期内，各实验组小鼠的体质量随饲养时间延长，体质量均未表现出显著性差异(P>0.05)，观察各组小鼠皮毛光滑，临床活动表现均正常，健康状况良好，说明摄入不同的食盐样品对小鼠的临床表现无影响。试验周期中各组实验鼠的体质量情况详见表10。During the 21-day test period, the body weight of the mice in each experimental group increased with the feeding time, and the body weight did not show significant differences (P> 0.05). It was observed that the fur of the mice in each group was smooth and the clinical activities were normal. Good health status, indicating that intake of different salt samples had no effect on the clinical performance of mice. See Table 10 for the body weight of each group of experimental rats during the test cycle.
表10试验周期中各实验组小鼠的体质量比较Table 10 Comparison of body weight of mice in each experimental group during the test period
Figure PCTCN2019097358-appb-000010
Figure PCTCN2019097358-appb-000010
Figure PCTCN2019097358-appb-000011
Figure PCTCN2019097358-appb-000011
注：同一列两两之间若字母完全不同，则两者之间存在显著差异(P<0.05)；Note: If the letters in the same column are completely different, there is a significant difference between the two (P <0.05);
4.2.2植物功能盐对化学性肝损伤小鼠血清中转氨酶活性的影响4.2.2 Effects of plant functional salts on transaminase activity in serum of mice with chemical liver injury
从表11可见，模型组(3 #)小鼠血清谷氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)活性均极显著地高于正常对照组(2 #和1 #)(P<0.01)，表明造模成功。 As can be seen from Table 11, the serum glutamate aminotransferase (ALT) and aspartate aminotransferase (AST) activities of the model group (3 # ) mice were significantly higher than those of the normal control group (2 # and 1 # ) (P < 0.01), indicating successful modeling.
从表11可见，三种植物功能盐均表现出不同程度的降低血清转氨酶的活性，而原盐几乎无作用。与模型组相比，三种植物功能盐抑制ALT活性升高的作用极其显著(P<0.01)；在抑制AST活性升高上，松盐(P<0.01)比其余两种试样(P<0.05)更显著；松盐对CCl 4所致的急性肝损伤小鼠血清转氨酶急剧升高的抑制作用优于竹盐和梅盐。表明竹盐、松盐和梅盐具有一定的保肝护肝功效。然而，阳性药物对照组(联苯双酯)虽然具有最强的抑制ALT活性的功效，但却对AST无作用。市售的仁山紫竹盐也和原盐一样，未表现出对急性化学性肝损伤小鼠血清转氨酶活性的抑制作用。 It can be seen from Table 11 that the three plant functional salts all showed varying degrees of reducing serum transaminase activity, while the original salt had almost no effect. Compared with the model group, the effect of three plant functional salts on inhibiting the increase of ALT activity was extremely significant (P <0.01); in inhibiting the increase of AST activity, pine salt (P <0.01) was higher than the other two samples (P <0.01) 0.05) is more significant; the inhibitory effect of pine salt on serum liver aminotransferase in mice with acute liver injury induced by CCl 4 is better than that of bamboo salt and plum salt. It shows that bamboo salt, pine salt and plum salt have certain liver and liver protection effects. However, the positive drug control group (bifendate) had the strongest effect of inhibiting ALT activity, but had no effect on AST. The commercially available Renshan Zizhu salt, like the original salt, did not show an inhibitory effect on serum aminotransferase activity in mice with acute chemical liver injury.
表11各实验组小鼠血清中转氨酶活性的比较Table 11 Comparison of transaminase activities in serum of mice in each experimental group
Figure PCTCN2019097358-appb-000012
Figure PCTCN2019097358-appb-000012
注：同一列两两之间若字母完全不同，则两者之间存在显著差异(P<0.05)；Note: If the letters in the same column are completely different, there is a significant difference between the two (P <0.05);
模型对照组(3 #)与NaCl对照组(2 #)相比， #表示P<0.01； Compared with the model control group (3 # ) and the NaCl control group (2 # ), # means P <0.01;
各试验组(4 #～9 #)与模型对照组(3 #)相比，*表示P<0.01。 Compared with the model control group (3 # ) in each experimental group (4 # ~ 9 # ), * means P <0.01.
4.2.3植物功能盐对化学性肝损伤小鼠氧化应激指标的影响4.2.3 Effects of plant functional salts on oxidative stress indicators in mice with chemical liver injury
实验动物受到化学毒物攻击时，急性肝损伤的一个重要表征是MDA含量升高、GSH水平降低、SOD活力下降等，伴随着肝脏组织膜脂质的过氧化进而产生强烈的氧化应激反应。When experimental animals are attacked by chemical toxicants, an important characteristic of acute liver injury is an increase in MDA content, a decrease in GSH levels, and a decrease in SOD activity, etc., accompanied by the peroxidation of liver tissue membrane lipids and a strong oxidative stress response.
表12数据显示，同样摄入无盐饲料、并同时灌胃4％纯NaCl的二个组(2 #和3 #)用CCl 4造模以后，各项氧化应激指标都发生了显著变化，尤其是GSH和MDA水平的改变极其显著(P<0.01)，表明灌胃CCl 4后动物的肝脏确实受到了损伤。 The data in Table 12 show that the two groups (2 # and 3 # ) who also consumed salt-free feed and fed 4% pure NaCl at the same time made significant changes in oxidative stress indicators after modeling with CCl 4 . In particular, the changes in GSH and MDA levels were extremely significant (P <0.01), indicating that the livers of animals were indeed damaged after CCl 4 administration.
从表12可知，三种植物功能盐均可一定程度地升高受损肝脏的GSH含量，其中梅盐表现出极显著的提升作用；且均能降低受损肝脏的MDA含量，其中作用最强的是梅盐(P<0.01)，其次是仁山紫竹盐和松盐(P<0.01)；联苯双酯组也表现出升高GSH和降低MDA含量的作用，但效果均不如梅盐。在提升受损肝脏内源性抗氧化酶活性方面，三种植物功能盐均表现出一定作用，其中竹盐同时表现出对T-SOD活性的显著影响(P<0.05)。It can be known from Table 12 that all three kinds of plant functional salts can increase the GSH content of the damaged liver to a certain extent. Among them, the plum salt shows a very significant increase; and all of them can reduce the MDA content of the damaged liver. It was plum salt (P <0.01), followed by Renshan Zizhu salt and pine salt (P <0.01); the bifendate group also showed effects of increasing GSH and reducing MDA content, but the effects were not as good as those of plum salt. In terms of enhancing the endogenous antioxidant enzyme activity of the damaged liver, all three plant functional salts showed a certain effect, among which bamboo salt also showed a significant effect on T-SOD activity (P <0.05).
表12各实验组小鼠肝脏组织的抗氧化活性比较Table 12 Comparison of anti-oxidant activity in liver tissue of mice in each experimental group
Figure PCTCN2019097358-appb-000013
Figure PCTCN2019097358-appb-000013
注：同一列两两之间若字母完全不同，则两者之间存在显著差异(P<0.05)；Note: If the letters in the same column are completely different, there is a significant difference between the two (P <0.05);
模型对照组(3 #)与NaCl对照组(2 #)相比， #表示P<0.01； Compared with the model control group (3 # ) and the NaCl control group (2 # ), # means P <0.01;
各试验组(4 #～9 #)与模型对照组(3 #)相比，*表示P<0.01。 Compared with the model control group (3 # ) in each experimental group (4 # ~ 9 # ), * means P <0.01.
4.2.4实验动物肝脏损伤及其修复的组织病理学观察4.2.4 Histopathological observation of liver damage and repair in experimental animals
图14是植物功能盐对实验小鼠肝脏组织病理学的影响(×400)。1 #和2 #是正常小鼠的肝组织，可见肝细胞索排列整齐，细胞边界清晰，细胞核型正常，无明显异常；3 #NaCl模型组的肝细胞索排列紊乱，细胞核型发生明显变化，中央静脉中有大量细胞浸润；4 #联苯双酯组的肝细胞索排列较清晰，细胞核型正常，有少量空泡变性，表明联苯双酯滴丸对急性肝损伤有治疗作用；5 #仁山紫竹盐组的肝细胞索排列较清晰，细胞核型正常，有少量空泡变性和水样变性；6 #原盐对照组的肝细胞索排列不整齐，细胞核型发生明显变化，中央静脉中有大量细胞浸润；7 #竹盐试验组的肝细胞索排列较清晰，细胞边界清晰，细胞核型正常，有少量空泡变性；8 #梅盐试验组的肝细胞索排列清晰，细胞核型正常，有少量空泡变性；9 #松盐试验组的肝细胞索排列较清晰，细胞核型正常，有少量空泡变性，中央静脉中有少许细胞浸润。根据肝脏组织病理切片的形态学检查结果，可见竹盐、松盐、梅盐均对CCl 4所致的小鼠急性肝损伤有一定的防护作用。 Figure 14 is the effect of plant functional salts on the histopathology of the liver of experimental mice (× 400). # 1 and # 2 are mouse normal liver tissue, showing hepatic cords arranged in rows, clear cell borders, normal karyotype, no abnormalities; hepatic cords derangement 3 # NaCl model group karyotype change significantly, There is a large amount of cell infiltration in the central vein; hepatocyte cords in the 4 # biphenyldiester group are clearer, the karyotype is normal, and a small amount of vacuolar degeneration, indicating that bifendate drip pills have a therapeutic effect on acute liver injury; 5 # The arrangement of hepatocyte cords in Renshan Zizhu salt group was clear, the karyotype was normal, there was a small amount of vacuolar degeneration and watery degeneration; the hepatocyte cords of the # 6 original salt control group were irregularly arranged, the karyotype changed significantly, and a large number in the central vein Cell infiltration; hepatocyte cord arrangement in the 7 # bamboo salt test group was clear, cell boundaries were clear, the karyotype was normal, and there was a small amount of vacuole degeneration; hepatocyte cord in the 8 # plum salt test group was clearly arranged, and the karyotype was normal, with a small amount Vacuolar degeneration; The hepatocytes of the 9 # pine salt test group were arranged more clearly, the nuclei were normal, there was a small amount of vacuolar degeneration, and there was a little cell infiltration in the central vein. According to the morphological examination results of liver tissue pathological sections, it can be seen that bamboo salt, pine salt, and plum salt have certain protective effects on acute liver injury in mice induced by CCl 4 .
4.3结论4.3 Conclusion
我国是世界肝病第一大国，预测到2020年，慢性肝病人数将达4.5亿。肝病 高发的国情在很大程度上与食物来源、饮用水、饮食习惯和餐饮方式等有关。我国居民的食用盐主要来自海盐，日晒海盐(原盐)即海水的浓缩物，海洋中所有的污染物都有可能留存其中，带来一定的食品安全隐患。本研究结果显示，摄入原盐的实验鼠肝脏中GSH水平极显著地低于摄入同等剂量纯NaCl的组表明原盐中的确可能存在一些对肝脏不利的危害因素，造成了肝脏的氧化应激。最终结果显示，采用传统方法烤制的竹盐从竹筒中吸收到的有效成分是有限的，其护肝功效不及本发明提供的松、竹、梅盐。China is the world's largest liver disease country. It is predicted that by 2020, the number of chronic liver patients will reach 450 million. The national condition of the high incidence of liver disease is largely related to food sources, drinking water, eating habits, and dining styles. The edible salt of Chinese residents mainly comes from sea salt. The sun-sea salt (original salt) is the concentrate of seawater. All pollutants in the ocean may remain in it, bringing certain food safety risks. The results of this study show that the levels of GSH in the liver of experimental rats that ingested the raw salt were significantly lower than those in the same dose of pure NaCl, suggesting that there may indeed be some harmful factors in the liver that caused oxidative stress in the liver. The final results show that the traditional ingredients of the bamboo salt roasted by the traditional method are limited in the effective components absorbed from the bamboo tube, and their liver-protective effects are not as good as the pine, bamboo, and plum salts provided by the present invention.
实验5、植物功能盐的美白功效研究 Experiment 5. Study on the whitening effect of plant functional salt
5.1材料、试剂与仪器5.1 Materials, reagents and instruments
小鼠B16黑素瘤细胞，中国科学院上海细胞生物所；RPMI-1640培养液，美国Gibco公司；胎牛血清，美国ThermoFisher公司；胰蛋白酶、青霉素-链霉素溶液、磷酸缓冲液(PBS)，AR，杭州科易生物科技有限公司；左旋多巴(L-DOPA)、酪氨酸、酪氨酸酶(25KU，≥500u/mg)，AR，北京索莱宝科技有限公司；四甲基偶氮唑蓝(MTT，染色液质量分数为5g/L)，北京雷根生物技术有限公司；TritonX-100、-熊果苷、二甲基亚砜(DMSO)，AR，上海阿拉丁试剂有限公司。Mouse B16 melanoma cells, Shanghai Institute of Cell Biology, Chinese Academy of Sciences; RPMI-1640 medium, Gibco, USA; fetal bovine serum, ThermoFisher, USA; trypsin, penicillin-streptomycin solution, phosphate buffered saline (PBS), AR, Hangzhou Keyi Biotechnology Co., Ltd .; L-DOPA, tyrosine, tyrosinase (25KU, ≥500u / mg), AR, Beijing Solibao Technology Co., Ltd .; Tetramethyl coupling Azazole blue (MTT, 5g / L mass fraction of staining solution), Beijing Regen Biotechnology Co., Ltd .; TritonX-100, -arbutin, dimethyl sulfoxide (DMSO), AR, Shanghai Aladdin Reagent Co., Ltd. .
Eon酶标仪，美国BioTek公司；LDZX-50KBS立式高压灭菌锅，上海申安医疗器械厂；CP-ST50A型二氧化碳培养箱，长沙长锦科技有限公司；SW-CJ-F超净工作台，上海博迅实业有限公司；CKX41倒置显微镜，日本OLYMPUS光学工业株式会社；BDFACSCaLIBUR流式细胞仪，美国BD公司。Eon microplate reader, BioTek, USA; LDZX-50KBS vertical autoclave, Shanghai Shen'an Medical Equipment Factory; CP-ST50A carbon dioxide incubator, Changsha Changjin Technology Co., Ltd .; SW-CJ-F ultra-clean bench , Shanghai Boxun Industrial Co., Ltd .; CKX41 inverted microscope, Japan OLYMPUS Optical Industry Co., Ltd .; BDFACSCaLIBUR flow cytometer, American BD company.
5.2实验方法5.2 Experimental methods
5.2.1体外酪氨酸酶抑制率测定5.2.1 Determination of tyrosinase inhibition in vitro
以酪氨酸作为酪氨酸酶单酚酶活性测定的底物，以L-DOPA作为酪氨酸酶二酚酶活性测定的底物，参考Huang等的方法进行酪氨酸酶的活性测定。首先将原盐、烤盐、松盐、竹盐、梅盐用PBS溶液配成0.2％、1％、5％、10％质量分数的盐溶液，按表依次准确吸取不同浓度的样品溶液、0.1mol/L的pH6.8的PBS溶液和1.0mmol/L的酪氨酸或L-DOPA，充分混匀后放入25℃水浴锅孵育10min，然后加入相应的870u/mg酪氨酸酶溶液，10min后测定各反应溶液在475nm处的吸光值。抑制率按下式计算：Tyrosine was used as a substrate for tyrosinase monophenolase activity measurement, L-DOPA was used as a substrate for tyrosinase diphenolase activity measurement, and tyrosinase activity measurement was performed with reference to the method of Huang et al. First, the raw salt, roasted salt, pine salt, bamboo salt, and plum salt were prepared into a salt solution of 0.2%, 1%, 5%, and 10% by mass with PBS solution, and the sample solutions of different concentrations and 0.1mol were accurately sucked in order according to the table. / L pH6.8 PBS solution and 1.0mmol / L tyrosine or L-DOPA, mix thoroughly and put in a 25 ° C water bath and incubate for 10min, then add the corresponding 870u / mg tyrosinase solution for 10min The absorbance of each reaction solution at 475 nm was then measured. The suppression rate is calculated as follows:
酪氨酸酶单酚酶/二酚酶抑制率/％＝[1-(A3-A4)/(A1-A2)]×100Tyrosinase monophenolase / diphenolase inhibition rate /% = [1- (A3-A4) / (A1-A2)] × 100
表13体外酪氨酸酶活测定中的反应液组成(mL)Table 13 Composition of reaction solution in in vitro tyrosinase activity measurement (mL)
Figure PCTCN2019097358-appb-000014
Figure PCTCN2019097358-appb-000014
5.2.2 B16细胞增殖率的测定5.2.2 Determination of B16 cell proliferation rate
采用MTT法测定B16细胞增殖率，以原盐为阴性对照，-熊果苷和韩国竹盐作为阳性对照。选择对数生长期的B16细胞接入96孔板，贴壁后弃去原培养基，加入配好的含有0.25％、0.50％、0.75％和1％质量分数的原盐、松盐、竹盐、梅盐、-熊果苷和韩国竹盐的培养基，在37℃，5％CO 2条件下培养48h。在测定前4h取出，每孔加入20μL MTT(5mg/mL)溶液，放入CO 2培养箱中在37℃、5％CO 2环境中孵育4h，然后弃去培养基和MTT，向各孔中加入150μL DMSO，以溶解残留的MTT-甲臜结晶，振荡10min，以不加样品培养的细胞作为对照，PBS作为空白。用酶标仪测定各孔在570nm下的吸光值。细胞增殖率按下式计算： MTT method was used to determine the proliferation rate of B16 cells. The original salt was used as a negative control, and arbutin and Korean bamboo salt were used as positive controls. The B16 cells in the logarithmic growth phase were selected and inserted into a 96-well plate. After the adherence, the original culture medium was discarded, and the prepared raw salt, pine salt, bamboo salt, containing 0.25%, 0.50%, 0.75%, and 1% by mass were added. The culture medium of plum salt, -arbutin and Korean bamboo salt was cultured at 37 ° C and 5% CO 2 for 48 hours. Take out 4h before the measurement, add 20 μL of MTT (5mg / mL) solution to each well, put it in a CO 2 incubator and incubate for 4 h at 37 ° C, 5% CO 2 environment, then discard the medium and MTT, and put it in each well Add 150 μL of DMSO to dissolve the remaining MTT-formamidine crystals, shake for 10 min, and use the cells cultured without samples as a control and PBS as a blank. The absorbance of each well at 570 nm was measured with a microplate reader. The cell proliferation rate is calculated as follows:
细胞增殖率/％＝(OD 570(实验)－OD 570(空白))/(OD 570(对照)－OD 570(空白))×100 Cell proliferation rate /% = (OD 570 (experiment) -OD 570 (blank) ) / (OD 570 (control) -OD 570 (blank) ) × 100
5.2.3 B16细胞内酪氨酸酶活性的测定5.2.3 Determination of tyrosinase activity in B16 cells
采用多巴氧化法对B16细胞内酪氨酸酶活性进行测定。将B16细胞接入96孔板，贴壁后弃去原培养基，加入含有各样品的培养基进行培养48h。将作用时间已到的各组细胞倾去培养液，用PBS缓冲液洗2次，每孔加入1％质量分数的Triton-X100水溶液50μL，然后迅速放入-80℃冰箱内冻存30min，取出后在37℃下融化，使细胞完全破裂，每孔加入10μL L-DOPA溶液(10g/L)，以不加样品培养的细胞作为对照，PBS作为空白，37℃下反应2h，用酶标仪测定各孔在475nm处的OD值。按下式计算细胞内酪氨酸酶活性：Dopa oxidation method was used to determine the tyrosinase activity in B16 cells. B16 cells were inserted into a 96-well plate, the original medium was discarded after being adhered, and a medium containing each sample was added to culture for 48 h. The culture solution of each group of cells that has reached the action time is decanted, washed twice with PBS buffer, and 50 μL of a 1% by mass Triton-X100 aqueous solution is added to each well, and then quickly stored in a -80 ° C refrigerator for 30 minutes and removed. After thawing at 37 ° C, the cells were completely ruptured. 10 μL of L-DOPA solution (10g / L) was added to each well. Cells cultured without samples were used as a control, PBS was used as a blank, and the reaction was performed at 37 ° C for 2 hours. The OD value of each well at 475 nm was measured. Calculate intracellular tyrosinase activity as follows:
细胞内酪氨酸酶活性/％＝(OD 475(实验)－OD 475(空白))/(OD 475(对照)－OD 475(空白))/细胞增殖率×100 Intracellular tyrosinase activity /% = (OD 475 (experiment) -OD 475 (blank) ) / (OD 475 (control) -OD 475 (blank) ) / cell proliferation rate × 100
5.2.4 B16细胞迁移测定5.2.4 B16 cell migration assay
选择对数生长期的B16细胞，调整细胞浓度至4×10 5个/mL，取2mL加入6孔板，待细胞在底部覆盖60％左右时，弃去原培养基，取10μL移液枪头在每孔底部画直线(一次操作)，用PBS轻轻洗去滑落的细胞，再用0.5％质量分数的原盐、松盐、竹盐、梅盐、熊果苷配制的培养基继续培养48h，在显微镜下观察细胞的生长状态与迁移情况，以不加样品培养的细胞作为对照。 Select B16 cells in logarithmic growth phase, adjust the cell concentration to 4 × 10 5 cells / mL, add 2 mL to a 6-well plate, and when the cells cover about 60% at the bottom, discard the original medium and take 10 μL pipette tips Draw a straight line at the bottom of each well (one operation), gently wash off the slipped cells with PBS, and then continue to culture for 48 h with 0.5% mass fraction of the original salt, pine salt, bamboo salt, plum salt, and arbutin. The growth state and migration of the cells were observed under a microscope, and cells cultured without samples were used as controls.
5.2.5 B16细胞周期的测定5.2.5 Determination of B16 cell cycle
在6孔板中使B16细胞贴壁后，加入含有0.5％各试样的培养基进行培养，每组三个平行并设置空白对照组，分别培养48h。培养结束后收集细胞，用PBS清洗后去掉上清液，加入冷的70％乙醇溶液500μL，固定过夜，然后离心并吸取残留的乙醇。在细胞沉淀中加入RnaseA溶液100μL，重悬细胞，37℃水浴30min，再加入400μLPI染色液混匀，4℃避光孵育30min，转移至流式管中上机检测。After the B16 cells were adhered in a 6-well plate, a medium containing 0.5% of each sample was added for culture, and three groups of each group were set up in parallel and a blank control group was cultured for 48 hours. The cells were collected after the incubation, and the supernatant was removed after washing with PBS. 500 μL of a cold 70% ethanol solution was added and fixed overnight, then centrifuged and the remaining ethanol was aspirated. Add 100 μL of RnaseA solution to the cell pellet, resuspend the cells, incubate at 37 ° C for 30 minutes, add 400 μL PI staining solution and mix well, incubate at 4 ° C in the dark for 30 minutes, and transfer to a flow tube for detection on the machine.
5.2.6实验数据处理5.2.6 Experimental data processing
采用Origin9处理实验数据，计算结果以平均值±标准差(Mean±SD)表示，使 用SPSS 21单因素方差分析ANOVA中的Duncan进行显著性检验。The experimental data were processed using Origin9, and the calculation results were expressed as mean ± standard deviation (Mean ± SD). Significance tests were performed using Duncan in SPSS 21 one-way analysis of variance (ANOVA).
5.3结果分析5.3 Analysis of results
5.3.1植物盐对体外酪氨酸酶的抑制活性5.3.1 Inhibitory activity of plant salts on tyrosinase in vitro
当酪氨酸作为底物时，酪氨酸酶可将其催化为多巴并进一步生成黑色素，该过程主要是发挥了酪氨酸酶单酚酶活性。各种盐溶液对酪氨酸酶单酚酶活性的抑制作用如图15所示。由图15可知，原盐在0.2～10.0％浓度范围内均显示对酪氨酸酶无明显抑制作用。烤盐在5.0％浓度时对酪氨酸酶影响最大，但抑制率不高于10％。松盐与竹盐对酪氨酸酶活性影响显著，并且呈浓度效应关系，质量分数在0.2～5.0％时，随着盐浓度的增加，对酪氨酸酶的抑制率越大，5.0％时抑制率达到94％以上，10.0％时抑制率基本不变。相比于松盐和竹盐，梅盐在低浓度(0.2％、1％)时对酪氨酸酶抑制率较低，当浓度大于5.0％时抑制率迅速增加，10.0％的梅盐溶液对酪氨酸酶的抑制率为73.2％，效果略差于相同浓度的松盐、竹盐溶液。When tyrosine is used as a substrate, tyrosinase can catalyze it into dopa and further produce melanin. This process mainly exerts tyrosinase monophenolase activity. The inhibitory effect of various salt solutions on tyrosinase monophenolase activity is shown in FIG. 15. As can be seen from FIG. 15, the original salt showed no significant inhibitory effect on tyrosinase in the concentration range of 0.2 to 10.0%. Roasted salt had the greatest effect on tyrosinase at a concentration of 5.0%, but the inhibition rate was not higher than 10%. Pine salt and bamboo salt have a significant effect on tyrosinase activity and show a concentration-effect relationship. When the mass fraction is 0.2 to 5.0%, as the salt concentration increases, the greater the inhibition rate of tyrosinase, 5.0% The inhibition rate reached above 94%, and the inhibition rate remained basically unchanged at 10.0%. Compared with pine salt and bamboo salt, plum salt has a lower tyrosinase inhibition rate at low concentrations (0.2%, 1%), and the inhibition rate increases rapidly when the concentration is greater than 5.0%. The inhibition rate of tyrosinase was 73.2%, and the effect was slightly worse than that of pine salt and bamboo salt solutions of the same concentration.
当多巴作为底物时，酪氨酸酶可将其催化为多巴醌并进一步生成黑色素，该过程主要是发挥了酪氨酸酶二酚酶活性。各种盐溶液对酪氨酸酶二酚酶的抑制率如图16所示。由图可知，与对酪氨酸酶单酚酶抑制率相比，原盐和烤盐对酪氨酸酶二酚酶抑制率明显增强，并且有一定的浓度效应关系，但最大抑制率不超过30％。松盐、竹盐的酪氨酸酶二酚酶抑制与单酚酶抑制效果相似，高浓度(5.0％、10.0％)的梅盐溶液的二酚酶抑制率与相同浓度的松盐和竹盐溶液差别不大，5％时抑制率可达92.0％。植物盐与原盐相比，其钾、铁、镁、钙等金属元素含量较多，可能会对酪氨酸酶的蛋白质结构有一定影响，从而抑制了其活性。When dopa is used as a substrate, tyrosinase can catalyze it into dopaquinone and further produce melanin. This process mainly uses tyrosinase diphenolase activity. The inhibition rate of tyrosinase diphenolase by various salt solutions is shown in FIG. 16. It can be seen from the figure that compared with the inhibition rate of tyrosinase monophenolase, the inhibition rate of tyrosinase diphenolase by the original salt and roasted salt is obviously enhanced, and there is a certain concentration-effect relationship, but the maximum inhibition rate does not exceed 30 %. The tyrosinase diphenolase inhibition of pine salt and bamboo salt is similar to that of monophenolase. The high concentration (5.0%, 10.0%) of the plum salt solution has a diphenolase inhibition rate that is the same as that of pine salt and bamboo salt. The solution has little difference, and the inhibition rate can reach 92.0% at 5%. Compared with the original salt, plant salt has more potassium, iron, magnesium, calcium and other metal elements, which may have a certain effect on the protein structure of tyrosinase, thereby inhibiting its activity.
5.3.2植物功能盐对B16细胞增殖作用的影响5.3.2 Effects of plant functional salts on B16 cell proliferation
表14 B16细胞在各试样作用下的增殖率
Figure PCTCN2019097358-appb-000015
Table 14 Proliferation rate of B16 cells under the action of each sample
Figure PCTCN2019097358-appb-000015
Figure PCTCN2019097358-appb-000016
Figure PCTCN2019097358-appb-000016
注：不同字母表示在相同浓度下差异显著(p＜0.05)，n＝6。Note: Different letters indicate significant differences at the same concentration (p <0.05), n = 6.
黑色素是由黑色素细胞产生的，一般来说抑制黑色素细胞的增殖可以减少黑色素的生成。小鼠B16黑色素细胞在不同浓度的样品中的增殖率如表14所示。由 表14可知，0.25％浓度以上的各种盐溶液均会抑制B16细胞的生长，并且随着盐浓度的增加，细胞增殖率降低，在0.75％时各盐品组细胞增殖率差别不大，1％的盐溶液会严重影响细胞的生长，增殖率均低于10％。在低浓度(0.25％、0.5％)范围内，松盐、竹盐、梅盐以及韩国竹盐中的B16细胞的增殖率均不低于原盐，因此与原盐相比，植物盐对B16细胞没有明显的额外的细胞毒性，并且0.5％的松盐、竹盐、梅盐中细胞的增殖率远大于原盐，说明这几种植物盐对NaCl环境的B16细胞有一定的保护作用。阳性对照熊果苷组的细胞增殖率与浓度关系不大，均在70％左右。Melanin is produced by melanocytes. Generally speaking, inhibiting melanocyte proliferation can reduce melanin production. The proliferation rate of mouse B16 melanocytes in different concentrations of samples is shown in Table 14. It can be known from Table 14 that various salt solutions with a concentration of 0.25% or more can inhibit the growth of B16 cells, and with the increase of the salt concentration, the cell proliferation rate decreases. At 0.75%, the cell proliferation rates of the salt groups are not significantly different. 1% saline solution will seriously affect the cell growth, and the proliferation rate is less than 10%. In low concentrations (0.25%, 0.5%), the proliferation rate of B16 cells in pine salt, bamboo salt, plum salt, and Korean bamboo salt is not lower than that of the original salt. Therefore, compared with the original salt, plant salt has no effect on B16 cells. Obvious extra cytotoxicity, and the cell proliferation rate in 0.5% pine salt, bamboo salt, and plum salt is much greater than the original salt, indicating that these plant salts have a certain protective effect on B16 cells in the NaCl environment. The cell proliferation rate and concentration of the arbutin group in the positive control group had little relationship with each other, about 70%.
5.3.3植物盐对B16细胞内酪氨酸酶活性的影响5.3.3 Effect of plant salt on tyrosinase activity in B16 cells
酪氨酸酶是B16细胞合成黑色素的关键限速酶，其酶活是黑色素生成的重要影响因素，是衡量功能因子美白功效的重要指标。由表15可知，各试样对B16细胞的酪氨酸酶活性均有一定影响，其胞内酪氨酸酶活性均显著降低，且呈一定的浓度效应关系。其中与原盐相比，松盐、竹盐、韩国竹盐各组细胞酪氨酸酶活性均较高，说明这些植物盐对B16细胞酪氨酸酶的抑制作用不如原盐，而梅盐在每个浓度下细胞酪氨酸酶活性均小于原盐，说明梅盐可以显著降低B16细胞的酪氨酸酶活性，且效果优于各组受试物。阳性对照熊果苷组细胞酪氨酸酶活性均高于同浓度的盐样品组。Tyrosinase is a key rate-limiting enzyme for melanin synthesis in B16 cells. Its enzyme activity is an important factor affecting melanin production and an important indicator for measuring the whitening efficacy of functional factors. It can be known from Table 15 that each sample has a certain effect on the tyrosinase activity of B16 cells, and the intracellular tyrosinase activity is significantly reduced, and there is a certain concentration-effect relationship. Compared with the original salt, pine salt, bamboo salt, and Korean bamboo salt have higher tyrosinase activity in each group of cells, indicating that these plant salts have less inhibitory effect on tyrosinase in B16 cells than the original salt. The tyrosinase activity of the cells was lower than that of the original salt at the concentration, indicating that plum salt can significantly reduce the tyrosinase activity of B16 cells, and the effect is better than the test substances in each group. The tyrosinase activity of cells in the positive control arbutin group was higher than that in the salt sample group of the same concentration.
表15 B16细胞在各试样作用下的酪氨酸酶活性
Figure PCTCN2019097358-appb-000017
Table 15 Tyrosinase activity of B16 cells under the effect of each sample
Figure PCTCN2019097358-appb-000017
Figure PCTCN2019097358-appb-000018
Figure PCTCN2019097358-appb-000018
注：不同字母表示在相同浓度下差异显著(p＜0.05)，n＝6。Note: Different letters indicate significant differences at the same concentration (p <0.05), n = 6.
5.3.4植物盐对B16细胞迁移的作用5.3.4 Effect of plant salt on B16 cell migration
肿瘤细胞的重要特征是可以在增殖过程中进行扩散转移，可以通过其迁移度来判断其扩散能力。由图17可知，正常组(a)的B16细胞在培养48h后划线区域基本愈合，说明其迁移能力很强。当受试物质量分数为0.5％时，原盐(b)作用的B16细胞在划线区只有部分细胞进入，说明原盐对细胞迁移有一定的抑制作用。松盐(d)、竹盐(e)、梅盐(f)作用的细胞中，划线区基本没有细胞进入， 说明这三种植物盐可以显著抑制B16细胞的转移，且效果强于原盐。熊果苷(c)对B16细胞的迁移率也有一定抑制作用，但其效果不如植物盐。An important feature of tumor cells is that they can spread and metastasize during proliferation, and their ability to spread can be judged by their degree of migration. As can be seen from FIG. 17, the B16 cells in the normal group (a) basically healed after 48 h of culture, indicating that their migration ability is very strong. When the mass fraction of the test substance was 0.5%, B16 cells treated with the original salt (b) entered only part of the cells in the underlined area, indicating that the original salt had a certain inhibitory effect on cell migration. In the cells affected by pine salt (d), bamboo salt (e), and plum salt (f), there was almost no cell entry in the underlined area, indicating that these three plant salts can significantly inhibit the B16 cell metastasis and have stronger effects than the original salt. Arbutin (c) also has a certain inhibitory effect on the mobility of B16 cells, but its effect is not as good as that of plant salts.
5.3.5植物盐对B16细胞周期的影响5.3.5 Effect of plant salt on B16 cell cycle
细胞周期是指由细胞分裂结束到下一次细胞分裂结束所经历的过程，共分为四个阶段，G0/G1期(DNA合成前期)、S期(DNA复制期)、G2期(DNA复制完成到有丝分裂开始前)、M期(细胞分裂开始到结束)。不同周期的细胞中DNA含量不同，利用荧光染料PI将细胞DNA染色，并在流式细胞仪中检测荧光强度，即可研究细胞在不同周期阶段的分布，了解细胞的增殖情况。由表16可知，原盐与熊果苷组与正常组细胞在G0/G1期和S期分布无显著差异，而松盐、竹盐与梅盐组S期与G2/M期分布比例明显增加，G0/G1期分布较少。S期与G2/M的总和代表了细胞在DNA复制和有丝分裂过程的比例，可以表示细胞的增殖能力，结果显示松盐、竹盐组细胞增殖能力较好，梅盐组次之，其余各组增殖能力均无明显变化。MTT结果显示各受试物对B16细胞群的增殖有一定抑制作用，而在细胞周期研究发现存活的细胞增殖能力较好，推测受试物处理可能会引起部分细胞的死亡，而对一部分耐受的细胞具有促进增殖的效果。The cell cycle refers to the process from the end of cell division to the end of the next cell division. It is divided into four stages, G0 / G1 (pre-DNA synthesis), S (DNA replication), and G2 (DNA replication completed). To the beginning of mitosis), M phase (beginning to the end of cell division). The DNA content of cells in different cycles is different. Using the fluorescent dye PI to stain the cell DNA and detecting the fluorescence intensity in a flow cytometer, you can study the distribution of cells in different cycle stages and understand the cell proliferation. It can be seen from Table 16 that the distribution of cells in the original salt, arbutin group and normal group was not significantly different in the G0 / G1 and S phases, while the distribution ratios of the S and G2 / M phases in the pine salt, bamboo salt, and plum salt groups increased significantly. G0 / G1 phase is less distributed. The sum of the S phase and G2 / M represents the proportion of cells in the process of DNA replication and mitosis, which can indicate the cell's proliferation ability. The results show that the pine salt and bamboo salt groups have better cell proliferation ability, followed by the plum salt group, and the rest of the groups. There were no significant changes in proliferation ability. MTT results show that each test substance has a certain inhibitory effect on the proliferation of the B16 cell population, and cell cycle studies have found that the viability of surviving cells is better. It is speculated that the treatment of the test substance may cause some cell death and be resistant to some The cells have the effect of promoting proliferation.
表16 0.5％质量分数各试样对B16细胞周期的影响Table 16 Effect of 0.5% mass fraction of each sample on B16 cell cycle
Figure PCTCN2019097358-appb-000019
Figure PCTCN2019097358-appb-000019
注：不同字母表示在相同细胞周期下差异显著(p＜0.05)，n＝3。Note: Different letters indicate significant differences under the same cell cycle (p <0.05), n = 3.
5.4结论5.4 Conclusion
本发明提供的松、竹、梅盐作为三种代表性的植物功能盐，对体外酪氨酸酶的单酚酶活性及二酚酶活性均表现出浓度依赖的抑制作用，当试样浓度为10％时，酪氨酸酶单酚酶抑制率分别达到97.8％、94.2％、73.3％；浓度为5％时，二酚酶抑制率分别达到99.9％、93.0％、92.0％。植物功能盐对B16细胞增殖有一定的抑制作用，但与原盐相比无明显的细胞毒性。梅盐可以显著降低B16细胞内酪氨酸酶活性，0.75％梅盐培养基中的B16细胞酪氨酸酶活性仅有6.07％。同时，植物功能盐可以显著抑制B16细胞的横向迁移，但并不通过影响细胞周期来抑制其增殖。植物功能盐作为体外及胞内酪氨酸酶无机抑制剂，其抑制效果良好，可以考虑作为果蔬保鲜剂和人体的美白功能成分。As three representative plant functional salts, the pine, bamboo, and plum salts provided by the present invention exhibit concentration-dependent inhibition on monophenolase activity and diphenolase activity of tyrosinase in vitro. When the sample concentration is At 10%, the inhibition rates of tyrosinase monophenolase reached 97.8%, 94.2%, and 73.3%, respectively; at a concentration of 5%, the inhibition rates of diphenolase reached 99.9%, 93.0%, and 92.0%, respectively. Plant functional salt has a certain inhibitory effect on B16 cell proliferation, but it has no obvious cytotoxicity compared to the original salt. Plum salt can significantly reduce tyrosinase activity in B16 cells, and B16 cell tyrosinase activity in 0.75% plum salt medium is only 6.07%. At the same time, plant functional salts can significantly inhibit the lateral migration of B16 cells, but do not inhibit their proliferation by affecting the cell cycle. Plant functional salts, as inorganic inhibitors of tyrosinase in vitro and in vivo, have good inhibitory effects, and can be considered as fruit and vegetable fresheners and whitening functional components of the human body.
实验6、植物功能盐预防高血压的动物试验研究 Experiment 6. Animal experimental study of plant functional salt to prevent hypertension
6.1实验动物来源及分组情况6.1 Source and grouping of experimental animals
实验动物：雄性大鼠SD大鼠，13～14周龄，体重200g左右。适应性喂养一周后随机分成8组，即摄食正常盐饲料对照组、灌胃正常盐水无盐饲料对照组、高盐模型组、实验组(松、竹、梅植物功能盐)、阳性对照组(非洛地平缓释片)，每组4只，实验设计及剂量配置如表17所示。实验期间，单笼饲养，饲养温度为25℃，湿度30％，每周观察临床表现。给药阶段，大鼠每天高盐摄入量为4000mg/kg(相当于含盐量8％日粮)，在自由饮服纯净水的前提下，将各样品和对照试样配置成相应浓度的溶液，每天给大鼠经口灌喂2mL/100g的竹盐水、松盐水和梅盐水等。除正常对照组(1 #)采用常规饲料外，其余8组均使用无盐基础饲料。 Experimental animals: male rats SD rats, 13-14 weeks old, weighing about 200g. After one week of adaptive feeding, they were randomly divided into 8 groups, namely the control group fed with normal salt, the control group fed with normal saline without salt, the high salt model group, the experimental group (functional salt of pine, bamboo, and plum plants), and the positive control group Felodipine sustained-release tablets), 4 rats in each group, and the experimental design and dosage configuration are shown in Table 17. During the experiment, the animals were raised in a single cage at a temperature of 25 ° C and a humidity of 30%. Clinical manifestations were observed weekly. During the administration period, the rats' daily high salt intake was 4000mg / kg (equivalent to 8% dietary salt). On the premise of free drinking pure water, the samples and control samples were configured to the corresponding concentration. The solution was orally administered to rats daily with 2 mL / 100 g of bamboo saline, pine saline, and plum saline. Except for the normal control group (1 # ), which used conventional feed, the other 8 groups used salt-free basic feed.
表17动物试验分组及给药剂量Table 17 Animal test groups and doses
Figure PCTCN2019097358-appb-000020
Figure PCTCN2019097358-appb-000020
*为了确保试验数据的可比性，自由饮食含0.3％食盐的常规饲料(组1 #)和经口灌喂等质量的原盐水溶液、同时摄食无盐饲料的对照组(组2 #)，以比较灌喂方式摄入盐与饲料中摄入盐对大鼠的影响。 * In order to ensure the comparability of the test data, a free diet containing 0.3% common salt (group 1 # ) and a control group (group 2 # ) fed with an equivalent amount of raw saline solution while ingesting salt-free feed were compared. Effects of salt ingestion and salt ingestion on rats.
**高盐饲料组添加量参考文献饲料中加入8％盐折算为灌胃浓度。** References to the amount of high-salt feed group added 8% salt in the feed was converted to the gavage concentration.
6.2试验情况6.2 Test situation
每4天测一次血压与体重，试验持续27天。最后一次灌胃后，禁食不禁水，置于代谢笼24h并收集尿液。结束后腹腔注射麻药，心脏取血后处死大鼠。Blood pressure and weight were measured every 4 days, and the test lasted 27 days. After the last gastric lavage, fasting and water were not allowed, placed in a metabolic cage for 24 hours and urine was collected. After the end, anesthetic was injected intraperitoneally, and the rats were sacrificed after blood was drawn from the heart.
6.3试验结果6.3 Test results
6.3.1实验大鼠的体重变化6.3.1 Changes in body weight of experimental rats
实验24天内各组动物体重无显著差异，灌胃、高盐摄入均对体重无显著影响。There was no significant difference in body weight between animals in each group within 24 days of the experiment, and no significant effect on body weight was observed after gavage or high salt intake.
表18体重变化表Table 18 Weight Change Table
Figure PCTCN2019097358-appb-000021
Figure PCTCN2019097358-appb-000021
常规饲料(组1)与灌胃常规原盐(组2)相比， Δ表示p<0.05， ΔΔ表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), Δ indicates p <0.05, ΔΔ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
6.3.2实验大鼠收缩压的变化6.3.2 Changes in systolic blood pressure in experimental rats
灌胃高剂量盐到第20天时高盐实验组3与常规原盐组2已出现显著性差异，说明通过口服高剂量的盐会导致高血压，实验造模成功。从实验组来看，到第24天时，通过实施例所得的松、竹、梅植物功能盐组均与原盐组形成显著性差异，说明摄食本实施例制备的植物功能盐具有预防高血压效果。而值得注意的是，植物盐中梅盐降压效果最为明显，与低盐摄入无显著性差异，有可能是梅盐特殊的理化结构及元素含量有关。At the 20th day of high-dose salt gavage, the high-salt test group 3 and the conventional raw salt group 2 had a significant difference, indicating that oral high-dose salt can lead to hypertension, and the experimental modeling was successful. From the experimental group, by the 24th day, the functional salt groups of the pine, bamboo, and plum plants obtained in the examples all had significant differences from the original salt groups, indicating that ingestion of the plant functional salts prepared in this example had the effect of preventing hypertension. It is worth noting that plum salt has the most obvious antihypertensive effect in plant salt, and there is no significant difference with low salt intake. It may be due to the special physicochemical structure and element content of plum salt.
表19大鼠血压变化值Table 19 Changes in blood pressure in rats
Figure PCTCN2019097358-appb-000022
Figure PCTCN2019097358-appb-000022
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
6.3.3实验大鼠24h的尿量变化6.3.3 24h urine volume change in experimental rats
常规饲料和常规原盐组无显著性差异，说明灌喂对尿量无明显影响。因摄食高浓度的盐水导致饮水量增加，因而高盐组尿量显著性高于正常食盐组。但在植物功能盐中，竹盐的尿量明显高于其他组，和原盐组形成显著性差异，说明竹盐具有显著利尿作用。值得注意的是竹盐组的尿量高于阳性对照，非洛地平是一种选择性钙离子拮抗药，主要抑制小动脉平滑肌细胞外钙的内流，有促尿钠排泄和利尿作用。结果表明竹盐的利尿效果尤为显著。There was no significant difference between the conventional feed and the conventional raw salt group, indicating that feeding had no significant effect on urine output. Due to the increase in drinking water caused by the consumption of high-concentration saline, the urine output of the high-salt group was significantly higher than that of the normal salt group. However, in the plant functional salt, the urine volume of bamboo salt was significantly higher than other groups, and a significant difference was formed with the original salt group, indicating that bamboo salt has a significant diuretic effect. It is worth noting that the urine volume of the bamboo salt group is higher than that of the positive control. Felodipine is a selective calcium antagonist that mainly inhibits the influx of extracellular calcium from small arterial smooth muscle, and has urinary sodium excretion and diuretic effects. The results show that the diuretic effect of bamboo salt is particularly significant.
表20大鼠尿量值Table 20 Rat urine output values
Figure PCTCN2019097358-appb-000023
Figure PCTCN2019097358-appb-000023
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
6.3.4实验大鼠血清生化指标的分析结果6.3.4 Analysis results of serum biochemical parameters of experimental rats
(1)肝功能(1) Liver function
丙氨酸氨基转移酶(ALT)，是一种参与人体蛋白质新陈代谢的酶，是肝脏病变程度的重要指标。ALT可以加快蛋白质氨基酸在体内的转化，在肝脏细胞中含量最多。当组织器官病变时，就会把其中的ALT释放到血液中，使血清ALT含量增加。与正常大鼠相比，高盐组大鼠血清中ALT显著增加，说明肝脏受到一定损伤。Alanine aminotransferase (ALT), an enzyme involved in the metabolism of human proteins, is an important indicator of the degree of liver disease. ALT can accelerate the conversion of protein amino acids in the body, and it is the most abundant in liver cells. When tissues and organs are diseased, the ALT in them will be released into the blood, which will increase the serum ALT content. Compared with normal rats, ALT in the serum of the high-salt group was significantly increased, indicating that the liver was damaged.
天门冬氨酸氨基转移酶(AST)有助于了解心肌、肝、肾组织的损伤程度。AST存在于人体各种组织中，以心肌含量最丰富，其次是肝脏。当心和肝细胞出现坏死时，m-AST从线粒体中释放出来，使得血清中的AST增高。高盐组大鼠血清中AST显著增加，说明摄食高浓度的盐后肝脏受到一定影响。Aspartate aminotransferase (AST) helps to understand the degree of damage to myocardium, liver and kidney tissue. AST exists in various tissues of the human body, with the heart muscle being the most abundant, followed by the liver. When necrosis of the heart and liver cells occurs, m-AST is released from the mitochondria, which increases the AST in the serum. The AST in the serum of rats in the high-salt group was significantly increased, indicating that the liver was affected to some extent after ingesting high concentrations of salt.
碱性磷酸酶(ALP)广泛分布于人体肝脏、骨骼、肠、肾和胎盘等组织，经肝脏向胆外排出，主要用于骨骼、肝胆系统疾病的诊断和鉴别诊断，ALP病理性的增高与骨骼、肝胆疾病紧密相关。各组ALP含量无显著性差异，说明在一个月内摄食高盐饮食对ALP影响不太明显。Alkaline phosphatase (ALP) is widely distributed in human liver, bones, intestines, kidneys and placenta. It is excreted by the liver to the gallbladder. It is mainly used for the diagnosis and differential diagnosis of bone, hepatobiliary system diseases. Bone, liver and gallbladder diseases are closely related. There was no significant difference in ALP content between the groups, which indicated that the effect of high-salt diet on ALP in one month was not obvious.
表21肝功能指标Table 21 liver function indicators
Figure PCTCN2019097358-appb-000024
Figure PCTCN2019097358-appb-000024
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
(2)肾功能(2) Renal function
尿素氮(BUN)是血浆中除蛋白质以外的一种含氮化合物，它从肾小球滤过而排出体外。在肾功能不全失代偿时，BUN将升高。临床将其作为判断肾小球滤过功能的指标；肌酐(CRea)是是肌肉组织在体内代谢的产物，主要由肾小球滤过全部排出体外，当肾功能不全时，肌酐会在体内蓄积成为对人体有害的毒素；当血中尿酸全部从肾小球滤过，肾处理尿功能异常会使得血液尿酸(UA)含量增加。从表中可以看到在一月以内各组指标均无显著性差异，有可能是高盐饮食时间太短。Urea nitrogen (BUN) is a nitrogen-containing compound other than protein in the plasma. It is filtered from the glomerulus and excreted. When renal insufficiency is decompensated, BUN will increase. It is used clinically as an index for judging glomerular filtration function; creatinine (CRea) is a metabolic product of muscle tissue in the body, which is mainly excreted by glomerular filtration. When renal insufficiency, creatinine will accumulate in the body It becomes a harmful toxin to the human body; when all the uric acid in the blood is filtered from the glomeruli, abnormal urine processing by the kidneys will increase the blood uric acid (UA) content. It can be seen from the table that there is no significant difference between the indicators in each group within one month, and it may be that the high salt diet time is too short.
表22肾功能指标Table 22 renal function indicators
Figure PCTCN2019097358-appb-000025
Figure PCTCN2019097358-appb-000025
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
(3)心肌功能(3) Myocardial function
骨骼肌、心肌、平滑肌中肌酸激酶(CK)含量为多，主要存在于细胞质和线粒体中，是一个与细胞内能量运转、肌肉收缩、ATP再生有直接关系的重要激酶。肌酸激酶活性测定可以用于骨骼肌疾病及心肌疾病的诊断。病理性增高：心肌梗死、病毒性心肌炎、皮肌炎、肌营养不良、心包炎、脑血管意外破裂等。从表中可以看到摄食高浓度的盐对CK含量有显著性影响，一个月多的高盐饮食可能对心肌功能有一定影响。但值得注意的是松盐CK含量显著性低于高盐组，说明松盐在预防心肌损伤上具有潜力。Skeletal muscle, cardiac muscle, and smooth muscle are rich in creatine kinase (CK), which are mainly present in the cytoplasm and mitochondria, and are an important kinase that is directly related to intracellular energy movement, muscle contraction, and ATP regeneration. The measurement of creatine kinase activity can be used for the diagnosis of skeletal muscle disease and myocardial disease. Pathological increase: myocardial infarction, viral myocarditis, dermatomyositis, muscular dystrophy, pericarditis, accidental rupture of cerebral blood vessels, etc. It can be seen from the table that high salt concentration has a significant effect on CK content, and a high salt diet for more than a month may have a certain effect on myocardial function. However, it is worth noting that the CK content of pine salt is significantly lower than that of the high salt group, indicating that pine salt has the potential to prevent myocardial damage.
乳酸脱氢酶(LDH)几乎存在于所有组织中，以心、骨骼肌和肾脏最丰富，可用于心肌疾病的诊断。乳酸脱氢酶增高：主要见于心肌梗死、肝炎、恶性肿瘤、肺梗塞、白血病、溶血性贫血、肾脏疾病、进行性肌萎缩等。高盐饮食CK、LDH含量均低于正常盐饮食组，进一步说明了，高盐饮食对心肌功能的损伤。Lactate dehydrogenase (LDH) is found in almost all tissues and is most abundant in heart, skeletal muscle, and kidney, and can be used for the diagnosis of myocardial disease. Increased lactate dehydrogenase: mainly seen in myocardial infarction, hepatitis, malignant tumors, pulmonary infarction, leukemia, hemolytic anemia, kidney disease, progressive muscle atrophy, etc. The contents of CK and LDH in the high-salt diet were lower than those in the normal salt diet group, which further explained that the high-salt diet had damage to myocardial function.
表23心肌功能指标Table 23 Myocardial function indicators
Figure PCTCN2019097358-appb-000026
Figure PCTCN2019097358-appb-000026
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
(4)血清离子含量(4) Serum ion content
一个月的实验结果表明血清中各离子浓度差异不大。The results of one month's experiment showed that there was not much difference in the concentration of each ion in the serum.
表24血清离子含量Table 24 Serum ion content
Figure PCTCN2019097358-appb-000027
Figure PCTCN2019097358-appb-000027
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
6.3.5实验大鼠的尿液指标分析结果6.3.5 Analysis results of urine parameters of experimental rats
(1)总排泄量(1) Total excretion
尿肌酐(CRea)主要来自血液，经由肾小球过滤后随尿液排除体外，肾小管基本不吸收且排出很少。当肾脏出现问题时尿肌酐含量会发生变化。正常尿液中含少量小分子蛋白(U-TP)，当尿中蛋白增加便形成蛋白尿，蛋白尿是肾脏病的常见表现，全身性疾病亦可出现蛋白尿。尿微量白蛋白(U-ALB)是重要的血浆蛋白质之一，在正常情况下，白蛋白的分子量大，不能越过肾小球基膜。疾病时肾小球基膜受到损害致使通透性发生改变，导致白蛋白的排出从而进入尿液中，使得尿白蛋白浓度持续升高。从表中可以看到一个月高盐饮食对SD大鼠U-TP和 U-ALB含量出现显著性差异，说明肾功能有所损伤，但从血清肾脏指标及尿液中CRea含量无显著性变化，说明一个月高盐饮食影响程度还不强，若长期性高盐饮食对肾脏会造成损伤。值得注意的是松盐U-TP水平显著性低于高盐组，与阳性对照相当，说明松盐也具有保护肾功能的效果。Urinary creatinine (CRea) is mainly from the blood. After filtering through the glomerulus, it is eliminated with the urine. The renal tubules are basically not absorbed and excreted rarely. Urine creatinine content changes when kidney problems occur. Normal urine contains a small amount of small molecule protein (U-TP). When protein in the urine increases, proteinuria is formed. Proteinuria is a common manifestation of kidney disease, and proteinuria can also occur in systemic diseases. Urinary microalbumin (U-ALB) is one of the important plasma proteins. Under normal circumstances, albumin has a large molecular weight and cannot cross the glomerular basement membrane. Damage to the glomerular basement membrane during disease results in a change in permeability, leading to the excretion of albumin into the urine, causing the urine albumin concentration to continue to increase. It can be seen from the table that the one-month high-salt diet has significant differences in U-TP and U-ALB contents in SD rats, indicating that renal function is impaired, but there is no significant change in serum kidney indexes and CRea content in urine. This indicates that the effect of a high-salt diet for one month is not strong. If a long-term high-salt diet will cause kidney damage. It is worth noting that the U-TP level of pine salt is significantly lower than that of the high salt group, which is equivalent to the positive control, indicating that pine salt also has the effect of protecting renal function.
表25尿液生化指标Table 25 Urine biochemical indicators
Figure PCTCN2019097358-appb-000028
Figure PCTCN2019097358-appb-000028
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
(2)尿液钙磷镁钠氯钾离子排泄量(2) excretion of urine calcium, phosphate, magnesium, sodium, potassium and potassium
除了Na和Cl离子含量增加外，高盐组内K、Ca、P含量明显增加，其中竹盐中P元素较为明显。在植物盐元素分析中，原盐和一烤盐P元素均无检出，而松竹梅盐检出分别为71.8、45.3、424mg/kg。从植物功能盐理化指标中可以看到其元素含量明显多于原盐及烤盐，但其尿液排泄量却无显著性差异，说明植物功能盐中丰富的元素被体内吸收。In addition to the increase in Na and Cl ion content, the K, Ca, and P contents increased significantly in the high-salt group, and the P element in bamboo salt was more pronounced. In the analysis of plant salt elements, neither the original salt nor the P element of a roasted salt was detected, while the salt of Songzhumei was 71.8, 45.3, and 424 mg / kg, respectively. From the physical and chemical indicators of plant functional salt, it can be seen that its element content is significantly more than that of raw salt and roasted salt, but its urine excretion is not significantly different, indicating that the rich elements in plant functional salt are absorbed by the body.
表26尿液离子排泄量Table 26 Urine ion excretion
Figure PCTCN2019097358-appb-000029
Figure PCTCN2019097358-appb-000029
常规饲料(组1)与灌胃常规原盐(组2)相比， △表示p<0.05， △△表示p<0.01； Compared with the conventional raw salt (group 2) given by oral administration (group 1), △ indicates p <0.05, △△ indicates p <0.01;
高盐试验组(组3)与原盐正常组(组2)相比， #表示p<0.05， ##表示p<0.01； In the high salt test group (group 3), compared with the normal salt group (group 2), # means p <0.05, ## means p <0.01;
各高盐试验组(组4～8)与高盐试验组(组3)相比， *表示p<0.05， **表示p<0.01。 Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8) * indicates p <0.05, and ** indicates p <0.01.
6.4结论6.4 Conclusion
普通盐和烤盐在预防SD大鼠高血压试验中无显著性差异，说明同样以本发明制备的无添加植物提取物的烤盐并无特殊的生物学功效。There is no significant difference between ordinary salt and roasted salt in the prevention of hypertension in SD rats, which indicates that roasted salt without added plant extract prepared by the present invention has no special biological effect.
经过一个月的高盐饮食，雄性SD大鼠摄食原盐后收缩压明显上升，并表现出明显的肝、肾、心肌功能损伤；而摄食本发明的植物功能盐的实验动物血压则明显低于原盐，表明其具有替代普通食用盐、防治高血压的潜力。After a month of high-salt diet, the systolic blood pressure of male SD rats significantly increased after taking raw salt, and showed obvious liver, kidney, and myocardial function damage; while the blood pressure of experimental animals ingesting the plant functional salt of the present invention was significantly lower than the original salt , Indicating that it has the potential to replace ordinary edible salt and prevent hypertension.
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。All documents mentioned in the present invention are incorporated by reference in this application, as if each document was individually incorporated by reference. In addition, it should be understood that after reading the above-mentioned teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (13)

  1. 一种功能性植物盐，其特征在于，所述功能性植物盐是如下制备的：将一种或多种植物提取物粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温下一次或分次烤制，获得功能性植物盐。A functional plant salt, characterized in that the functional plant salt is prepared as follows: one or more plant extract powders are mixed uniformly with fine salt at an addition amount of 1% to 40%, at 800- Roasted once or in multiples at a high temperature of 1400 ° C to obtain functional plant salts.
  2. 如权利要求1所述的功能性植物盐，其特征在于，所述植物提取物为竹提取物。The functional plant salt according to claim 1, wherein the plant extract is a bamboo extract.
  3. 如权利要求1所述的功能性植物盐，其特征在于，所述植物提取物选自下组：竹叶提取物、竹茹提取物、竹竿提取物、竹笋提取物、或其组合。The functional plant salt according to claim 1, wherein the plant extract is selected from the group consisting of bamboo leaf extract, bamboo rut extract, bamboo pole extract, bamboo shoot extract, or a combination thereof.
  4. 如权利要求1所述的功能性植物盐，其特征在于，所述植物提取物选自下组：竹叶黄酮、竹叶多酚、或其组合。The functional plant salt according to claim 1, wherein the plant extract is selected from the group consisting of bamboo leaf flavones, bamboo leaf polyphenols, or a combination thereof.
  5. 如权利要求1所述的功能性植物盐，其特征在于，所述细盐选自下组：海盐、湖盐、井盐、矿盐。The functional plant salt according to claim 1, wherein the fine salt is selected from the group consisting of sea salt, lake salt, well salt, and mineral salt.
  6. 如权利要求1所述的功能性植物盐，其特征在于，所述细盐为粗制海盐。The functional plant salt according to claim 1, wherein the fine salt is a crude sea salt.
  7. 如权利要求1所述的功能性植物盐，其特征在于，所述功能性植物盐为竹盐。The functional plant salt according to claim 1, wherein the functional plant salt is bamboo salt.
  8. 如权利要求1所述的功能性植物盐，其特征在于，所述植物提取物还包含梅提取物和/或松提取物。The functional plant salt according to claim 1, wherein the plant extract further comprises a plum extract and / or a pine extract.
  9. 一种权利要求1所述的功能性植物盐的制备方法，其特征在于，包括步骤：将一种或多种植物提取物粉末按照1％～40％的添加量与细盐混合均匀，于800-1400℃的高温下一次或分次烤制，获得功能性植物盐。A method for preparing a functional plant salt according to claim 1, comprising the step of: mixing one or more plant extract powders with fine salt at an addition amount of 1% to 40%, uniformly at Roasted once or in multiple times at a high temperature of -1400 ° C to obtain functional plant salts.
  10. 如权利要求9所述的方法，其特征在于，所述烤制在机械化烤炉中进行。The method according to claim 9, wherein said roasting is performed in a mechanized oven.
  11. 如权利要求9所述的方法，其特征在于，所述烤制在中频熔炼炉中进行。The method according to claim 9, wherein said roasting is performed in an intermediate frequency melting furnace.
  12. 一种食品或药品组合物，其特征在于，包含一种或多种权利要求1所述的功能性植物盐。A food or pharmaceutical composition, comprising one or more functional plant salts according to claim 1.
  13. 一种权利要求1所述的功能性植物盐的用途，其特征在于，用于制备选自下组的物质：药物助剂、保健食品/饮品、功能性食品/饮品、餐桌调味盐、烹饪盐、特殊用途化妆品或个人护理用品。The use of the functional plant salt according to claim 1, characterized in that it is used to prepare a substance selected from the group consisting of a pharmaceutical auxiliary, a health food / drink, a functional food / drink, a table seasoning salt, and a cooking salt , Special-purpose cosmetics or personal care products.
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