CN1082070C

CN1082070C - Natural pigment from aril of balsam pear

Info

Publication number: CN1082070C
Application number: CN98112414A
Authority: CN
Inventors: 谭新平
Original assignee: Individual
Current assignee: Individual
Priority date: 1998-03-27
Filing date: 1998-03-27
Publication date: 2002-04-03
Anticipated expiration: 2018-03-27
Also published as: CN1230574A

Abstract

The present invention utilizes the aril of balsam pear of cucurbitaceous plants as pigment raw material to extract prolycopene or natural pigment. The present invention is directly applied to food, feed, cosmetics and pharmaceuticals.

Description

The natural pigment of arillus in the Fructus Momordicae charantiae

The present invention relates to be the arillus of balsam pear (Momordica charatia L.) fruit of a kind of Hu Lu of utilization section (Cucurbitaceae) plant as the chromogen material, and the lycopene or the natural pigment that extract.

In recent years, the research of the physiologically active of natural pigment carotenoid, antitumous effect has become a hot topic.Maximum with the research β-Hu Luobusu at first, its anti-oxidant function and to the restraining effect of cancer existing many researchs are proved.Then, there are some researches show that xenthophylls etc. also has antitumous effect, particularly alpha-carotene that the better effect of β-Hu Luobusu is arranged on antitumous effect.Nearest studies show that lycopene has than β-Hu Luobusu and the more obvious effect of alpha-carotene, and becomes now the most a kind of pigment of noticeable carotenoid.But lycopene is because extraction cost height, output are few, thereby the research that has influenced it is carried out and used.Domestic teaching and scientific research unit all is to adopt pigment of synthetic colour the Sudan to replace in the standard of making lycopene during by line.Simultaneously, the carotenoid that comprises lycopene can not oneself synthesize generation in human body, the necessary carotenoids of its human body need absorb from food, though contain lycopene in many fruit things and the vegetables, but its content is generally few more than beta carotene, studies show that up to the present, there is following several plant content higher: tomato (3.1mg/100g, processing tomato (10.8mg/100g), watermelon (3.5-5.0mg/100g), the red trailing plants card (8-14mg/100g) of red series, pink natsudaidai (3.4mg/100g).But difficulty obtains high-grade lycopene from above-mentioned plant usually.Up to the present, lycopene does not almost have as gyp production.Therefore, people expect to find new containing the high plant of lycopene or turn out high-load kind.The balsam pear country of origin is considered to East India, subtropical zone, in the torrid zone in China, India, Asia and subtropical zone etc. extensively cultivated, it mainly is to utilize green fruit as vegetables, and the arillus in mellow fruit and the fruit does not utilize commercial obtaining.

Purpose of the present invention is exactly to utilize the arillus of Fructus Momordicae charantiae as the natural pigment raw material, produces lycopene or natural pigment with physiologically active and tight security.

Technical scheme of the present invention is, be pigment raw material with the arillus of the Fructus Momordicae charantiae of trigonella plant, and a kind of molecular formula that extracts is C ₄₀H ₅₆Molecular weight is 537, has two keys to add up to 13 in the molecular structure, and conjugated double bond is 11 lycopene or natural pigment.

Lycopene is extensively to be distributed in plant, a kind of in the carotenoid of animal kingdom is because of being at first separates from tomato (Lycopersicum, esculentum Mill), so be lycopene, also extensively be distributed in the tissue of human body, the crystallization of lycopene is garnet, examines under a microscope to be the crystallization of minute hand shape, 175 ℃ of fusing points, be soluble in chloroform, dithiocarbonic anhydride, benzene, dissolve in ether, sherwood oil, ethane, acetone etc., be insoluble in methyl alcohol, ethanol.

Therefore, the method that the present invention arillus from Fructus Momordicae charantiae is extracted above-mentioned pigment is: utilize organic solvent or supercritical co to extract, then the solution that extracts separated, make with extra care.

Alcohols such as organic solvent above-mentioned such as methyl alcohol, ethanol, propyl alcohol, and normal hexane, benzene, chloroform, acetone, ether, sherwood oil, vinyl acetic monomer etc. can use separately or be used.The solution that extracts concentrates earlier, uses suitable organic solvent diluting again behind finite concentration, re-uses chromatography and separates, makes with extra care.

Pigment provided by the invention directly applies in food, feed, makeup and the pharmaceuticals.

Below by embodiment the present invention is carried out more specific description, and protection scope of the present invention is not limited to this.

At first accompanying drawing is described:

Fig. 1,2 is the spectroscopic analysis figure that the photodiode array of the arillus pigment of Fructus Momordicae charantiae detects, wherein 1-xenthophylls; The 2-lycopene; 3, alpha-carotene; 4, β-Hu Luobusu;

Fig. 3,4 represents the component of carotenoid of arillus of differing maturity Fructus Momordicae charantiae and the variation of content by diagram and form respectively;

Fig. 5,6 represents the accelerate the ripening variation of carotenoid content of back arillus of prematurity Fructus Momordicae charantiae by diagram and form respectively.

Fig. 7 represents the intake and the experimental result that the prostate cancer relation takes place of lycopene.

Embodiment 1, the extraction of pigment and refining

1.1 the extraction of pigment:

Methyl alcohol with adding 40% in the arillus in the Fructus Momordicae charantiae adds a spot of antioxidant and magnesiumcarbonate simultaneously, grinds rapidly with tissue mincer or with mortar adding quartz sand.Then with the filtration of bleeding of acidproof filterability funnel, methanol wash with 40% several times after, falling this acidproof filterability funnel is contained on the filter flask of another cleaning, wash out pigment with the acetone extracting again, till filtrate is colourless, extract is poured in the separating funnel, and adding ether, the pigment commentaries on classics is dissolved in the ether, washs with saturated aqueous common salt and distilled water subsequently, and impel its layering.Then, give up the liquid of separating funnel lower floor, take out ether pigmentary layer liquid.Alkalize.(the arillus colour component of Fructus Momordicae charantiae is more simple, and alkalinization step can be omitted) alkalization is the methanol solution that adds 20% potassium hydroxide, is placed under the low temperature 16-18 hour, for reducing the oxidation of pigment, charges into nitrogen N in bottle ₂, the solution after the alkalization moves into separating funnel again, suitably adds ether, wash with saturated aqueous common salt, distilled water again, give up lower floor's liquid, take out upper strata ether pigmentary layer solution, after carrying out the dehydration more than 1 hour after the adding anhydrous sodium sulphate, filter, clean pigment on the anhydrous sodium sulphate with ether.Last constant volume.

1.2 the separation of pigment, refining

The ether pigment solution that falls constant volume is in nitrogen N ₂After concentrating down, use organic solvent (carrene: methyl alcohol is 1: 1) dissolving again after, the membrane filter by 0.45 μ m filters, and adopts the high performance liquid chromatography chromatography to separate at last, and is refining.

High performance liquid chromatography is to adopt the L-6200Intelligent pump of Hitachi, the L-4200UV-Detector of Hitachi and the D-2000Chromoto-Integrator of Hitachi; Photodiode array detector is Tianjin, island SPD-M6A, and post is ODS-2 anti-phase post (4 * 250mm GL Sciences Inc).The detection wavelength of high-performance liquid chromatograph is 451nm, and the detection wavelength of photodiode array detector is 195nm-600nm; Mobile phase A solution is the fine cyanogen methane of second: carrene: methyl alcohol=65: 25: 10, B solution are the fine cyanogen methane of 90% second.Adopt following condition to separate: 0-2 minute 50%A solution, 50%B solution; 2-40 minute A solution 100%, 40-50 minute, 50%A solution and 50%B solution.Flow velocity 0.8ml/min.

1.3 the analysis of pigment, evaluation

The analysis of each carotenoid, evaluation are that standard substance are separated with high performance liquid chromatography simultaneously with the sample pigment solution of extraction, compare its high-efficient liquid phase chromatogram and, adopt the computer system (software M6PACver2) of photodiode array detection to carry out the analysis of absorption spectrum respectively with after the high performance liquid chromatography separation.See Fig. 1, Fig. 2

Analytical results is as follows: the 1st peak among Fig. 1, Fig. 2 is xenthophylls, and second peak is lycopene; The 3rd peak is alpha-carotene; The 4th peak is β-Hu Luobusu.

1.4 pigment is quantitative

Lycopene quantitatively be after the lycopene standard substance that will purchase come accurately take by weighing, be diluted with an organic solvent to proper concn after, carry out efficient liquid phase chromatographic analysis.With concentration and the peak area that obtains and the peak area of sample lycopene of standard substance, obtain the concentration of sample.Other pigment concentration adopts the β-Hu Luobusu standard substance, obtains its concentration as stated above.

The degree of developing into of Fructus Momordicae charantiae (ripening degree) difference, the colour component of its arillus also has sizable different with content.For this reason, the inventor with oneself the cultivation Fructus Momordicae charantiae as research object, the fruit development stage has been mellow fruit from crude fruit has been divided into A to F6 stage, ripening degree A, B, C, D, E, F are respectively the fruit of blooming back 5 days, 9 days, 13 days, 18 days, 21 days, 22 days.The analytical results of the arillus of differing maturity fruit is as follows: few to the pigment content of ripening degree C arillus from ripening degree A, its major ingredient is an xenthophylls, secondly be alpha-carotene and β-Hu Luobusu, when fruit development arrives ripening degree D, its lycopene begins to occur, when the ripening degree E, the rapid straight line of lycopene content rises, and reaches high-content 64.75mg/100g.Have the xenthophylls of minute quantity to be detected simultaneously, when fruit became overdone ripening degree F, lycopene content descended on the contrary to some extent, sees shown in Fig. 3,4.

By above analysis as can be known, the most suitable as pigment source with the fruit of ripening degree E.

Embodiment two: extract pigment after utilizing green fruit to accelerate the ripening

Be put into 20 ℃, 30 ℃ storages respectively and accelerated the ripening growing to a certain degree balsam pear (ripe degree C).The fruit branch is clipped to 8 days and fruit flavescence in 4 days, and the arillus of fruit also reddens.The result shows by analysis, and fruit is placed in 30 ℃, and the effect of accelerating the ripening is best, and pigment content is the highest, shown in Fig. 5,6.But the fruit than natural maturity on the Stem of Balsampear lacks about 1/3rd.Carry out taking out, carry, make with extra care, analyzing of pigment according to embodiment 1 same method then.

Can be used as the pigment additive by the lycopene that embodiment one or two extracted and be applied to fields such as food, feed, makeup, pharmaceuticals.Lycopene uses as tinting material, and its tone can change to reddish orange from yellow with change in concentration, takes on a red color in the dispersed system of ultra-fine grain, uses and can adjust as required.

The look of tomato processed goods is to weigh one of most important index of estimating the goods quality.And lycopene has sanatory physiological function and cancer resistance on health except regulating color.Become the most noticeable pigment.Be that example describes below with the pasta sauce.

The pigment adjustment (enhancing) of tomato ketchup: (trade name: the anticancer tomato ketchup) interpolation (being adjusted to 15-20mg/100g) of 3.6 liters of cassia 28g lycopenes of tomato puree (proportion 1.022) 90kg basyleave 14g granulated sugar 5.4kg celery seed 6g salt 0.9kg nutmeg 3g onion 1.8kg cloves sub-6g pimiento 19g allspice 6g white pepper 15g vinegar (containing 5% acetic acid)

The natural pigment of arillus is that the arillus of employing natural phant Fructus Momordicae charantiae is a pigment raw material in the Fructus Momordicae charantiae provided by the invention, its lycopene content height, higher 20 times than general edible ripe tomato (3.1mg/100g), be to find that so far content is the highest in the plant tissue, and the component of pigment is also very simple, other pigment content is few, therefore, be easy to the production of lycopene and refining, its production cost also can be improved significantly, helps the enforcement of industrial production.Lycopene uses as tinting material, and its tone can change to reddish orange from yellow along with change in concentration, takes on a red color in the dispersed system of ultra-fine grain, use and can adjust as required, and on yellow tone, its effect is about 8 times an of β-Hu Luobusu.

The security of lycopene in the mouse pharmacological testing, LD50 (Lethal Dose 50) more than 3g/kg, to big white mouse, the long-term feeding trial of dog, also prove do not have fully toxic.

Lycopene has anti-oxidant function, is more than 2 times of β-Hu Luobusu, and more than 100 times of vitamin-E are the most resultful a kind of in carotenoid.

Lycopene has very strong antitumous effect, to studies have shown that of lycopene, the rejection ratio alpha-carotene and the β-Hu Luobusu of cancer cell multiplication is eager to excel recently.

1986-1992 6 in the period of, carried out the perspective epidemiology survey of " carotenoid and vitamin A are taken in the relation with prostate cancer ".The male sex's 51529 examples in age 40-75 year have been investigated altogether.The result shows that the absorption of having only lycopene and the risk level that prostate cancer takes place have the negative correlation of significance, along with the intake increase of lycopene every day, the risk level of generation prostate cancer reduce.The experimental result of its lycopene sees for details shown in Figure 7.

Therefore natural lycopene provided by the invention is not only safe, tone range is wide, and be to have anti-oxidant function, the pigment of physiological properties such as antitumous effect will have application more and more widely at aspects such as heath food, feed, makeup, pharmaceuticals.

Claims

1, a kind of method of from the arillus of Fructus Momordicae charantiae, extracting lycopene, utilize organic solvent or supercritical co to extract, then the soln using chromatography that extracts is separated, refining, it is characterized in that: the extraction process of pigment is, with the methyl alcohol that adds 40% in the arillus in the Fructus Momordicae charantiae, add a spot of antioxidant and magnesiumcarbonate simultaneously, grind rapidly with tissue mincer or with mortar adding quartz sand, then with the filtration of bleeding of acidproof filterability funnel, methanol wash with 40% several times after, this acidproof filterability funnel is contained on the filter flask of another cleaning, wash out pigment with the acetone extracting again, till filtrate is colourless, extract is poured in the separating funnel, and adding ether, the pigment commentaries on classics is dissolved in the ether, wash with saturated aqueous common salt and distilled water subsequently, and impel its layering, the liquid of giving up separating funnel lower floor then takes out ether pigmentary layer liquid, alkalizes, alkalization is the methanol solution of the potassium hydroxide of adding 20%, be placed under the low temperature 16-18 hour,, in bottle, charge into nitrogen N for reducing the oxidation of pigment ₂The solution of alkalization moves into separating funnel again, suitably add ether, wash with saturated aqueous common salt, distilled water again, give up lower floor's liquid, take out upper strata ether pigmentary layer solution, after carrying out the dehydration more than 1 hour after the adding anhydrous sodium sulphate, filter, clean pigment on the anhydrous sodium sulphate, last constant volume with ether; Separation, the treating process of pigment be, with the ether pigment solution of constant volume in nitrogen N ₂After concentrating down, again with carrene: behind the organic solvent dissolution that methyl alcohol is made into,, adopt the high performance liquid chromatography chromatography to separate, make with extra care at last by the membrane filter filtration of 0.45um in 1: 1 ratio.

According to the method for claim 1, it is characterized in that 2, the detection wavelength of the high-performance liquid chromatograph that the high performance liquid chromatography chromatography is adopted is 451nm, the detection wavelength of the photodiode array detector of employing is 195nm-600nm; Mobile phase A solution is second eyeball cyanogen methane: carrene: methyl alcohol=65: 25: 10, B solution are 90% second eyeball cyanogen methane, and adopt following condition to separate: 0-2 minute 50%A solution, 50%B solution; 2-4.0 minute A solution 100%, 40-50 minute, 50%A solution and 50%B solution; Flow velocity 0.8ml/min.

3, according to the method for claim 1, it is characterized in that, utilize immature Fructus Momordicae charantiae to extract pigment, be to be put into 20 ℃, 30 ℃ storages respectively and to be accelerated the ripening with growing to a certain degree balsam pear, the fruit branch is clipped to 8 days and flavescence in 4 days, the arillus of fruit also reddens, and then extracts, separates, makes with extra care.