CN107976499A - A kind of method of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization - Google Patents
A kind of method of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization Download PDFInfo
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Abstract
The method of gamma aminobutyric acid content, is related to gamma aminobutyric acid content detection technical field in a kind of efficiently binary liquid chromatography detection livestock and poultry organization, and this method is to weigh livestock and poultry organization, adds liquid nitrogen, and grinding is so repeated multiple times, until livestock and poultry organization crushes completely；Sample diluting liquid is added, high speed centrifugation, takes supernatant to cross miillpore filter；Filtrate is freeze-dried, adds and adjusts solution, it is cold dry；Derivatization reagent is added, is done after room temperature derivative through cold, obtains sample；Sample is redissolved with sample diluting liquid, obtains sample, sample and standard specimen injection high performance liquid chromatograph are analyzed, obtain the chromatogram of sample and standard specimen, gamma aminobutyric acid content in sample is calculated according to chromatogram.The detection method is made derivating agent using phenyl isothiocyanate and is reacted with the amino acid in livestock and poultry organization, and derivative is detected after the separation of binary efficient liquid phase with UV detector, can directly detect the gamma aminobutyric acid residual in livestock and poultry organization, and testing result accuracy is high.
Description
Technical field
The present invention relates to alpha-aminobutyric acid content detection technique field, and more particularly to a kind of efficiently binary liquid chromatogram
The method of alpha-aminobutyric acid content in method detection livestock and poultry organization.
Background technology
γ-aminobutyric acid (γ-Aminobutyric acid, GABA) is also known as amino acid injection-800, nipecotic acid, its molecular formula is
C4H9NO2, molecular weight 130.12.γ-aminobutyric acid is a kind of nonprotein amino acid, is distributed widely in animal and plant body.Mesh
Before, it is a kind of important suppressive neurotransmitter that γ-aminobutyric acid, which medically studies more deep effect, has and promotes brain
Vigor, calm the nerves, adjust hormone secretion, improve the biological functions such as lipid-metabolism, blood pressure lowering.Based on the above-mentioned of γ-aminobutyric acid
Function, it is widely used in livestock and poultry breeding industry, γ-aminobutyric acid primarily as a kind of maincenter sedative, for example, γ-
Aminobutyric acid is added in pannage, for regulating and controlling to the growth performance of pig.
Research GABA in livestock and poultry breeding industry in application, it is generally necessary to being detected to the GABA in animal products.
Currently used GABA detection methods have thin-layer chromatography, paper chromatography, colorimetric method, amino-acid analyzer measure, Capillary Electrophoresis-
Electrochemical Detection, high performance liquid chromatography (High Performance Liquid Chromatography, HPLC).Various sides
Method has its advantage and disadvantage, wherein thin-layer chromatography method high sensitivity, easy to detect, but detects time-consuming long；Paper electrophoresis method
Cost is relatively low, and without expensive reagent and instrument, shortcoming is the shadow for needing to control temperature, pH and filter paper nature etc.
Ring, operating process is cumbersome；Paper chromatography is easy and effective low-cost, and shortcoming is that precision is not high, and detection time is longer；Amino acid
Analyzer detection is accurate, simplicity is time saving, stability is good, and shortcoming is that equipment is expensive, the purity requirement to sample is very high, determination condition
Need strictly to control；Capillary electrophoresis high sensitivity, cost are low, reagent dosage is few, but operating process is excessively complicated；Efficiently
Liquid chromatography accuracy and sensitivity are very high, can meet the needs detected during most of research and production.
Both at home and abroad the detection object of the high performance liquid chromatography of measure γ-aminobutyric acid be mostly alpha-aminobutyric acid content compared with
High brain tissue or serum, and multi-purpose o-phthalaldehyde is sent out as derivating agent under catalyst action with the amino acid in tissue
Raw derivative reaction, the derivative of generation are separated through efficient liquid phase, then are detected by Electrochemical Detection or fluorescence detector.It is this
The testing result accuracy of detection method is relatively low, and can not directly obtain livestock and poultry major product --- γ-amino fourth in meat
Acid content.
The content of the invention
It is an object of the invention to provide γ-aminobutyric acid in a kind of efficiently binary liquid chromatography detection livestock and poultry organization to contain
The method of amount, can directly detect the γ-aminobutyric acid residual in livestock and poultry organization, and testing result accuracy is high.
The present invention is solved its technical problem and is realized using following technical scheme.
The method that the present invention proposes alpha-aminobutyric acid content in a kind of efficiently binary liquid chromatography detection livestock and poultry organization,
It comprises the following steps：
Livestock and poultry organization is weighed, adds liquid nitrogen, is ground after livestock and poultry organization cooling is hard, adds liquid nitrogen, grinding, such as
This is repeated multiple times, until livestock and poultry organization crushes completely；Sample diluting liquid is added, high speed centrifugation, takes supernatant to cross miillpore filter, obtains
Filtrate；
Filtrate is freeze-dried, adds and adjusts solution, freeze-drying, it is by methanol, sodium acetate and triethylamine to adjust solution
In mass ratio 2~3：2~3：1 composition；Derivatization reagent is added, derivatization reagent is by phenyl isothiocyanate, methanol, second
Alcohol, triethylamine and water are according to mass ratio 1~2：5~6：1~2：1~2：1 composition, through freeze-drying after room temperature derivative, obtains sample
Product；
Sample is redissolved with sample diluting liquid, obtains sample, sample and standard specimen injection high performance liquid chromatograph are analyzed,
The chromatogram of sample and standard specimen is obtained, γ-Gamma-propalanine content in sample is calculated according to chromatogram.
Further, in present pre-ferred embodiments, sample diluting liquid is the Na of 4~6mmol/l2HPO4Buffer molten
Liquid, pH=7~8.
Further, in present pre-ferred embodiments, the dosage of livestock and poultry organization and the sample diluting liquid added for the first time
Than for 0.1g：350~450 μ L.
Further, in present pre-ferred embodiments, filtrate, adjust solution, derivatization reagent and the sample of redissolution
The amount ratio of dilution is 2~3：1：2~3：3~5.
Further, in present pre-ferred embodiments, ultracentrifugal method is：In 10000~13000r/min from
The heart 3~10 minutes.
Further, in present pre-ferred embodiments, the filter opening of miillpore filter is 0.4~0.5 μm of filter membrane.
Further, in present pre-ferred embodiments, before detection, the livestock and poultry organization removed by livestock and poultry need to be put into liquid nitrogen
In, it is then transferred to freezen protective in less than -20 DEG C of refrigerator-freezer.
Further, in present pre-ferred embodiments, sample is placed in 2~6 DEG C and preserves before redissolution.
Further, in present pre-ferred embodiments, the chromatographic condition of high performance liquid chromatograph is：Chromatographic column is amino
Sour dedicated columns；Mobile phase by mobile phase A and/or Mobile phase B form carry out gradient elution, mobile phase A be by acetonitrile, 65~
75mmol/L acetate buffer solutions and ethylenediamine tetra-acetic acid in mass ratio 20~30：900~1050：0.25 composition, Mobile phase B be by
Acetonitrile, water and methanol in mass ratio 40~50：40：10~20 compositions；Flow velocity is 1~1.5ml/min；Ultraviolet detection wavelength is
254nm；Column temperature is 40~50 DEG C；Sample size is 10~20 μ L.
Further, in present pre-ferred embodiments, according to sample and the appearance time of standard specimen, identify in sample whether
Contain γ-aminobutyric acid；The peak area in chromatogram by contrasting sample and standard specimen, calculates γ-aminobutyric acid in sample and contains
Amount.
The method of alpha-aminobutyric acid content in the efficient binary liquid chromatography detection livestock and poultry organization of the embodiment of the present invention
Beneficial effect is：The side of alpha-aminobutyric acid content in the efficient binary liquid chromatography detection livestock and poultry organization of the embodiment of the present invention
Method is to weigh livestock and poultry organization, adds liquid nitrogen, is ground after livestock and poultry organization cooling is hard, adds liquid nitrogen, grinding, so anti-
It is multiple multiple, until livestock and poultry organization crushes completely；Sample diluting liquid is added, high speed centrifugation, takes supernatant to cross miillpore filter, must filter
Liquid；Filtrate is freeze-dried, adds and adjusts solution, freeze-drying；Derivatization reagent is added, it is dry through freezing after room temperature derivative
It is dry, obtain sample；Sample is redissolved with sample diluting liquid, obtains sample, sample and standard specimen injection high performance liquid chromatograph are divided
Analysis, obtains the chromatogram of sample and standard specimen, and alpha-aminobutyric acid content in sample is calculated according to chromatogram.The detection method uses different
Thiocyanic acid phenyl ester is made derivating agent and is reacted with the amino acid in livestock and poultry organization, and derivative is after the separation of binary efficient liquid phase with ultraviolet inspection
Device detection is surveyed, can directly detect the γ-aminobutyric acid residual in livestock and poultry organization, testing result accuracy is high.
Brief description of the drawings
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it will be appreciated that the following drawings illustrate only certain embodiments of the present invention, therefore be not construed as pair
The restriction of scope, for those of ordinary skill in the art, without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the chromatogram of pork sample in the embodiment of the present invention 1；
Fig. 2 is the chromatogram of γ-aminobutyric acid standard specimen in the embodiment of the present invention 1.
Embodiment
, below will be in the embodiment of the present invention to make the purpose, technical scheme and advantage of the embodiment of the present invention clearer
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, builds according to normal condition or manufacturer
The condition of view carries out.Reagents or instruments used without specified manufacturer, is the conventional production that can be obtained by commercially available purchase
Product.
Below to γ-Gamma-propalanine content in the efficient binary liquid chromatography detection livestock and poultry organization of the embodiment of the present invention
Method be specifically described.
The embodiment of the present invention provides γ-Gamma-propalanine content in a kind of efficiently binary liquid chromatography detection livestock and poultry organization
Method, it comprises the following steps：
(1) tissue is handled：The livestock and poultry organization removed by livestock and poultry need to be put into liquid nitrogen, be then transferred to less than -20 DEG C of refrigerator-freezer
Middle freezen protective, livestock and poultry organization refer to pig rib meat, abdominal fat, liver or brain.Above-mentioned livestock and poultry organization is weighed, liquid nitrogen is added, treats
It is ground after livestock and poultry organization cooling is hard, adds liquid nitrogen, grinding, it is so repeated multiple times, until livestock and poultry organization crushes completely；
Sample diluting liquid is added, sample diluting liquid can be the Na of 4~6mmol/l2HPO4Buffer solution, pH=7~8, poultry
Fowl is organized and the amount ratio of sample diluting liquid is 0.1g：350~450 μ L, high speed centrifugation, specifically in 10000~13000r/
Min is centrifuged 3~10 minutes, is taken supernatant to cross 0.4~0.5 μm of miillpore filter, is obtained filtrate.
(2) it is derivative：Filtrate is freeze-dried, adds and adjusts solution, be freeze-dried again, appearance may be sticked to for neutralizing
Remaining acid on device, it is by methanol, sodium acetate and triethylamine in mass ratio 2~3 to adjust solution：2~3：1 composition；Add and spread out
Biochemical reagents, derivatization reagent are according to mass ratio 1~2 by phenyl isothiocyanate, methanol, ethanol, triethylamine and water：5~6：
1~2：1~2：1 composition, the amount ratio of filtrate, adjusting solution and derivatization reagent is 2~3： 1：2~3, derive 15 in room temperature
Through freeze-drying after~30min, sample is obtained, is placed in 2~6 DEG C of refrigerator and preserves.
(3) machine is analyzed on：Sample is redissolved with sample diluting liquid, adjusts the dosage of solution and the sample diluting liquid of redissolution
Than for 1：3~5, obtain sample, by sample and standard specimen injection SSI PC-2001 high performance liquid chromatographs analyzed, obtain sample and
The chromatogram of standard specimen, calculates alpha-aminobutyric acid content in sample, specific method is according to the chromatogram of sample and standard specimen：According to
The appearance time of sample and standard specimen, identifies in sample whether contain γ-aminobutyric acid, then by contrasting the chromatography of sample and standard specimen
Peak area in figure, calculates alpha-aminobutyric acid content in sample.
In the present embodiment, the chromatographic condition of high performance liquid chromatograph is：Chromatographic column is amino acid dedicated columns；Mobile phase be by
Mobile phase A and/or Mobile phase B composition carry out gradient elution, and within the time of 0~30min, mobile phase A accounting is reduced by 100%
For 0, the accounting of Mobile phase B is 100% by 0 increase and decrease, and mobile phase A is by acetonitrile, 65~75mmol/L acetate buffer solutions (PH=
And ethylenediamine tetra-acetic acid in mass ratio 20~30 6.5)：900~1050：0.25 composition, Mobile phase B is by acetonitrile, water and methanol
In mass ratio 40~50：40：10~20 compositions；Flow velocity is 1~1.5ml/min；Ultraviolet detection wavelength is 254nm；Column temperature is 40
~50 DEG C；Sample size is 10~20 μ L.
High performance liquid chromatography is on the basis of classical liquid chromatography, and gas-chromatography reason is introduced in the later stage sixties
By and develop rapidly.The difference of it and classical liquid chromatography is that filler particles are small and uniform, and little particle has Gao Zhu
Effect, but can cause high-drag, need to use high-pressure delivery mobile phase, therefore also known as high pressure lipuid chromatography (HPLC) (High Pressure
Liquid Chromatography, HPLC), but because analyze speed is fast and referred to as high-speed liquid chromatography (High Speed
Liquid Chromatography, HSLP), it is also referred to as modern liquid chromatography.
The characteristics of high performance liquid chromatography and advantage：
High pressure：Pressure is up to 150~300Kg/cm2.Every meter of decompression of chromatographic column is more than 75Kg/cm2.
At a high speed：Flow velocity is 0.1~10.0ml/min.
Efficiently：Up to 5000 every meter of column plates, composition is separated up to 100 kinds at the same time in a column.
High sensitivity：UV detector sensitivity is up to 0.01ng, while it is few to consume sample.
High performance liquid chromatography has the advantage that compared with classical liquid chromatogram：
Speed is fast：Usually one sample of analysis can even be completed in 15~30min, some samples in 5min.
High resolution：Stationary phase and mobile phase may be selected to reach optimal separation effect.
High sensitivity：UV detector is up to 0.01ng, and fluorescence and electrochemical detector are up to 0.1pg.
Pillar can Reusability：Different compounds is separated with a root chromatogram column.
Sample size is few, easily recycling：Sample is not destroyed after chromatographic column, can be collected one-component or be prepared.
Embodiment 1
Researcher is when the production performance of pork pig in the case of studying γ-aminobutyric acid to heat stress influences, according to this reality
Apply the residual quantity of γ-aminobutyric acid in the detection method detection Pork Tissue of example.
First, instrument and reagent
1st, instrument：Highly effective liquid phase chromatographic system, including two pumps, UV detector, automatic gradient controller, temperature control
Instrument and column oven；Sampling valve；Chromatographic work station, amino acid detection column.
2nd, γ-aminobutyric acid standard items, phenyl isothiocyanate (derivating agent)；Acetonitrile, methanol (chromatographically pure), sodium acetate (point
Analyse pure), level-one water.
2nd, chromatographic condition
Chromatographic column：Amino acid dedicated columns；
Mobile phase：Mobile phase A is acetonitrile：70mmol/L acetate buffer solutions (PH=6.5)：Ethylenediamine tetra-acetic acid=25：
975：0.25 (mass ratio)；
Mobile phase B is acetonitrile：Water：Methanol=45：40：15 (mass ratioes)；
Flow velocity is 1ml/min；
Ultraviolet detection wavelength 254nm；
45 DEG C of column temperature；
20 μ L of sample size；
Condition of gradient elution such as following table：
1 gradient elution program table of table
| Time min | A% | B% |
| 0 | 100 | 0 |
| 13.5 | 97 | 3 |
| 20 | 96 | 4 |
| 20.5 | 20 | 80 |
| 22.5 | 0 | 100 |
| 23 | 100 | 0 |
| 30 | 100 | 0 |
3rd, solution is prepared
Sample diluting liquid：The Na of 5mmol/l2HPO4Buffer solution, pH=7.4；
Derivatization reagent：Phenyl isothiocyanate：Methanol：Ethanol：Triethylamine：Water=1：6：1：1：1 (mass ratio), needs fresh
Prepare；
Adjust solution：Methanol：Sodium acetate：Triethylamine=2：2：1 (mass ratio).
4th, specific experiment method：
(1) sample treatment
Take pork to be put into liquid nitrogen rapidly, be then transferred to freezen protective in -20 DEG C of refrigerator-freezers, wait to be detected.
The Pork Tissue for weighing 0.1g is placed in small mortar, pours into liquid nitrogen, is ground, is crushed after tissue cooling is hard
After add liquid nitrogen grinding, so repeatedly 3 times, Pork Tissue is crushed completely.
The tissue of crushing is completely transferred in centrifuge tube by the sample diluting liquid for adding 400 μ L, and 5 are centrifuged in 11000r/min
Minute, take supernatant to cross 0.45 μm of filter membrane, obtain filtrate.
(2), it is derivative
Take in 20 μ L implantation glass test tubes of filtrate, be freeze-dried, then add and adjust 10 μ L of solution, freeze again, use
Remaining acid on test tube may be sticked in neutralizing；20 μ L derivatization reagents are added, passes through and freezes after room temperature derivative 20min, obtain sample
Product, are placed in 4 DEG C of refrigerators and preserve.
(3), upper machine analysis
With 50 μ L samples dilutions redissolve, obtain sample, with micro syringe inject 20 μ L sample and 20 μ L standard specimen in
Analyzed on SSI PC-2001 high performance liquid chromatographs, obtain the chromatogram of sample and standard specimen, the chromatogram of pork sample is as schemed
Shown in 1, as shown in Fig. 2, according to sample and the appearance time of standard specimen, identify in sample is the chromatogram of γ-aminobutyric acid standard specimen
It is no containing γ-aminobutyric acid, then the peak area in chromatogram by contrasting sample and standard specimen calculates gamma-amino fourth in sample
Acid content.
In conclusion γ-aminobutyric acid contains in the efficient binary liquid chromatography detection livestock and poultry organization of the embodiment of the present invention
The method of amount can directly detect the γ-aminobutyric acid residual in livestock and poultry organization, and testing result accuracy is high.
Embodiments described above is part of the embodiment of the present invention, instead of all the embodiments.The reality of the present invention
The detailed description for applying example is not intended to limit the scope of claimed invention, but is merely representative of the selected implementation of the present invention
Example.Based on the embodiments of the present invention, those of ordinary skill in the art are obtained without creative efforts
Every other embodiment, belongs to the scope of protection of the invention.
Claims (10)
1. a kind of method of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization, it is characterised in that its
Comprise the following steps：
Livestock and poultry organization is weighed, adds liquid nitrogen, is ground after livestock and poultry organization cooling is hard, adds liquid nitrogen, grinding, such as
This is repeated multiple times, until the livestock and poultry organization crushes completely；Sample diluting liquid is added, high speed centrifugation, takes supernatant to cross micropore filter
Film, obtains filtrate；
The filtrate is freeze-dried, adds and adjusts solution, freeze-drying, the adjusting solution is by methanol, sodium acetate and three
Ethamine in mass ratio 2~3：2~3：1 composition；Add derivatization reagent, the derivatization reagent be by phenyl isothiocyanate,
Methanol, ethanol, triethylamine and water are according to mass ratio 1~2：5~6：1~2：1~2：1 composition, it is dry through freezing after room temperature derivative
It is dry, obtain sample；
Sample is redissolved with sample diluting liquid, obtains sample, sample and standard specimen injection high performance liquid chromatograph are analyzed, obtain sample
The chromatogram of product and standard specimen, alpha-aminobutyric acid content in sample is calculated according to the chromatogram.
2. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that the sample diluting liquid is the Na of 4~6mmol/l2HPO4Buffer solution, pH=7~8.
3. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that the amount ratio of the livestock and poultry organization and the sample diluting liquid added for the first time is 0.1g：350~450
μL。
4. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that the filtrate, the dosage for adjusting solution, the derivatization reagent and the sample diluting liquid of redissolution
Than for 2~3：1：2~3：3~5.
5. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that ultracentrifugal method is：Centrifuged 3~10 minutes in 10000~13000r/min.
6. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that the filter opening of the miillpore filter is 0.4~0.5 μm of filter membrane.
7. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that before detection, the livestock and poultry organization removed by livestock and poultry need to be put into liquid nitrogen, be then transferred to less than -20 DEG C of ice
Freezen protective in cabinet.
8. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that the sample is placed in 2~6 DEG C and preserves before redissolution.
9. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that the chromatographic condition of the high performance liquid chromatograph is：Chromatographic column is amino acid dedicated columns；Mobile phase be by
Mobile phase A and/or Mobile phase B composition carry out gradient elution, mobile phase A be by acetonitrile, 65~75mmol/L acetate buffer solutions and
Ethylenediamine tetra-acetic acid in mass ratio 20~30：900~1050：0.25 composition, Mobile phase B is by quality by acetonitrile, water and methanol
Than 40~50：40：10~20 compositions；Flow velocity is 1~1.5ml/min；Ultraviolet detection wavelength is 254nm；Column temperature is 40~50 DEG C；
Sample size is 10~20 μ L.
10. the side of alpha-aminobutyric acid content in efficiently binary liquid chromatography detection livestock and poultry organization according to claim 1
Method, it is characterised in that according to sample and the appearance time of standard specimen, identify in sample whether contain γ-aminobutyric acid；Pass through contrast
Peak area in the chromatogram of sample and standard specimen, calculates alpha-aminobutyric acid content in sample.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108693278A (en) * | 2018-07-06 | 2018-10-23 | 漳州傲农牧业科技有限公司 | The assay method of IgG content in a kind of Swine plasma protein |
| CN111044640A (en) * | 2019-12-31 | 2020-04-21 | 杭州康德权饲料有限公司 | Method for determining content of gamma-aminobutyric acid in feed additive by GC (gas chromatography) method |
| CN114113349A (en) * | 2020-08-26 | 2022-03-01 | 湖南农业大学 | Method for simultaneously determining 5 amino acid neurotransmitters in biological matrix based on two-dimensional liquid chromatography-ultraviolet derivatization method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102539600A (en) * | 2011-12-23 | 2012-07-04 | 上海中医药大学附属岳阳中西医结合医院 | Method for detecting aminobutyric acid in sample by high efficiency liquid chromatography |
| CN106645516A (en) * | 2016-12-29 | 2017-05-10 | 香港科技大学 | Method for detecting neurotransmitter in hair |
| CN107064340A (en) * | 2017-03-13 | 2017-08-18 | 瀚盟测试科技（天津）有限公司 | A kind of method of GABA content in detection tealeaves |
-
2017
- 2017-11-27 CN CN201711205670.7A patent/CN107976499A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102539600A (en) * | 2011-12-23 | 2012-07-04 | 上海中医药大学附属岳阳中西医结合医院 | Method for detecting aminobutyric acid in sample by high efficiency liquid chromatography |
| CN106645516A (en) * | 2016-12-29 | 2017-05-10 | 香港科技大学 | Method for detecting neurotransmitter in hair |
| CN107064340A (en) * | 2017-03-13 | 2017-08-18 | 瀚盟测试科技（天津）有限公司 | A kind of method of GABA content in detection tealeaves |
Non-Patent Citations (3)
| Title |
|---|
| MARÍA-CONCEPCIÓN ARISTOY 等: "Deproteinization Techniques for HPLC Amino Acid Analysis in Fresh Pork Muscle and Dry-Cured Ham", 《JOURNAL OR AGRICULTURAL AND FOOD CHEMISTRY》 * |
| 谭力 等: "人胃粘膜中γ-氨基丁酸和谷氨酸含量的高效液相色谱法测定", 《色谱》 * |
| 钱爱萍 等: "家禽肉中氨基酸组成及营养评价", 《中国农学通报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108693278A (en) * | 2018-07-06 | 2018-10-23 | 漳州傲农牧业科技有限公司 | The assay method of IgG content in a kind of Swine plasma protein |
| CN111044640A (en) * | 2019-12-31 | 2020-04-21 | 杭州康德权饲料有限公司 | Method for determining content of gamma-aminobutyric acid in feed additive by GC (gas chromatography) method |
| CN114113349A (en) * | 2020-08-26 | 2022-03-01 | 湖南农业大学 | Method for simultaneously determining 5 amino acid neurotransmitters in biological matrix based on two-dimensional liquid chromatography-ultraviolet derivatization method |
| CN114113349B (en) * | 2020-08-26 | 2024-01-23 | 湖南农业大学 | Method for simultaneously measuring 5 amino acid neurotransmitters in biological matrix based on two-dimensional liquid chromatography-ultraviolet derivatization method |
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