CN113341029B - Method for detecting content of gamma-aminobutyric acid in cosmetics - Google Patents
Method for detecting content of gamma-aminobutyric acid in cosmetics Download PDFInfo
- Publication number
- CN113341029B CN113341029B CN202110780174.4A CN202110780174A CN113341029B CN 113341029 B CN113341029 B CN 113341029B CN 202110780174 A CN202110780174 A CN 202110780174A CN 113341029 B CN113341029 B CN 113341029B
- Authority
- CN
- China
- Prior art keywords
- solution
- column
- gamma
- acid
- buffer solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/067—Preparation by reaction, e.g. derivatising the sample
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a detection method of gamma-aminobutyric acid in cosmetics, which comprises the following steps: preparation of a control: dissolving gamma-aminobutyric acid reference substance by using hydrochloric acid, and filtering to obtain gamma-aminobutyric acid reference substance solution; preparation of test article: dissolving a cosmetic sample with hydrochloric acid, and filtering to obtain a sample solution; preparation of a blank solution: filtering hydrochloric acid to obtain a blank solution; and (3) measuring: and detecting the reference substance solution, the test substance solution and the blank solution by using a separation column and a post-column derivatization reaction column, thereby obtaining the content of gamma-aminobutyric acid in the cosmetic test substance. The detection method is simple to operate, sensitive in response, high in accuracy and good in repeatability, and can be used for detecting the content of gamma-aminobutyric acid in cosmetics.
Description
Technical Field
The invention relates to the technical field of gamma-aminobutyric acid detection, in particular to a method for detecting the content of gamma-aminobutyric acid in cosmetics.
Background
Gamma-aminobutyric acid (GABA) also known as aminobutyric acid, 4-aminobutyric acid, piperidine acid, molecular formula C 4 H 9 NO 2 Molecular weight 103.1. Gamma-aminobutyric acid is an inhibitory neurotransmitter of the mammalian central nervous system, widely distributed in prokaryotes and eukaryotes, and has been studied to demonstrate its efficacy in relieving anxiety, lowering blood pressure, improving sleep, removing wrinkles and whitening. By utilizing the property of the gamma-aminobutyric acid at home and abroad, a series of products are developed to hope for lovers, but when the concentration of the gamma-aminobutyric acid reaches the wrinkle removing concentration or is expected to reach the better wrinkle removing concentration, discomfort symptoms such as tingling, itching, burning and the like can be generated when the gamma-aminobutyric acid contacts with skin, and the side effect has positive correlation with the concentration of the gamma-aminobutyric acid. In order to make the gamma-aminobutyric acid have wider application in the skin care field, the proper dosage of the gamma-aminobutyric acid needs to be ensured.
Because gamma-aminobutyric acid has no ultraviolet absorption, the content of gamma-aminobutyric acid needs to be detected by using a high performance liquid chromatography and reacts with a derivatization reagent to generate a compound with ultraviolet absorption, but the pre-column derivatization reaction is easy to be interfered by the outside to form multi-stage derivatization, the repeatability of a detection result is poor and the accuracy is low after separation by a chromatographic column, and the detection of the content of gamma-aminobutyric acid is influenced. The post-column derivatization cation exchange chromatography is characterized in that gamma-aminobutyric acid is separated by a cation exchange chromatography column and then is subjected to derivatization treatment, and has the advantages of high reaction speed, good stability of a derivatization product and the like. When the mobile phase commonly used for detecting and separating amino acid by an amino acid analyzer and an elution program are used for detecting gamma-aminobutyric acid, the peak time of gamma-aminobutyric acid is about 1.5min, the gamma-aminobutyric acid is extremely easily influenced by a solvent peak, the peak area is small, and the detection accuracy of gamma-aminobutyric acid is easily influenced by baseline fluctuation.
Disclosure of Invention
As described above, in order to solve the above technical problems, the present invention provides a method for detecting gamma-aminobutyric acid in cosmetics, which can greatly improve the sensitivity of detection while ensuring the accuracy of detection results, and has the advantages of simple sample processing and convenient operation.
The specific technical scheme of the invention is as follows:
1. a method for detecting gamma-aminobutyric acid in cosmetics, which comprises the following steps:
preparation of a control: dissolving gamma-aminobutyric acid reference substance by using hydrochloric acid, and filtering to obtain gamma-aminobutyric acid reference substance solution;
preparation of test article: dissolving a cosmetic sample with hydrochloric acid, and filtering to obtain a sample solution;
preparation of a blank solution: filtering hydrochloric acid to obtain a blank solution;
and (3) measuring: and detecting the reference substance solution, the test substance solution and the blank solution by using a separation column and a post-column derivatization reaction column, thereby obtaining the content of gamma-aminobutyric acid in the cosmetic test substance.
2. The detection method according to item 1, wherein the concentration of the hydrochloric acid is 0.02mol/L to 0.06mol/L.
3. The detection method according to item 1 or 2, wherein the dissolution is a water bath dissolution, preferably a dissolution time of 4 to 6 hours, further preferably a dissolution temperature of 60 to 80 ℃.
4. The detection method according to any one of claims 1 to 3, wherein the cosmetic is a cream-milky or liquid-like cosmetic, preferably, the cream-milky cosmetic is an emulsion, an eye cream, or a facial cleanser; preferably, the liquid cosmetic is essential oil, toner or bath lotion.
5. The detection method according to any one of claims 1 to 4, wherein the separation column is a cation exchange resin column, preferably a strong acid type cation exchange resin column;
preferably, the column temperature of the separation column is 50-57 ℃, preferably 55 ℃.
6. The detection method according to any one of claims 1 to 5, wherein the derivatizing reagent used in the derivatization reaction is phthalic aldehyde, and preferably, the detection is detection of absorbance at a wavelength of 330 to 350 nm.
7. The detection method according to any one of claims 1 to 6, wherein the post-column derivatization reaction column is a quartz sand column, preferably, a column temperature of 30 to 50 ℃, further preferably 35 ℃.
8. The detection method according to any one of claims 1 to 7, wherein the mobile phase is one or two or more selected from the group consisting of a citric acid buffer solution, a trifluoroacetic acid solution, a sodium acetate solution, a mixture of sodium citrate and benzyl alcohol, and a mixture of ethanol and sodium hydroxide, and preferably the post-column derivatization buffer solution is one or two or three of a methanol solution of o-phthalaldehyde, a mixture of mercaptoethanol and a boric acid buffer solution, and an ethanol solution.
9. The detection method according to item 8, wherein gradient elution is performed, and the elution procedure is as follows:
firstly, a citric acid buffer solution is used as a mobile phase, and a post-column derivatization buffer solution is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and a boric acid buffer solution;
then, the mobile phase uses trifluoroacetic acid solution, and the post-column derivatization buffer solution is a mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses sodium acetate solution, and the post-column derivatization buffer solution is a mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses sodium acetate solution and the mixed solution of citric acid and benzyl alcohol, and the buffer solution for post-column derivatization is the mixed solution of phthalaldehyde methanol solution and mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses a mixed solution of citric acid and benzyl alcohol, and the post-column derivatization buffer solution is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses a mixed solution of ethanol and sodium hydroxide, and the buffer solution for post-column derivatization is an ethanol solution; and
finally, the mobile phase uses a citric acid buffer solution, and the post-column derivatization buffer solution is an ethanol solution.
10. The detection method according to any one of claims 1 to 9, wherein the amount of the sample is 10 to 20. Mu.L, preferably 15 to 20. Mu.L, and more preferably 20. Mu.L.
11. The detection method according to any one of claims 1 to 10, wherein the flow rate of the mobile phase is 0.35 to 0.55mL/min, preferably the flow rate of the post-column derivatization buffer solution is 0.2 to 0.4mL/min.
12. The method according to any one of claims 1 to 11, wherein the content of gamma-aminobutyric acid in the cosmetic is determined using an external standard method.
13. The method according to any one of items 1 to 12, wherein the detection limit of gamma-aminobutyric acid is not less than 3X 10 -4 ppm。
ADVANTAGEOUS EFFECTS OF INVENTION
The method for detecting the content of the gamma-aminobutyric acid in the cosmetics is simple to operate, sensitive in response, high in accuracy and good in repeatability, and can be used for detecting the content of the gamma-aminobutyric acid in the cosmetics.
According to the method, 0.02mol/L hydrochloric acid is used for heating in a water bath kettle at 60 ℃ for 4 hours, and gamma-aminobutyric acid in cosmetics is extracted. By using the separation column, the separation effect of gamma-aminobutyric acid can be greatly improved, the diffusion and overlapping phenomena of sample peaks are avoided, the peak shape repeatability of gamma-aminobutyric acid is good, and meanwhile, the accuracy of gamma-aminobutyric acid detection is improved. Phthalic dicarboxaldehyde as post-column derivatization reagent to reduce detection limit to 10 -4 ppm, can improve detection sensitivity and degree of accuracy greatly.
Drawings
FIG. 1 is a chromatogram of the control in example 1-1.
FIG. 2 is a chromatogram of the blank in example 1-1.
FIG. 3 is a chromatogram of the test sample in example 1-1.
Detailed Description
The present invention will now be described in detail with reference to the embodiments thereof as illustrated in the accompanying drawings, wherein like numerals refer to like features throughout. While specific embodiments of the invention are shown in the drawings, it should be understood that the invention may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
It should be noted that certain terms are used throughout the description and claims to refer to particular components. Those of skill in the art will understand that a person may refer to the same component by different names. The specification and claims do not identify differences in terms of components, but rather differences in terms of the functionality of the components. As referred to throughout the specification and claims, the terms "include" or "comprising" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. The description hereinafter sets forth a preferred embodiment for practicing the invention, but is not intended to limit the scope of the invention, as the description proceeds with reference to the general principles of the description. The scope of the invention is defined by the appended claims.
The gamma-aminobutyric acid is added into the cosmetic, so that the tight muscle nerve tissue can be relaxed, the skin can be penetrated quickly, the fine wrinkles can be reduced, the relaxing function of the muscle can be enhanced, and the effect of removing wrinkles can be achieved quickly.
Accordingly, the present invention provides a method for detecting gamma-aminobutyric acid in cosmetics, comprising the steps of:
preparation of a control: dissolving gamma-aminobutyric acid reference substance by using hydrochloric acid, and filtering to obtain gamma-aminobutyric acid reference substance solution;
preparation of test article: dissolving a cosmetic sample with hydrochloric acid, and filtering to obtain a sample solution;
preparation of a blank solution: filtering hydrochloric acid to obtain a blank solution;
and (3) measuring: detecting the reference substance solution, the test substance solution and the blank solution by using a separation column and a post-column derivatization reaction column, thereby obtaining the content of gamma-aminobutyric acid in the cosmetic test substance.
In one embodiment, the hydrochloric acid concentration is 0.02mol/L to 0.06mol/L, preferably 0.02mol/L.
For example, the concentration of the hydrochloric acid may be 0.02mol/L, 0.03mol/L, 0.04mol/L, 0.05mol/L, 0.06mol/L, etc.
The hydrochloric acid is prepared by diluting concentrated hydrochloric acid, for example, when the concentration of the prepared hydrochloric acid is 0.02mol/L, the preparation method is as follows: 1.67mL of concentrated hydrochloric acid was removed, added to pure water and the volume was set to 1000mL.
In one embodiment, during the preparation of the test sample, the dissolution is a water bath dissolution, preferably a dissolution time of 4 to 6 hours, further preferably a dissolution temperature of 60 to 80 ℃.
For example, the dissolution time may be 4 hours, 5 hours, or 6 hours, and the dissolution temperature may be 60 ℃, 65 ℃, 70 ℃, 75 ℃, 80 ℃, or the like.
The invention uses hydrochloric acid to dissolve the cosmetics, can effectively extract the gamma-aminobutyric acid in the cosmetics, has simple sample treatment and convenient operation, and reduces the loss of the gamma-aminobutyric acid in the samples.
In one embodiment, the cosmetic is a cream-like or liquid-like cosmetic, preferably, the cream-like cosmetic is an emulsion, an eye cream, or a facial cleanser; preferably, the liquid cosmetic is essential oil, toner or bath lotion.
In one embodiment, the separation column is a cation exchange resin column, preferably a strong acid cation exchange resin column, preferably a macroporous strong acid styrene cation exchange resin, more preferably a macroporous strong acid styrene cation exchange resin containing benzenesulfonic acid groups, further preferably a resin column having a filler particle size of 5 μm; preferably, the column temperature of the separation column is 50-57 ℃, preferably 55 ℃.
By using the separation column, the separation effect of gamma-aminobutyric acid can be greatly improved, the diffusion and overlapping phenomena of sample peaks are avoided, the peak shape repeatability of gamma-aminobutyric acid is good, and meanwhile, the accuracy of gamma-aminobutyric acid detection is improved.
For example, the column temperature of the separation column may be 50 ℃, 51 ℃, 52 ℃, 53 ℃, 54 ℃, 55 ℃, 56 ℃, 57 ℃, or the like.
In one embodiment, the derivatizing reagent used in the derivatization reaction is phthalic aldehyde, preferably, the detection is an absorbance at a detection wavelength of 330-350 nm.
For example, the detection wavelength may be 330nm, 335nm, 340nm, 345nm, 350nm, etc.
In one embodiment, the post-column derivatization reaction column is a quartz sand column, preferably at a column temperature of 30-50 ℃, further preferably 35 ℃.
Post-column derivatization refers to a technique that converts compounds into chemical structure-like substances using chemical transformations, which function to convert difficult-to-analyze substances into substances that are similar to their chemical structures but that are easy to analyze, facilitating quantification and separation. Chemical derivatization of amino acid analyzers refers to the chemical reaction of a sample component with a reagent (commonly known as a chemical derivatizing reagent or a labeling reagent) under conditions that result in a reaction product that facilitates chromatographic detection.
In one embodiment, the mobile phase is selected from one or more than two of citric acid buffer solution, trifluoroacetic acid solution, sodium acetate solution, mixed solution of citric acid and benzyl alcohol and mixed solution of ethanol and sodium hydroxide, preferably, the post-column derivatization buffer solution is one or two or three of phthalaldehyde methanol solution, mixed solution of mercaptoethanol and boric acid buffer solution and ethanol solution.
The citric acid buffer solution refers to a mixed solution of citric acid and an ethanol solution, preferably a mixed solution of 20wt% of citric acid and 11wt% of the ethanol solution;
the trifluoroacetic acid solution refers to a 2wt% trifluoroacetic acid solution;
the sodium acetate solution refers to 2.5wt% sodium acetate solution;
the mixed solution of the citric acid and the benzyl alcohol refers to a mixed solution of 6 weight percent of the citric acid and 0.5 weight percent of the benzyl alcohol;
the mixed solution of the ethanol and the sodium hydroxide refers to a mixed solution of 79wt% of ethanol and 0.8wt% of sodium hydroxide.
The phthalic aldehyde methanol solution refers to 2mg/mL of phthalic aldehyde methanol solution, and each 1mL of methanol solution contains 2mg of phthalic aldehyde;
the mixed solution of mercaptoethanol and boric acid buffer solution refers to mixed solution of 2mmol/L mercaptoethanol and 0.4mol/L boric acid buffer solution (pH=10-12);
the ethanol solution refers to a 50wt% ethanol solution.
Post-column derivatization refers to derivatization by mixing a test solution with a derivatizing agent in the presence of a buffer.
In one embodiment, a gradient elution is performed, with the elution procedure being:
firstly, a citric acid buffer solution is used as a mobile phase, and a post-column derivatization buffer solution is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and a boric acid buffer solution;
then, the mobile phase uses trifluoroacetic acid solution, and the post-column derivatization buffer solution is a mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses sodium acetate solution, and the post-column derivatization buffer solution is a mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses sodium acetate solution and the mixed solution of citric acid and benzyl alcohol, and the buffer solution for post-column derivatization is the mixed solution of phthalaldehyde methanol solution and mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses a mixed solution of citric acid and benzyl alcohol, and the post-column derivatization buffer solution is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and boric acid buffer solution;
then, the mobile phase uses a mixed solution of ethanol and sodium hydroxide, and the buffer solution for post-column derivatization is an ethanol solution; and
finally, the mobile phase uses a citric acid buffer solution, and the post-column derivatization buffer solution is an ethanol solution.
In one embodiment, the elution procedure is:
at 0min, the mobile phase uses citric acid buffer solution, and the post-column derivatization buffer solution is a mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
when the reaction time is 0-3.5min, the mobile phase uses trifluoroacetic acid solution, and the buffer solution for post-column derivatization is mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
3.5-6.5min, the mobile phase is sodium acetate solution, and the buffer solution for post-column derivatization is mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
6.5-12.8min, the mobile phase uses sodium acetate solution and the mixed solution of citric acid and benzyl alcohol, and the post-column derivatization buffer solution is the mixed solution of phthalaldehyde methanol solution and mercaptoethanol and boric acid buffer solution;
12.8-12.9min, the mobile phase uses the mixed solution of citric acid and benzyl alcohol, and the buffer solution for post-column derivatization is the mixed solution of phthalaldehyde methanol solution and mercaptoethanol and boric acid buffer solution;
12.9-21.0min, the mobile phase is mixed solution of ethanol and sodium hydroxide, and the buffer solution for post-column derivatization is ethanol solution; and
21.0-40.0min, the mobile phase is citric acid buffer solution, and the post-column derivatization buffer solution is ethanol solution.
Preferably, at 0min, the mobile phase uses a sodium citrate buffer, the citric acid buffer is a solution of 20wt% of citric acid and 11wt% of ethanol, the post-column derivatization buffer is a solution of phthalaldehyde methanol and a mixed solution of mercaptoethanol and boric acid buffer, the solution of phthalaldehyde methanol is a solution of 2mg/mL of phthalaldehyde methanol, the mixed solution of mercaptoethanol and boric acid buffer refers to a solution of 2mmol/L of mercaptoethanol+0.4 mol/L of boric acid buffer (pH=10), and the volume ratio of the solution of phthalaldehyde methanol to the mixed solution of mercaptoethanol and boric acid buffer is the same, and is 50%;
at 0-3.5min, the mobile phase is 2wt% trifluoroacetic acid solution, the post-column derivatization buffer solution is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and boric acid buffer solution, the phthalaldehyde methanol solution is a 2mg/mL phthalaldehyde methanol solution, the mixed solution of the mercaptoethanol and the boric acid buffer solution is 2mmol/L mercaptoethanol+0.4 mol/L boric acid buffer solution (pH=10), and the volume ratio of the phthalaldehyde methanol solution to the mixed solution of the mercaptoethanol and the boric acid buffer solution is the same and is 50%;
at 3.5-6.5min, 2.5wt% sodium acetate solution is used as a mobile phase, a buffer solution for post-column derivatization is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and boric acid buffer solution, wherein the phthalaldehyde methanol solution is a 2mg/mL phthalaldehyde methanol solution, the mixed solution of the mercaptoethanol and the boric acid buffer solution refers to a 2mmol/L mercaptoethanol+0.4mol/L boric acid buffer solution (pH=10), and the volume ratio of the phthalaldehyde methanol solution to the mixed solution of the mercaptoethanol and the boric acid buffer solution is the same and is 50%;
at the time of 6.5-12.8min, the mobile phase uses a sodium acetate solution and a mixed solution of citric acid and benzyl alcohol, wherein the sodium acetate solution is 2.5wt% of sodium acetate solution, the mixed solution of citric acid and benzyl alcohol refers to a mixed solution of 6wt% of citric acid and 0.5wt% of benzyl alcohol,
the post-column derivatization buffer solution is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and boric acid buffer solution, wherein the phthalaldehyde methanol solution is a 2mg/mL phthalaldehyde methanol solution, the mixed solution of the mercaptoethanol and the boric acid buffer solution refers to a 2mmol/L mercaptoethanol+0.4mol/L boric acid buffer solution (pH=10), and the volume ratios of the phthalaldehyde methanol solution and the mixed solution of the mercaptoethanol and the boric acid buffer solution are the same and are 50%;
wherein, at 6.5-6.6min, the mobile phase is a mixed solution of 23% sodium acetate solution and 77% citric acid and benzyl alcohol, and at 6.6-12.8min, the mobile phase is a mixed solution of 2% sodium acetate solution and 98% citric acid and benzyl alcohol;
12.8-12.9min, the mobile phase uses a mixed solution of citric acid and benzyl alcohol, wherein the mixed solution of citric acid and benzyl alcohol refers to a mixed solution of 6wt% of citric acid and 0.5wt% of benzyl alcohol,
the post-column derivatization buffer solution is a mixed solution of a phthalaldehyde methanol solution and a mercaptoethanol and boric acid buffer solution, wherein the phthalaldehyde methanol solution is a 2mg/mL phthalaldehyde methanol solution, the mixed solution of the mercaptoethanol and the boric acid buffer solution refers to a 2mmol/L mercaptoethanol+0.4mol/L boric acid buffer solution (pH=10), and the volume ratios of the phthalaldehyde methanol solution and the mixed solution of the mercaptoethanol and the boric acid buffer solution are the same and are 50%;
12.9-21.0min, the mobile phase uses a mixed solution of ethanol and sodium hydroxide, wherein the mixed solution of ethanol and sodium hydroxide refers to 79wt% of ethanol solution and 0.8wt% of sodium hydroxide solution;
the post-column derivatization buffer is an ethanol solution, and the ethanol solution is a 50wt% ethanol solution;
21.0-40.0min, the mobile phase uses citric acid buffer solution, the post-column derivatization buffer solution is ethanol solution, the sodium citrate buffer solution is 20wt% citric acid and 11wt% ethanol solution, and the ethanol solution is 50wt% ethanol solution.
In one embodiment, the amount of sample introduced is 10 to 20. Mu.L, preferably 15 to 20. Mu.L, and more preferably 20. Mu.L.
For example, the sample amount may be 10. Mu.L, 11. Mu.L, 12. Mu.L, 13. Mu.L, 14. Mu.L, 15. Mu.L, 16. Mu.L, 17. Mu.L, 18. Mu.L, 19. Mu.L, 20. Mu.L, etc.
In one embodiment, the flow rate of the mobile phase is from 0.35 to 0.55mL/min, and preferably, the flow rate of the post-column derivatizing buffer solution is from 0.2 to 0.4mL/min.
For example, the flow rate of the mobile phase may be 0.35mL/min, 0.40mL/min, 0.45mL/min, 0.50mL/min, 0.55mL/min, etc.;
the flow rate of the post-column derivatization buffer solution may be 0.20mL/min, 0.25mL/min, 0.30mL/min, 0.35mL/min, 0.40mL/min, and the like.
In one embodiment, the content of gamma-aminobutyric acid in the cosmetic is determined using an external standard method.
The external standard method is a quantitative method for estimating the concentration of a detected component in an unknown test material according to the functional relation between the response value of the control sample and the concentration of the standard substance added in the control sample. The external standard method used in the present invention is not limited, and a standard curve method may be used.
The detection method disclosed by the invention can be used for more accurately measuring the content of gamma-aminobutyric acid, and has the advantages of good repeatability and high accuracy.
In one embodiment, the detection accuracy of gamma-aminobutyric acid is not less than 3×10 -4 ppm。
The detection method adopts the hydrochloric acid water bath to extract the gamma-aminobutyric acid in the cosmetics, has simple and convenient operation, reduces the loss of the gamma-aminobutyric acid in the sample, and can reduce the detection limit by 10 by using the chromatographic conditions -4 ppm, the detection sensitivity and accuracy are greatly improved.
Examples
The materials used in the test and the test methods are described generally and/or specifically in the examples which follow,% represents wt%, i.e. weight percent, unless otherwise specified. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.
Example 1-1
1. Instrument reagent
LA8080 amino acid analyzer analytical balance: metretolidol AL104
Ultrapure water, hydrochloric acid (analytically pure) gamma-aminobutyric acid control (99.9%)
2. Chromatographic conditions
Separation column packed cation exchange resin column (4.4 mm. Times.60 mm,5 μm model: 852-8534)
Reaction column: filled quartz sand (4.6 mm. Times.40 mm, model: 8L 03600)
The separation flow rate is 0.45ml/min
Reaction flow rate: 0.3ml/min
Separation column temperature: 55 ℃;
reaction column temperature: 35 ℃;
sample injection amount: 20. Mu.L;
detection wavelength: 336nm
Gradient elution procedure:
| time/min | A％ | B％ | C％ | D％ | E％ | R1％ | R2％ | R3％ |
| 0.0 | 100 | -- | -- | -- | -- | 50 | 50 | -- |
| 3.5 | -- | 100 | -- | -- | -- | 50 | 50 | -- |
| 3.6 | -- | -- | 100 | -- | -- | 50 | 50 | -- |
| 6.5 | -- | -- | 100 | -- | -- | 50 | 50 | -- |
| 6.6 | -- | -- | 23 | 77 | -- | 50 | 50 | -- |
| 12.8 | -- | -- | 2 | 98 | -- | 50 | 50 | -- |
| 12.9 | -- | -- | -- | 100 | -- | 50 | 50 | -- |
| 21.0 | -- | -- | -- | -- | 100 | -- | -- | 100 |
| 32.1 | 100 | -- | -- | -- | -- | -- | 100 | |
| 40.0 | 100 | -- | -- | -- | -- | 100 |
Wherein, the mobile phase: a: 20wt% of citric acid and 11wt% of ethanol solution, and B: 2wt% of trifluoroacetic acid, C: sodium acetate solution 2.5wt%, D: citric acid 6wt%, 0.5wt% benzyl alcohol, E: 79wt% of ethanol solution and 0.8wt% of sodium hydroxide.
Buffer for derivatization reaction: r1:2mg/mL of a phthalic aldehyde methanol solution; r2:2mmol/L mercaptoethanol+0.4 mol/L boric acid buffer (ph=10); r3 is 50% ethanol solution.
The content is calculated using the following formula:
the calculation formula of the gamma-aminobutyric acid content in the test sample comprises the following steps:
note that: pi- -the content of gamma-aminobutyric acid in the test sample, mg/g
Cr-content of gamma-aminobutyric acid in reference substance solution, mg/ml
Ai- -peak area of sample solution
Peak area of Ar- -control solution
Vi- -volume of sample solution, ml
Mi- -weight of sample, g
Sample preparation:
control solution: precisely weighing gamma-aminobutyric acid reference substance 20mg to 100mL volumetric flask, metering volume to scale with 0.02mol/L hydrochloric acid, and mixing to obtain reference substance solution.
Test solution: weighing eye cream (Hua Xi organism, batch No. 210327) and adding into 10 g-10 mL volumetric flask, adding 0.02mol/L hydrochloric acid 9mL, extracting at 60deg.C for 4 hr, metering volume, and filtering to obtain sample solution.
Blank solution: the solution was filtered to obtain a blank solution containing 0.02mol/L hydrochloric acid.
4. Measurement
And (3) respectively taking a blank solution, a reference substance solution and a solution to be detected for sample injection, detecting according to the chromatographic conditions, and calculating the content of gamma-aminobutyric acid in the solution to be detected by using the peak area of an external standard method.
5. Results
The results are shown in Table 1, the chromatograms are shown in FIGS. 1-3, and the detection limit is 3×10 -4 ppm。
TABLE 1 detection results in example 1-1
Examples 1 to 2
The sample to be tested in example 1 was replaced with lot number: 210309 the content of gamma-aminobutyric acid in cosmetics was measured under the same conditions as in example 1, and the detection limit was 3X 10 as shown in Table 2 -4 ppm。
TABLE 2 detection results for examples 1-2
Examples 1 to 3
The detection wavelength in example 1 was changed to 350nm, and the detection limit was 7X 10 as shown in Table 3, except that the conditions were the same as in example 1, and the content of gamma-aminobutyric acid in the cosmetic was the same -4 ppm。
TABLE 3 detection results for examples 1-3
Examples 1 to 4
The content of gamma-aminobutyric acid in cosmetics was measured under the same conditions as in example 1 except that the flow rate of the mobile phase in example 1 was changed to 0.55ml/min, and the detection limit was 9X 10 as shown in Table 4 -4 ppm。
TABLE 4 detection results for examples 1-4
Examples 1 to 5
The flow rate of the post-column-derived buffer solution in example 1 was changed to 0.4ml/min, and the content of gamma-aminobutyric acid in the cosmetic was measured under the same conditions as in example 1, and the detection limit was 6.6X10 as shown in Table 5 -4 ppm。
TABLE 5 chromatographic analysis results of samples to be tested of examples 1-5
Examples 1 to 6
The content of gamma-aminobutyric acid in cosmetics was measured under the same conditions as in example 1 except that the flow rate of the mobile phase in example 1 was changed to 0.35ml/min, and the detection limit was 10X 10 as shown in Table 6 -4 ppm。
TABLE 6 detection results for examples 1-6
Examples 1 to 7
The flow rate of the post-column-derived buffer solution in example 1 was changed to 0.2ml/min, and the content of gamma-aminobutyric acid in the cosmetic was measured under the same conditions as in example 1, and the detection limit was 13.5X10 as shown in Table 7 -4 ppm。
TABLE 7 detection results for examples 1-7
Examples 1 to 8
The sample volume in example 1 was changed to 10. Mu.L, and the content of gamma-aminobutyric acid in the cosmetic was measured under the same conditions as in example 1, and the detection limit was 5.4X10 as shown in Table 8 -4 ppm。
TABLE 8 detection results for examples 1-8
Examples 1 to 9
The sample volume in example 1 was changed to 15. Mu.L, and the content of gamma-aminobutyric acid in the cosmetic was measured under the same conditions as in example 1, and the detection limit was 7.8X10 as shown in Table 9 -4 ppm。
TABLE 9 detection results for examples 1-9
Examples 1 to 10
The measurement of the content of gamma-aminobutyric acid in cosmetics was carried out under the same conditions as in example 1 except that the dissolution time of the sample preparation in example 1 was changed to 6 hours, and the detection limit was 3.2X10 as shown in Table 10 -4 ppm。
TABLE 10 detection results for examples 1-10
Examples 1 to 11
The content of gamma-aminobutyric acid in cosmetics was measured by changing the dissolution temperature of the sample preparation in example 1 to 80℃under the same conditions as in example 1, and the detection limit was 3.1X10 as shown in Table 11 -4 ppm。
TABLE 11 detection results for examples 1-11
Examples 1 to 12
The column temperature of the separation column in example 1 was changed to 50℃and the content of gamma-aminobutyric acid in the cosmetic was measured under the same conditions as in example 1, and the detection limit was 4.3X10 as shown in Table 12 -4 ppm。
TABLE 12 detection results for examples 1-12
Examples 1 to 13
The column temperature of the separation column in example 1 was changed to 57℃and the content of gamma-aminobutyric acid in the cosmetic was measured under the same conditions as in example 1, and the detection limit was 5.8X10 as shown in Table 13 -4 ppm。
TABLE 13 detection results for examples 1-13
Examples 1 to 14
The measurement of the content of gamma-aminobutyric acid in cosmetics was carried out under the same conditions as in example 1 except that the detection wavelength in example 1 was changed to 330nm, and the detection limit was 3.9X10 as shown in Table 14 -4 ppm。
TABLE 14 detection results for examples 1-14
TABLE 15 chromatographic conditions, extraction conditions and detection limits used in the examples
Example 2-1 determination of repeatability
According to the sample treatment method in example 1-1, 6 parts of the same lot number sample as in example 1-1 was prepared in parallel, and the control solution and the detection conditions were the same as in example 1-1, and the measurement results are shown in Table 16.
TABLE 16 results of the content of gamma-aminobutyric acid of example 2-1
Example 2-measurement of precision between days
According to the sample treatment method in example 1-1, 6 samples of the same lot as in example 1-1 were prepared in parallel at different times, and the control solution and the detection conditions were the same as in example 1-1, using different instruments, and the measurement results are shown in Table 17.
TABLE 17 results of the content of gamma-aminobutyric acid of examples 2-2
Examples 2-3 determination of accuracy
Taking a blank solution of gamma-aminobutyric acid, treating the blank solution according to the treatment mode of the test sample in the embodiment 1-1, precisely weighing the reference substance in the embodiment 1-1, and preparing 3 parts of test sample solutions with concentration of 80%, 100% and 120% of the prescription amount by a simulated prescription in parallel, wherein the total amount is 9 parts. The control solution and the detection conditions were the same as in example 1-1, and the measurement results are shown in Table 18.
Table 18 gamma-aminobutyric acid accuracy validation results
Comparative example 1
The content of gamma-aminobutyric acid in cosmetics was measured by changing the phthalic dicarboxaldehyde in example 1 to ninhydrin under the same conditions as in example 1, and the detection limit was 500X 10 as shown in Table 19 -4 ppm。
Table 19 results of comparative example 1
As can be seen from the above table, the data parallelism is poor.
Comparative example 2
The content of gamma-aminobutyric acid in cosmetics was measured by changing the temperature of the reaction column in example 1 to 135℃under the same conditions as in example 1, and the detection limit was 70X 10 as shown in Table 20 -4 ppm。
Table 20 results of comparative example 2
As can be seen from the above table, increasing the temperature of the reaction column may affect the progress of the derivatization reaction, resulting in a larger deviation of the results.
Comparative example 3
The extraction method of gamma-aminobutyric acid in example 1 was changed to 9ml of water, and after 4 hours of extraction at 60 ℃, the volume was fixed and filteredThe conditions were the same as in example 1, and the content of gamma-aminobutyric acid in the cosmetic was measured, and the results are shown in Table 21, and the detection limit was 100X 10 -4 ppm。
Table 21 results of comparative example 3
From the above table, it can be seen that water cannot completely extract gamma-aminobutyric acid in cosmetics, resulting in a smaller result.
Comparative example 4
The method of extracting gamma-aminobutyric acid in example 1 was changed to 9ml of 0.02mol/L hydrochloric acid, ultrasonic extraction was carried out for 4 hours, the volume was determined, filtration was carried out, the other conditions were the same as in example 1, and the content of gamma-aminobutyric acid in cosmetics was measured, and the detection limit was 135×10 as shown in Table 22 -4 ppm。
Table 22 results of comparative example 4
From the above table, it can be seen that ultrasonic extraction for 4 hours cannot completely extract gamma-aminobutyric acid in cosmetics, resulting in a smaller result.
Comparative example 5
The extraction mode of gamma-aminobutyric acid in example 1 was changed to: petroleum ether dissolved sample, 0.02mol/L hydrochloric acid extraction, taking the lower layer liquid to constant volume, filtering, and measuring gamma-aminobutyric acid content in cosmetics under the same conditions as in example 1, wherein the detection limit is 309×10 as shown in Table 23 -4 ppm。
Table 23 results of comparative example 5
As can be seen from the above table, the parallelism of the results is poor, and the extraction mode operation is complicated.
Comparative example 6
Gradient elution was performed on A, B, C, D, E, R1, R2 and R3 in example 1, and the elution ratio and time were changed to those shown in table 24:
TABLE 24 elution procedure
| Time/min | A％ | B％ | C％ | D％ | E％ | R1％ | R2％ | R3％ |
| 0.0 | 100 | -- | -- | -- | -- | 50 | 50 | -- |
| 3.0 | 100 | 100 | -- | -- | -- | 50 | 50 | -- |
| 3.1 | -- | 100 | -- | -- | -- | 50 | 50 | -- |
| 5.4 | -- | -- | 100 | -- | -- | 50 | 50 | -- |
| 5.5 | -- | -- | 100 | -- | -- | 50 | 50 | -- |
| 13.8 | -- | -- | -- | 100 | -- | 50 | 50 | -- |
| 13.9 | -- | -- | -- | 100 | -- | 50 | 50 | -- |
| 30.0 | -- | -- | -- | -- | 100 | -- | -- | -- |
| 30.1 | -- | -- | -- | -- | 100 | -- | -- | -- |
| 32.0 | -- | -- | -- | -- | 100 | -- | -- | -- |
| 32.1 | -- | -- | -- | -- | 100 | -- | -- | 100 |
| 33.0 | -- | -- | -- | -- | -- | -- | -- | 100 |
| 33.1 | -- | 100 | -- | -- | -- | -- | -- | 100 |
| 34.0 | -- | 100 | -- | -- | -- | -- | -- | 100 |
| 34.1 | 100 | -- | -- | -- | -- | -- | -- | 100 |
| 37.0 | 100 | -- | -- | -- | -- | -- | -- | 100 |
| 37.1 | 100 | -- | -- | -- | -- | 50 | 50 | -- |
| 53.0 | 100 | -- | -- | -- | -- | 50 | 50 | -- |
The other test conditions were the same as in example 1, and the content of gamma-aminobutyric acid in the cosmetic was measured, and the results are shown in Table 25.
Table 25 results of comparative example 6
As can be seen from the table, the elution mode is the gradient elution condition of the common amino acid analyzer, the peak time of gamma-aminobutyric acid is 1.887min, the gamma-aminobutyric acid is severely interfered by a solvent peak, the parallelism is poor, and the detection limit is high.
Comparative example 7
In example 1, the mobile phases B and C used were changed to B: 22wt% of citric acid solution, C: 1.3% by weight of sodium citrate solution, the remainder was the same as in example 1, and the content of gamma-aminobutyric acid in the cosmetic was measured, and the results are shown in Table 26, with a detection limit of 240X 10 -4 ppm。
Table 26 results of comparative example 7
From the above table, the citric acid and sodium citrate solution have poor eluting ability to gamma-aminobutyric acid, which results in low peak area of gamma-aminobutyric acid, smaller result and high detection limit.
Comparative example 8
The derivatization reagent buffer mercaptoethanol in example 1 was changed to mercaptopropionic acid, and the other conditions were the same as in example 1, and no peak of gamma-aminobutyric acid in the sample was found as a result of detection, and the detection limit was 0.
Comparative example 9
The detection wavelength in example 1 was changed to 570nm, and the detection result was not found to be a peak of gamma-aminobutyric acid in the sample, and the detection limit was 0, under the same conditions as in example 1.
Table 27 chromatographic conditions, extraction conditions and detection limits used in comparative examples
The above description is only a preferred embodiment of the present invention, and is not intended to limit the invention in any way, and any person skilled in the art may make modifications or alterations to the disclosed technical content to the equivalent embodiments. However, any simple modification, equivalent variation and variation of the above embodiments according to the technical substance of the present invention still fall within the protection scope of the technical solution of the present invention.
Claims (19)
1. A method for detecting gamma-aminobutyric acid in cosmetics, which comprises the following steps:
preparation of a control: dissolving gamma-aminobutyric acid reference substance by using hydrochloric acid, and filtering to obtain gamma-aminobutyric acid reference substance solution;
preparation of test article: dissolving a cosmetic sample with hydrochloric acid, and filtering to obtain a sample solution;
preparation of a blank solution: filtering hydrochloric acid to obtain a blank solution;
and (3) measuring: detecting the reference substance solution, the test substance solution and the blank solution by using a separation column and a post-column derivatization reaction column, thereby obtaining the content of gamma-aminobutyric acid in the cosmetic test substance;
the elution was performed using a gradient elution procedure:
at 0min, the mobile phase uses citric acid buffer solution, and the post-column derivatization buffer solution is a mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
when the reaction time is 0-3.5min, the mobile phase uses trifluoroacetic acid solution, and the buffer solution for post-column derivatization is mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
3.5-6.5min, the mobile phase is sodium acetate solution, and the buffer solution for post-column derivatization is mixed solution of o-phthalaldehyde methanol solution, mercaptoethanol and boric acid buffer solution;
6.5-12.8min, the mobile phase uses sodium acetate solution and the mixed solution of citric acid and benzyl alcohol, and the post-column derivatization buffer solution is the mixed solution of phthalaldehyde methanol solution and mercaptoethanol and boric acid buffer solution;
12.8-12.9min, the mobile phase uses the mixed solution of citric acid and benzyl alcohol, and the buffer solution for post-column derivatization is the mixed solution of phthalaldehyde methanol solution and mercaptoethanol and boric acid buffer solution;
12.9-21.0min, the mobile phase is mixed solution of ethanol and sodium hydroxide, and the buffer solution for post-column derivatization is ethanol solution; and
21.0-40.0min, using citric acid buffer solution as mobile phase, and using ethanol solution as post-column derivatization buffer solution;
the separation column is a cation exchange resin column;
the post-column derivatization reaction column is a quartz sand column;
the column temperature of the reaction column is 30-50 ℃;
the detection is to detect the absorbance with the wavelength of 330-350 nm;
the derivatization reagent used in the derivatization reaction is phthalic dicarboxaldehyde.
2. The detection method according to claim 1, wherein the concentration of the hydrochloric acid is 0.02mol/L to 0.06mol/L.
3. The method of claim 1, wherein the dissolving is a water bath dissolving.
4. The method according to claim 1, wherein the dissolution time is 4 to 6 hours.
5. The method according to claim 1, wherein the dissolution temperature is 60-80 ℃.
6. The detection method according to claim 1, wherein the cosmetic is a cream-like or liquid-like cosmetic.
7. The detection method according to claim 6, wherein the cream-milky cosmetic is an emulsion, an eye cream, or a facial cleanser.
8. The detection method according to claim 6, wherein the liquid cosmetic is essential oil, toner or shower gel.
9. The detection method according to claim 1, wherein the separation column is a strong acid type cation exchange resin column.
10. The detection method according to claim 1, wherein the column temperature of the separation column is 50-57 ℃.
11. The detection method according to claim 1, wherein the column temperature of the separation column is 55 ℃.
12. The detection method according to claim 1, wherein the column temperature of the reaction column is 35 ℃.
13. The detection method according to any one of claims 1 to 12, wherein the amount of sample introduced is 10 to 20 μl.
14. The detection method according to any one of claims 1 to 12, wherein the amount of sample introduced is 15 to 20 μl.
15. The detection method according to any one of claims 1 to 12, wherein the amount of sample introduced is 20 μl.
16. The detection method according to any one of claims 1 to 12, wherein the flow rate of the mobile phase is 0.35 to 0.55mL/min.
17. The assay of any one of claims 1-12, wherein the flow rate of post-column derivatization buffer solution is 0.2-0.4mL/min.
18. The method according to any one of claims 1 to 12, wherein the content of gamma-aminobutyric acid in the cosmetic is determined using an external standard method.
19. The method according to any one of claims 1 to 12, wherein the limit of detection of gamma-aminobutyric acid is not less than 3X 10 - 4 ppm。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110780174.4A CN113341029B (en) | 2021-07-09 | 2021-07-09 | Method for detecting content of gamma-aminobutyric acid in cosmetics |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110780174.4A CN113341029B (en) | 2021-07-09 | 2021-07-09 | Method for detecting content of gamma-aminobutyric acid in cosmetics |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113341029A CN113341029A (en) | 2021-09-03 |
| CN113341029B true CN113341029B (en) | 2023-08-18 |
Family
ID=77479455
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110780174.4A Active CN113341029B (en) | 2021-07-09 | 2021-07-09 | Method for detecting content of gamma-aminobutyric acid in cosmetics |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113341029B (en) |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0233973A1 (en) * | 1986-02-26 | 1987-09-02 | Hewlett-Packard GmbH | Mixture of amino acid derivatives, process of producing the mixture and use of the mixture for quantitative determination of the amino acids |
| JPH03199944A (en) * | 1989-12-27 | 1991-08-30 | Shimadzu Corp | Analyzing method for amino compound |
| JP2004045174A (en) * | 2002-07-11 | 2004-02-12 | Yakult Honsha Co Ltd | Amino acid analyzing method |
| CN101480216A (en) * | 2008-01-11 | 2009-07-15 | 北京健健康康生物技术有限公司 | Method for extracting composite aminoacid from water melon or black seed melon |
| CN102230921A (en) * | 2011-03-29 | 2011-11-02 | 宁波谱尼测试技术有限公司 | Detection method for gamma-aminobutyric acid content in food by using high performance liquid chromatography method |
| CN103713077A (en) * | 2013-12-26 | 2014-04-09 | 晨光生物科技集团股份有限公司 | Method for determining content of gamma-aminobutyric acid in red yeast through high-efficient liquid chromatography |
| CN105651874A (en) * | 2015-12-30 | 2016-06-08 | 河北三元食品有限公司 | Special-purpose high-precision high-performance liquid chromatography method for detecting gamma-aminobutyric acid and sixteen amino acids hydrolyzed from protein in milk-including drink at the same time |
| CN109115917A (en) * | 2018-10-16 | 2019-01-01 | 华熙福瑞达生物医药有限公司 | A kind of measuring method of alpha-aminobutyric acid content |
| CN109959736A (en) * | 2019-04-09 | 2019-07-02 | 北京市药品检验所 | Use the method for amino-acid analyzer measurement tranexamic acid content |
| CN111007034A (en) * | 2019-12-16 | 2020-04-14 | 贵州医科大学 | Method for detecting content of amino acid in hereditary metabolic disease |
| CN111323503A (en) * | 2020-03-12 | 2020-06-23 | 河北科星药业有限公司 | Method for measuring content of compound amino acid injection |
| CN112602906A (en) * | 2020-12-21 | 2021-04-06 | 河南省科学院高新技术研究中心 | Processing method for improving content of gamma-aminobutyric acid in rhizome food |
| CN113030342A (en) * | 2021-04-14 | 2021-06-25 | 华熙生物科技股份有限公司 | Method for detecting glutamic acid residue in gamma-aminobutyric acid |
-
2021
- 2021-07-09 CN CN202110780174.4A patent/CN113341029B/en active Active
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0233973A1 (en) * | 1986-02-26 | 1987-09-02 | Hewlett-Packard GmbH | Mixture of amino acid derivatives, process of producing the mixture and use of the mixture for quantitative determination of the amino acids |
| JPH03199944A (en) * | 1989-12-27 | 1991-08-30 | Shimadzu Corp | Analyzing method for amino compound |
| JP2004045174A (en) * | 2002-07-11 | 2004-02-12 | Yakult Honsha Co Ltd | Amino acid analyzing method |
| CN101480216A (en) * | 2008-01-11 | 2009-07-15 | 北京健健康康生物技术有限公司 | Method for extracting composite aminoacid from water melon or black seed melon |
| CN102230921A (en) * | 2011-03-29 | 2011-11-02 | 宁波谱尼测试技术有限公司 | Detection method for gamma-aminobutyric acid content in food by using high performance liquid chromatography method |
| CN103713077A (en) * | 2013-12-26 | 2014-04-09 | 晨光生物科技集团股份有限公司 | Method for determining content of gamma-aminobutyric acid in red yeast through high-efficient liquid chromatography |
| CN105651874A (en) * | 2015-12-30 | 2016-06-08 | 河北三元食品有限公司 | Special-purpose high-precision high-performance liquid chromatography method for detecting gamma-aminobutyric acid and sixteen amino acids hydrolyzed from protein in milk-including drink at the same time |
| CN109115917A (en) * | 2018-10-16 | 2019-01-01 | 华熙福瑞达生物医药有限公司 | A kind of measuring method of alpha-aminobutyric acid content |
| CN109959736A (en) * | 2019-04-09 | 2019-07-02 | 北京市药品检验所 | Use the method for amino-acid analyzer measurement tranexamic acid content |
| CN111007034A (en) * | 2019-12-16 | 2020-04-14 | 贵州医科大学 | Method for detecting content of amino acid in hereditary metabolic disease |
| CN111323503A (en) * | 2020-03-12 | 2020-06-23 | 河北科星药业有限公司 | Method for measuring content of compound amino acid injection |
| CN112602906A (en) * | 2020-12-21 | 2021-04-06 | 河南省科学院高新技术研究中心 | Processing method for improving content of gamma-aminobutyric acid in rhizome food |
| CN113030342A (en) * | 2021-04-14 | 2021-06-25 | 华熙生物科技股份有限公司 | Method for detecting glutamic acid residue in gamma-aminobutyric acid |
Non-Patent Citations (1)
| Title |
|---|
| 离子交换色谱-茚三酮柱后衍生法同时测定绿茶饮料中39种游离氨基酸;贺习文;高勤叶;黄姣姣;李易轩;张兴悟;马川;;饮料工业(第01期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113341029A (en) | 2021-09-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102520079B (en) | Method for rapidly measuring content of solanesol in tobaccos by using UPLC (Ultra Performance Liquid Chromatography) | |
| CN113433257B (en) | Method for separating and measuring carnitine enantiomer in health food | |
| CN102539568A (en) | Ion chromatography method for detecting iodate and bromate in cosmetic | |
| CN113267588B (en) | Method for simultaneously detecting 7 acrylic acid and acrylic ester monomers in plastic product | |
| CN113341029B (en) | Method for detecting content of gamma-aminobutyric acid in cosmetics | |
| CN102955008B (en) | Method for detecting sulfonamide residue in eel by pressurized capillary electrochromatography | |
| CN109490444A (en) | A kind of method of degradation impurity in HPLC method separation determination dutasteride bulk pharmaceutical chemicals and preparation | |
| CN106198811B (en) | Quickly measure the kit and method of water and aquatic products Malachite Green and crystal violet | |
| CN102818782B (en) | Determination method of total 2-(2-phenethyl) chromone compound content | |
| CN109709222B (en) | Component detection method of Ganmaoling and compound Ganmaoling | |
| CN111965273A (en) | HPLC (high Performance liquid chromatography) method for detecting genotoxic impurities in candesartan cilexetil | |
| CN107266627B (en) | A kind of the core-shell type molecularly imprinted polymer and preparation method of recognizable erythrosine | |
| CN115389666A (en) | Method for efficiently and simultaneously detecting ergothioneine and ectoine in cosmetics | |
| CN110672768B (en) | Method for determining diethoxydiol in oral care product | |
| CN109030668B (en) | High performance liquid phase analysis method of gadoxetic acid disodium intermediate | |
| CN113740198A (en) | Method for determining yellow vaseline content in Mayinglong musk hemorrhoid ointment | |
| CN108226349B (en) | Detection method for simultaneously determining multiple bacteriostatic agents in tramadol cream | |
| CN114216972A (en) | Method for measuring content of dipalmitoyl hydroxyproline | |
| CN115128177A (en) | Method for analyzing and determining genotoxic impurities in ganciclovir condensation compound by using HPLC method | |
| CN103048407A (en) | Content detection method for lysine of compound ketoacid tablet | |
| CN109387588B (en) | Separation method of water-soluble ultraviolet absorbent and application thereof | |
| CN107807182B (en) | Method for measuring content of ganoderic acid A in ganoderma lucidum syrup | |
| CN101625340A (en) | High performance liquid chromatogram detecting method for detecting metaphenylene diamine in cosmetic oxidizing-type hair colorant | |
| CN104965034A (en) | Detection method for rhododendrol in cosmetics | |
| Cruz-Vera et al. | Fast urinary screening for imipramine and desipramine using on-line solid-phase extraction and selective derivatization |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20230613 Address after: Tianchen Avenue, Ji'nan hi tech Development Zone of Shandong Province, No. 678 250101 Applicant after: BLOOMAGE BIOTECH Co.,Ltd. Address before: Tianchen Avenue, Ji'nan hi tech Development Zone of Shandong Province, No. 678 250101 Applicant before: BLOOMAGE BIOTECH Co.,Ltd. Applicant before: SHANDONG BLOOMAGE HYINC BIOPHARM Corp.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |