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Method for determining content of gamma-aminobutyric acid in red yeast through high-efficient liquid chromatography

A high-performance liquid chromatography, aminobutyric acid technology, applied in the field of determination of red yeast yeast components, can solve the problems of low accuracy of detection results, unstable and reliable, interference detection effect, etc., to improve detection accuracy, reduce interference, Less interference from impurities

Active Publication Date: 2014-04-09
杭州天曲生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complex composition and many impurities in red yeast rice products, there are many factors that interfere with the detection effect, resulting in low accuracy and large deviation of the detection results, which are not stable and reliable

Method used

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  • Method for determining content of gamma-aminobutyric acid in red yeast through high-efficient liquid chromatography
  • Method for determining content of gamma-aminobutyric acid in red yeast through high-efficient liquid chromatography
  • Method for determining content of gamma-aminobutyric acid in red yeast through high-efficient liquid chromatography

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: The method for measuring the content of gamma-aminobutyric acid in red yeast rice by this high-performance liquid chromatography adopts the following raw materials, instruments and process steps.

[0026] 1. The experimental equipment and reagents are as follows

[0027] 1) Instrument: Agilent 1260 high performance liquid chromatography (Agilent, USA); chromatographic column is Diamonsil C18 250 x 4.6 mm, 5 μm (Dima Technology); syringe microporous membrane; 0.22 μm organic filter membrane; spin Evaporator (Shanghai Shenke); electronic balance (Mettler-Toledo Instrument Co., Ltd.); high-speed centrifuge (Xiangyi Group); ultrasonic instrument.

[0028] 2) Experimental reagents: γ-aminobutyric acid standard, methanol, acetonitrile, formic acid, tetrahydrofuran, ultrapure water.

[0029] Second, the experiment is as follows:

[0030] 1) Experiment with γ-amino acid standard and determine the best chromatographic conditions.

[0031] 2) Use the established ...

Embodiment 2

[0066] Embodiment 2: The method for the determination of gamma-aminobutyric acid content in red yeast rice by this high performance liquid chromatography except the following difference, other is the same as embodiment 1.

[0067] (1) Sample handling:

[0068] (1) Extraction of γ-aminobutyric acid from red yeast rice samples: Accurately weigh 2.0g of red yeast rice samples and put them in a 100mL centrifuge tube; add 60ml of ultrapure water according to the mass-volume ratio of 1:30, and ultrasonically extract for 2.5 hours. Centrifuge at 5000rmp for 8 minutes to collect the supernatant; repeat the above steps 5 times, and combine the supernatants; add 1 times the volume of absolute ethanol and 1 / 10 volume of 7wt% ammonium sulfate solution, and precipitate at 0°C for 10 minutes;

[0069] (2) Centrifuge at 5000rmp for 8 minutes, discard the precipitate, concentrate under reduced pressure at room temperature to remove the solvent, then add the compound enzyme preparation, adjust...

Embodiment 3

[0074] Embodiment 3: The method for the determination of gamma-aminobutyric acid content in red yeast rice by this high performance liquid chromatography except the following difference, other is the same as embodiment 1.

[0075] (1) Sample handling:

[0076] (1) Extraction of γ-aminobutyric acid from red yeast rice samples: Accurately weigh 2.0g of red yeast rice samples and put them in a 50mL centrifuge tube; add 20ml of ultrapure water according to the mass-volume ratio of 1:10, and ultrasonically extract for 4 hours. Centrifuge at 7000rmp for 5min, collect the supernatant; repeat the above steps 4 times, combine the supernatant; add 1.5 times the volume of absolute ethanol and 1 / 5 volume of 5wt% potassium acetate solution, and precipitate at 4°C for 20min;

[0077] (2) Centrifuge at 7000rmp for 5min, discard the precipitate, concentrate under reduced pressure at room temperature to remove the solvent, then add the compound enzyme preparation, adjust the pH=3.5, and treat ...

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Abstract

The invention discloses a method for determining the content of gamma-aminobutyric acid in red yeast through a high-efficient liquid chromatography. The method comprises the steps of (1) extracting the gamma-aminobutyric acid: adding the measured red yeast sample into water, ultrasonically extracting, centrifuging, and collecting the supernatant; (2) precipitating alcohol: adding ethanol and salt solution into the supernatant to execute alcohol precipitation, centrifuging the solution, and collecting the alcohol precipitation supernatant; (3) carrying out enzyme treatment: separating the solvent from the alcohol precipitation supernatant, and adding a composite enzymic preparation to execute enzyme treatment; (4) carrying out depigmentation: adsorbing the enzyme treated concentrated solution onto a column, adsorbing the solution with resin, eluting the solution with ammonia water; filtering the elution solution by utilizing a micro-pore filter membrane, wherein the filtered liquid is the detected liquid; (5) executing pre-column derivatization on the detected liquid, i.e. determining the content of the gamma-aminobutyric acid by adopting the liquid chromatography. By adopting the steps of alcohol precipitation, enzyme treatment and depigmentation, the content of impurities in the gamma-aminobutyric acid can be effectively removed, the interference in the detection process can be reduced, the detection accuracy can be effectively improved, and the detection result is reliable and stable.

Description

technical field [0001] The invention relates to a method for determining the components of red yeast rice, in particular to a method for determining the content of gamma-aminobutyric acid in red yeast rice by high performance liquid chromatography. Background technique [0002] γ-aminobutyric acid (γ-aminobuyric acid, GABA) is a physiologically active non-protein amino acid widely present in animals and plants. It has various physiological functions such as regulating cardiovascular activity and arrhythmia, and nourishing nerve cells. With the deepening of research, GABA has developed into a new type of functional factor and is gradually used in medicine, food and feed industries. Monascus sp. is a traditional food fermentation microorganism in my country, which can produce rich enzymes, pigments and various physiologically active substances, and has been widely used in food coloring, brewing, medical care and other aspects. Animal and clinical tests have shown that the fer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 王轩武亚明李静静
Owner 杭州天曲生物技术有限公司
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