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HPLC (high performance liquid chromatography) measuring method for amino acid of hydrolyzed vegetable protein (HVP) liquid

A technology of hydrolyzed vegetable protein and determination method, which is applied in the field of HPLC determination of amino acids in hydrolyzed vegetable protein liquid, which can solve the problems of poor separation effect of natural hydrolyzed amino acid, strong irritation of tetrahydrofuran, harm to human and environment, etc., and achieve analysis accuracy High, low matrix interference, good separation effect

Inactive Publication Date: 2017-01-25
LIAOCHENG XINHENGJI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

2,4-Dinitrofluorobenzene reacts with amino acids to generate dinitrophenol amino acid derivatives, but it uses a fluorescence detector, and tetrahydrofuran is added to the mobile phase. Tetrahydrofuran is very irritating and has a strong smell. It is harmful to people and the environment. Secondly, it uses a first-order gradient elution, which is not very effective for the separation of complex natural hydrolyzed amino acids.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The HPLC assay method of amino acid in the hydrolyzed vegetable protein liquid, comprises the steps:

[0037] (1) Preparation of derivatization buffer, equilibration buffer and derivatization reagent:

[0038] Derivative buffer solution: Weigh 1.300g of boric acid and 7.532g of borax, dissolve in water, transfer to a 500ml volumetric flask, dilute to volume, and shake well.

[0039] Equilibrium buffer solution: Weigh 0.910g of potassium dihydrogen phosphate and 3.580g of disodium hydrogen phosphate dodecahydrate, add water to dissolve, transfer to a 500ml volumetric flask, dilute to volume, and shake well.

[0040] Derivatization reagent: 1% (volume fraction) 2,4-dinitrofluorobenzene in acetonitrile solution.

[0041] (2) HVP liquid sample solution: Accurately weigh 0.252g of HVP liquid, and dilute it in a 100ml volumetric flask. Take 5.0ml of HVP solution, add 5ml of derivatization buffer to a 50ml brown volumetric flask, mix well, add 5.0ml of derivatization reagent...

Embodiment 2

[0049] The HPLC assay method of amino acid in the hydrolyzed vegetable protein liquid, comprises the steps:

[0050] (1) Preparation of derivatization buffer, equilibration buffer and derivatization reagent:

[0051] Derivative buffer solution: Weigh 1.240g of boric acid and 7.630g of borax, add water to dissolve, transfer to a 500ml volumetric flask, dilute to volume, and shake well.

[0052] Equilibrium buffer solution: Weigh 0.980g of potassium dihydrogen phosphate and 3.475g of disodium hydrogen phosphate dodecahydrate, add water to dissolve, transfer to a 500ml volumetric flask, dilute to volume, and shake well.

[0053] Derivatization reagent: 1% (volume fraction) 2,4-dinitrofluorobenzene in acetonitrile solution.

[0054] (2) HVP liquid sample solution: Accurately weigh 0.252g of HVP liquid, and dilute it in a 100ml volumetric flask. Take 5.0ml of HVP solution, add 5ml of derivatization buffer to a 50ml brown volumetric flask, mix well, add 5.0ml of derivatization reage...

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Abstract

The invention relates to an HPLC (high performance liquid chromatography) measuring method for amino acid of hydrolyzed vegetable protein (HVP) liquid. According to the method, a salt solution serves as a flowing phase on the basis of HPLC, and certain N,N-dimethyl formamide and acetic acid are added to improve chromatographic peak shape and separation degree. The amino acid of the HVP liquid is measured by a 2,4-dinitrofluorobenzene pre-column derivation-HPLC-ultraviolet detection method. The peak shape is improved by preparing and taking the specific salt solution as the flowing phase, so that the separation degree of each amino acid derivative is good, the matrix interference is small and quantification is more accurate. The method is high in analysis precision, is particularly suitable for various amino acid mixtures and is good in separation and accurate in quantification.

Description

technical field [0001] The invention relates to an HPLC assay method for amino acids in hydrolyzed vegetable protein liquid, and belongs to the technical field of analysis and detection. Background technique [0002] HVP is hydrolyzed vegetable protein, which refers to the product of vegetable protein hydrolysis under the action of acid catalysis. Its constituents are mainly amino acids, so it is also called amino acid liquid. [0003] At present, the analysis methods of amino acids mainly include chemical analysis, electrochemical methods, spectrophotometry, high performance liquid chromatography (HPLC), capillary electrophoresis, gas chromatography, etc. These analysis methods can be very good for simple amino acid components. However, for the analysis of a mixture of various complex amino acids, the effect is not very good, the quantification is not accurate and the deviation is large due to factors such as poor resolution and matrix interference. [0004] At present, t...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/74G01N30/34
CPCG01N30/02G01N30/34G01N30/74
Inventor 张海燕张甲贵
Owner LIAOCHENG XINHENGJI BIOTECH
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