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Method for determining content of beta-hydroxy-beta-methylbutyric acid in soybean peptide protein powder

A technology of methyl butyric acid and protein powder, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems that have not been reported before, and achieve the effects of economical effectiveness, good reproducibility, and high accuracy

Active Publication Date: 2017-04-26
GUANGZHOU HANFANG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, with the increasing attention to HMB in the above aspects, various food or preparation products containing HMB have appeared one after another, and there are no literatures on the detection of β-hydroxy-β-methylbutyric acid (HMB) content in these products. According to reports, β-hydroxy-β-methylbutyrate calcium is a new resource food approved by the Ministry of Health in 2011, and the enterprise standard is implemented

Method used

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  • Method for determining content of beta-hydroxy-beta-methylbutyric acid in soybean peptide protein powder
  • Method for determining content of beta-hydroxy-beta-methylbutyric acid in soybean peptide protein powder
  • Method for determining content of beta-hydroxy-beta-methylbutyric acid in soybean peptide protein powder

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Preparation of sample solution

[0023] Preparation of the sample solution: take 0.6g of the sample, accurately weigh it, put it in a 25ml measuring bottle, add 5ml of 0.1mol / L hydrochloric acid solution, shake vigorously to dissolve the solid, dilute to the mark with acetonitrile, shake for 1 minute, filter , Precisely draw 2.0ml of the filtrate, put it in a 5ml measuring bottle, dilute to the mark with 0.1mol / L hydrochloric acid solution, shake well, filter through a 0.45μm filter membrane, take the filtrate as the sample solution;

[0024] Preparation of negative solution: Take 0.6g of negative sample, weigh it accurately, put it in a 25ml measuring bottle, add 5ml of 0.1mol / L hydrochloric acid solution, shake vigorously to dissolve the solid, dilute to the mark with acetonitrile, shake for 1 minute, filter After that, accurately draw 2.0ml of the filtrate, put it in a 5ml measuring bottle, dilute to the mark with 0.1mol / L hydrochloric acid solution, shake well, ...

Embodiment 2

[0054] Embodiment 2: Optimization of sample dissolution process

[0055] Take 2.3g of the sample, put it in a 100ml measuring bottle, and directly dissolve the sample with 0.1mol / L hydrochloric acid solution. Remove protein from sample solution to eliminate interference.

[0056] Take 0.6g each of the sample and the negative sample, put them in a 25ml measuring bottle, add 5ml of 0.1mol / L hydrochloric acid solution, shake vigorously to dissolve the solid, dilute to the mark with acetonitrile, shake for 1 minute, filter, and accurately absorb the filtrate Put 2.0ml in a 5ml measuring bottle, dilute to the mark with 0.1mol / L hydrochloric acid solution, shake well, filter through a 0.45μm filter membrane, and take the subsequent filtrate as the sample solution. Organic solvents can reduce the electrolytic constant of the solution, increase the attraction of different charges on protein molecules, and reduce the solubility of proteins and precipitate them; therefore, acetonitrile...

Embodiment 3

[0057] Embodiment 3: the selection of mobile phase

[0058] (1) Prepare sample solution and negative sample solution as described in "Example 1";

[0059] (2) Preparation of the control solution: Weigh about 60 mg of the β-hydroxy-β-methylbutyrate calcium hydrate standard substance, put it in a 50ml measuring bottle, dissolve it with 0.1mol / L hydrochloric acid solution and dilute to the mark to obtain the control product solution.

[0060] (3) Chromatographic conditions:

[0061] Column: Phenomex Uranus C 18 Chromatography column, 250mm×4.6mm, 5μm.

[0062] Flow rate: 0.5ml / min; Column temperature: 35°C; Injection volume: 5μl;

[0063] Mobile phase 1: 0.01mol / L sodium heptanesulfonate solution-acetonitrile (95:5) (10% phosphoric acid solution to adjust the pH value to 3.0);

[0064] Mobile phase 2: 0.01mol / L potassium dihydrogen phosphate solution-acetonitrile (95:5) (10% phosphoric acid solution to adjust the pH value to 2.6);

[0065] Test with mobile phase 1 and mobil...

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Abstract

The invention discloses a method for determining the content of beta-hydroxy-beta-methylbutyric acid in soybean peptide protein powder. The method includes the steps that 1, during preparation of a sample solution, a sample is taken, 0.1 mol / L of a hydrochloric acid solution is added to dissolve the sample, then the mixture is diluted to a scale with acetonitrile, shaking and filtering, and subsequent filtrate is precisely sucked and diluted to a scale with 0.1 mol / L of a hydrochloric acid solution to serve as a sample solution after uniform shaking; 2, during preparation of a comparison production solution, beta-hydroxyl-beta-methyl butyrate hydrate is taken as a comparison product and diluted with 0.1 mol / L of a hydrochloric acid solution, and the comparison product is prepared, wherein the content of beta-hydroxyl-beta-methyl butyrate is 0.1 mg or 0.3 mg or 0.5 mg or 0.7 mg or 1.0 mg per ml of the comparison product; 3, during determining of the content of beta-hydroxyl-beta-methyl butyrate, a chromatographic column with octadecyl silane silica being a filling agent is adopted, 0.01 mol / L of sodium heptanesulfonate solution-acetonitrile serves as a mobile phase, the target component is eluted and separated, and the content of beta-hydroxyl-beta-methyl butyrate is calculated with a linear regression equation. The method can accurately and effectively determine the content of beta-hydroxyl-beta-methyl butyrate, and is simple and convenient to implement, economical, practical, good in repeatability and high in accuracy.

Description

technical field [0001] The invention belongs to the fields of formula food for special medical use, health food and medicine, and in particular relates to a method for determining the content of β-hydroxy-β-methylbutyric acid in soybean peptide protein powder by high performance liquid chromatography external standard method. Background technique [0002] β-Hydroxy-β-methylbutyric acid (HMB) is a five-carbon organic acid, which is a derivative of the essential amino acid leucine produced in vivo through its metabolite α-ketoisocaproic acid. A large number of studies have shown that HMB can prevent muscle loss and increase muscle mass, and is widely used in sports and bodybuilding. Many diseases, such as severe trauma, tumors, AIDS, and muscle atrophy in the elderly, can cause muscle loss. Therefore, in recent years, HMB has also been used in the clinical treatment of muscle wasting. At present, with the increasing attention to HMB in the above aspects, various food or prepa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/042
Inventor 王小妹温恺嘉李益罗明琍梁北梅曾荣华
Owner GUANGZHOU HANFANG PHARMA
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