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Method for measuring content of amino acid in black-bone chicken milk increasing preparation

A determination method and amino acid technology, which is applied in the field of determination of amino acid content in black-bone chicken milk-enhancing preparations, can solve the problems of complex pretreatment, low specificity, and difficult operation

Inactive Publication Date: 2016-01-06
天津飞鹰玉川药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the determination method of amino acids, the current quality standard is the State Food and Drug Administration standard WS 3 -461(Z-054)-2005(Z), in which the determination of protein content is only the determination method of total nitrogen, this method can only explain the content of nitrogen in the product, but cannot explain the content of protein, especially the content of active ingredients of amino acids
In recent years, there are mainly spectrophotometry, high performance liquid chromatography (HPLC) method, ion chromatography, liquid chromatography-mass spectrometry, and amino acid analyzer method for amino acid analysis and determination at home and abroad; spectrophotometry can only solve the problem of the total amount of amino acids. Analysis, unable to determine the type of amino acid; the pretreatment of HPLC and ion chromatography is complex and needs to be derivatized before determination, and the specificity is not strong; the instrument of liquid mass spectrometry is expensive, the pretreatment requirements are high, and the operation is difficult

Method used

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  • Method for measuring content of amino acid in black-bone chicken milk increasing preparation
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  • Method for measuring content of amino acid in black-bone chicken milk increasing preparation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The preparation of embodiment 1 amino acid mixed standard solution

[0050] Take an appropriate amount of each of the 17 kinds of amino acid standard products, weigh them accurately, put them in the same 250ml measuring bottle, add 0.02mol / L hydrochloric acid solution to dissolve and dilute to the mark, constant volume, shake well, and use it as the standard product stock solution. Then accurately measure 5ml of the standard stock solution and place it in a 50ml measuring bottle, add 0.02mol / L hydrochloric acid solution to dilute to the mark, constant volume, shake well, and use it as the standard solution. The sample volume and concentration of each amino acid standard are shown in Table 3.

[0051] Table 3 The sample volume and concentration table of each amino acid standard

[0052]

[0053]

[0054] Note: The concentration of lysine hydrochloride in standard stock solution and standard solution has been converted into lysine.

Embodiment 2

[0055] The preparation of embodiment 2 test solution

[0056] Precisely weigh about 55mg of black-bone chicken milk-enhancing preparation and place it in a hydrolysis tube, add 10ml of 6mol / L hydrochloric acid solution, fill it with nitrogen and seal it, place the sealed hydrolysis tube in a constant temperature drying oven at 110°C, hydrolyze it for 22 hours, take it out and cool it. Open the hydrolysis tube, transfer all the hydrolyzate to an evaporating dish, place it on a water bath and evaporate to dryness, add an appropriate amount of 0.02mol / L hydrochloric acid solution to the residue to dissolve and transfer it to a 50ml measuring bottle, add 0.02mol / L hydrochloric acid solution to dilute to scale, constant volume, shake well, filter with a microporous membrane, and take the filtrate as the solution to be tested.

Embodiment 3

[0057] Embodiment 3 chromatographic determination

[0058] 20 μl each of the standard solution and the solution to be tested (batch number 1307008) were accurately drawn, respectively, and injected into the amino acid analyzer for determination.

[0059] Wherein, proteolysis analysis buffer solution B1: PH-1; B2: PH-2; B3: PH-3; B4: PH-4; B5: water; B6: PH-RG. Reaction solution R1: ninhydrin; R2: ninhydrin buffer; R3: 5% ethanol. The analysis method was carried out according to the procedures in Table 4.

[0060] Table 4 Analysis method program table

[0061]

[0062]

[0063] Detection wavelength: channel 1: 570nm; channel 2: 440nm (proline detection); chromatographic column temperature: 57°C; reaction column temperature: 135°C; injection volume: 20μl.

[0064] The retention time and resolution results of each amino acid are shown in Table 3.

[0065] Table 5 Retention time and resolution of each amino acid

[0066]

[0067]

[0068] It can be seen from Table...

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Abstract

The invention discloses a method for measuring the content of amino acid in a black-bone chicken milk increasing preparation. The method comprises the following steps: subjecting a black-bone chicken milk increasing preparation to a protein hydrolysis treatment; and then using an amino acid analyzer to analyze the hydrolysate so as to measure the amino acid content. In the conventional standards, the protein content can be only represented by total nitrogen content, the provided method can overcome the shortage, moreover, the method can precisely measure contents of 17 amino acids in the preparation, has the advantages of simple pretreatment, strong specificity, good repeatability, and high precision, can well control the quality of the black-bone chicken milk increasing preparation, and improves the safety and effectiveness of the preparation.

Description

technical field [0001] The invention relates to an application method of quality control in the field of pharmacy, in particular to a method for simultaneously determining the content of 17 kinds of amino acids in a black-bone chicken milk-enhancing preparation. Background technique [0002] The black-bone chicken milk-enhancing preparation is a good medicine for nourishing qi and blood, making qi and blood biochemical and promoting milk secretion, and amino acid is an active ingredient of tonic medicine. For the determination method of amino acids, the current quality standard is the State Food and Drug Administration standard WS 3 -461(Z-054)-2005(Z), in which the determination of protein content is only a method of measuring total nitrogen, which can only explain the content of nitrogen in the product, but cannot explain the content of protein, especially the content of active ingredients of amino acids. In recent years, there are mainly spectrophotometry, high performan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 李凤
Owner 天津飞鹰玉川药业有限公司
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