Method for detecting 18 varieties of protein hydrolytic amino acids in milk powder through high-performance liquid chromatographic method

A high-performance liquid chromatography and proteolysis technology, which is applied in the field of high-performance liquid chromatography detection of 18 kinds of proteolytic amino acids in milk powder, can solve the problems of poor universality and high cost, and achieve the effect of strong universality

Inactive Publication Date: 2012-01-18
山西出入境检验检疫局检验检疫技术中心
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Problems solved by technology

[0007] The present invention provides milk powder in order to solve the problems of high cost and poor universality of the existing amino acid determination method and the high-performance liquid chromatography-based amino acid determination method cannot meet the requirements of fast and accurate qualitative and quantitative analysis of 18 kinds of proteolytic standard amino acids. High performance liquid chromatography detection method for 18 kinds of proteolytic amino acids in

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  • Method for detecting 18 varieties of protein hydrolytic amino acids in milk powder through high-performance liquid chromatographic method
  • Method for detecting 18 varieties of protein hydrolytic amino acids in milk powder through high-performance liquid chromatographic method
  • Method for detecting 18 varieties of protein hydrolytic amino acids in milk powder through high-performance liquid chromatographic method

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[0032] The present invention will be further described below in combination with the accompanying drawings and specific embodiments.

[0033] The high-performance liquid chromatography detection method of 18 kinds of proteolytic amino acids in milk powder comprises the following steps:

[0034] (1) Preparation of milk powder hydrolysis samples:

[0035] Three different pretreatment methods, i.e. conventional hydrolysis, oxidative hydrolysis, and alkali hydrolysis, were used to hydrolyze the milk powder to obtain milk powder hydrolyzed samples respectively; the milk powder was selected as infant growth milk powder, and the following scheme could be adopted for specific implementation:

[0036]① Conventional hydrolysis method: Weigh 0.0500g (accurate to 0.0001g) of milk powder and transfer it to an ampoule or hydrolysis tube, add 10ml of 6mol / L hydrochloric acid, add 2-3 drops of dissolved fresh phenol, and fill it with high-purity nitrogen for 15 minutes , Seal the ampoule bot...

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Abstract

The invention belongs to the technical field of amino acid detection, in particular relates to a method for detecting 18 varieties of protein hydrolytic amino acids in milk powder through a high-performance liquid chromatographic method, solving the problems that the traditional amino acid determination method has high cost and poor universality and the qualitative and quantitative analysis of the 18 varieties of standard amino acids after proteolysis cannot be quickly and accurately performed through a high-performance liquid chromatograph at present. The method comprises the steps of: pre-treating the milk powder through three different hydrolysis methods; performing pre-column derivatization of a hydrolyzed milk powder smaple through a CNBF (4-chloro-3,5-dinitrobenzotrifluoride) derivatizing agent; optimizing derivatization conditions and chromatographic fractionation detection conditions by adopting the HPLC (High-performance Liquid Chromatography) and a universal C18 (250mm*4.6mm, 5mu m) liquid chromatography column; and separating and determining all 18 varieties of proteolyzed amino acids in the milk powder. The method has the advantages of stronger universality and no need of the precious amino acid analyzer and/or the precious amino acid determination reagent kit, thereby being suitable for more common laboratories.

Description

technical field [0001] The invention belongs to the technical field of amino acid detection, in particular to a high performance liquid chromatography detection method for 18 proteolytic amino acids in milk powder. Background technique [0002] In recent years, dairy food safety incidents have aroused widespread concern in the society. Due to the obvious increase in foodborne diseases caused by dairy products, the quality and safety of dairy products has received extensive attention. While the development of food science and technology has enriched human food consumption, it has also brought safety issues to dairy production. In order to strengthen the quality supervision of dairy products, "Food Safety Law", "Regulations on the Supervision and Administration of Dairy Product Quality and Safety", "2010 Detailed Rules for the Examination of the License Conditions for the Production of Dairy Products by Enterprises", "2010 Version of the Detailed Rules for the Review of th...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/60
Inventor 李卫华傅英文宋欢李玉玲张建军
Owner 山西出入境检验检疫局检验检疫技术中心
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