High-performance liquid chromatography method for measuring nucleotide in milk powder

A technology of high performance liquid chromatography and determination method, which is applied in the field of high performance liquid chromatography determination of nucleotides in milk powder. The effect of high accuracy, complete separation and simple method

Active Publication Date: 2012-09-19
SHANDONG KAISHENG NEW MATERIALS
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  • Abstract
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Problems solved by technology

According to related data, when processing samples, it is generally used to add a certain amount of acetic acid or perchloric acid to the sample solution to precipitate the protein, and then separate and remove it. This method has been proved by many tests to be unsatisfactory.
Add acetic acid or perchloric acid to the sample solution to precipitate protein, but the protein precipitate is vi

Method used

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  • High-performance liquid chromatography method for measuring nucleotide in milk powder

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Embodiment 1

[0026] Nucleotide content detection in milk powder:

[0027] 1. Preparation of nucleotide standard reference solution:

[0028] 1.1 Preparation of reference substance stock solution: Weigh 200mg (accurate to 0.1mg) in a 50ml volumetric flask, dissolved in distilled water and made to volume.

[0029] 1.2 Preparation of the reference substance solution: draw 2.00ml of the reference substance stock solution into a 25ml volumetric flask, and distill to volume with distilled water.

[0030] 2. Sample dissolution:

[0031] Weigh 2.00 g of commercially available milk powder samples containing 5 kinds of nucleotides (cytidylic acid, disodium uridylate, disodium guanylate, adenylic acid, disodium inosinate) into a 100ml beaker, add 30ml of distilled water at 64~66℃, shake well, and sonicate in an ultrasonic machine for 10 minutes.

[0032] 3. Separation of sample protein:

[0033] According to the pH meter, adjust the pH value to 3.5~3.8 with nitric acid solution (1+4) to make the...

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Abstract

The invention relates to a high-performance liquid chromatography method for measuring nucleotide in milk powder, and belongs to the technical field of detection for food additives. The method comprises the following steps of: preparing a contrast solution by using a nucleotide standard substance; weighing a test sample, dissolving the test sample in distilled water through ultrasonic oscillation, so that the nucleotide in the milk powder is dissolved in the distilled water; adjusting the pH value of the test sample solution by hydrogen nitrate until the pH value is 3 to 4, precipitating proteins, filtering and removing the proteins, fixing the volume of a filtrate by using water to obtain a to-be-tested solution; measuring nucleotide chromatographic peak areas in the test solution and the contrast solution by high performance liquid chromatography; and calculating the nucleotide content in the test sample by an external standard method according to the concentration of the nucleotidein the contrast solution, the nucleotide chromatographic peak area in the test solution and the mass of the test sample. The method is simple, convenient to operate and high in precision; proteins can be completely separated, so that interference with the measurement for the nucleotide is eliminated; and a good condition is supplied to measurement of the nucleotide in the test sample.

Description

technical field [0001] The invention relates to a high-performance liquid chromatography determination method for nucleotides in milk powder, which belongs to the technical field of food additive detection. Background technique [0002] Nucleotide is a nutrient that plays an important role in the growth and development of infants, so nucleotides can be used in dairy additives. However, the human body's demand for nucleotides is limited, and the more the better, the content of nucleotides in dairy products must be controlled. Nucleotide content is listed as an important indicator in the milk powder standards of dairy product manufacturers, so it is extremely important to control the content of nucleotides in dairy products, which requires corresponding technical support. The currently reported detection methods mainly include microbiological method, capillary electrophoresis method, high performance liquid chromatography and so on. Among them, high-performance liquid chroma...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/14
Inventor 张振霞及汉俊孙丰春李雪艳李文娟李云龙毕义霞
Owner SHANDONG KAISHENG NEW MATERIALS
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