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Method for measuring nucleotide content in dairy product

A determination method and nucleotide technology, which are applied in the field of determination of nucleotide content in dairy products, can solve problems such as the lack of suitable methods for samples, and achieve the effects of being easy to master and easy to operate.

Inactive Publication Date: 2015-03-04
西安莹朴生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The detection methods currently reported mainly include microbiological method, ultraviolet spectrophotometry, high performance liquid chromatography, etc., but most of them are developed for nucleotide additives, and there is no suitable method for samples with complex components such as dairy products. method

Method used

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  • Method for measuring nucleotide content in dairy product
  • Method for measuring nucleotide content in dairy product

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Experimental program
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Effect test

Embodiment 1

[0018] (1) Preparation of standard solution: Prepare GMP, UMP, GMP, IMP, and AMP stock solutions with a concentration of 1 mg / ml; then prepare a series of standard concentrations with concentrations of 10.0, 20.0, 30.0, 40.0, and 50.0 μg / ml The solution, the injection volume is 10μl respectively, through the high performance liquid chromatography on Dima C18 column, 5μm 4.6*200mm, column temperature 25℃, mobile phase flow rate 1.0ml / min, UV detector wavelength 254nm, mobile phase is Under the conditions of the mixture of 0.01mol / L potassium dihydrogen phosphate 960ml+0.1mol / L dipotassium hydrogen phosphate 40mL, the relationship curve and correlation coefficient between peak height and concentration were measured.

[0019] Thus, the relationship curve of peak height-concentration is shown in Table-1.

[0020] Table-1 Standard Curve

[0021]

[0022] According to table-1, the regression equation of five kinds of nucleotide standard curves can be calculated as:

[0023] CMP...

Embodiment 2

[0036] Embodiment two: recovery rate and reproducibility test

[0037] (1) The determination method of the present invention was repeated 6 times, and the recovery rate was between 92% and 97%, with an average recovery rate of 94.6%, and its accuracy could meet the requirements for determination of infant formula milk powder.

[0038] (2) assay method of the present invention carries out 8 parallel sample assays, and result statistics shows, the repeatability of assay result is better, coefficient of variation 1.79%, can satisfy adopting and measure the requirement of this glycoside content in infant formula milk powder.

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Abstract

The invention discloses a method for measuring nucleotide content in a dairy product and belongs to the field of food additives and detection methods thereof. The method comprises the following steps: (1), preparing stock solutions of GMP, UMP, CMP, IMP, AMP, wherein the concentrations of the stock solutions are 1 mg / ml, and preparing into solutions with the series normal concentrations of 10.0 [mu]g / ml, 20.0 [mu]g / ml, 30.0 [mu]g / ml, 40.0 [mu]g / ml and 50.0 [mu]g / ml, wherein the sample size is 10 [mu]l, measuring the peak heights of the solutions through a liquid chromatograph and calculating a regression equation; (2), weighing 10.0 g of a sample, dissolving the sample with 20 ml of warm water, putting into an ultrasonic bath for oscillation for 10 min, then taking out, transferring into a 50-ml volumetric flask, adding acetic acid for protein precipitation, after diluting, uniformly mixing, filtering through filtering paper, filtering through a filtering membrane, and measuring the peak height of the sample through the high performance liquid chromatograph; (3), according to a standard curve equation and the peak height value of the sample, obtaining the nucleotide content in the dairy product. According to the invention, the liquid chromatography is adopted to measure the nucleotide content in an infant formula milk powder, the method is simple and convenient to operate, the measurement precision and accuracy can meet analysis requirements of the dairy product, and the method can be used as a normal inspection method.

Description

technical field [0001] The invention relates to a method for measuring nucleotide content in dairy products, and belongs to the field of food additives and detection methods thereof. Background technique [0002] Nucleotides are the building blocks of nucleic acids and are currently used in dairy products primarily in infant formula. There are five main kinds of nucleotides added as additives --- cytosine nucleoside (CMP), uridine nucleoside (UMP), adenosine nucleoside (AMP), guanosine nucleoside (GMP), hypoxanthine nucleus Glycosides (IMPs). In addition, it can also be added to beverages, baked goods, and various main and non-staple foods and beverages. The main physiological function of nucleotides is to widely participate in the energy metabolism and protein synthesis of the human body, and to increase the activity of various enzymes, activate cell functions, promote antibody formation, and enhance immune function. They have a good health care effect on the human body,...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 李勇刘文斌
Owner 西安莹朴生物科技股份有限公司
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