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Method for detecting yeast beta-glucan in milk or milk product

A detection method and technology for dairy products, applied in the field of analysis and detection, can solve problems such as the lack of detection standards and related technologies, affecting the extraction and purification of yeast β-glucan, and the complexity of the dairy matrix

Active Publication Date: 2016-07-06
COFCO GROUP +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the dairy matrix is ​​extremely complex, and the fat globules, protein bundles, stabilizers and flavor ingredients in it will affect the extraction and purification of yeast β-glucan, so how to accurately determine the yeast β-glucan in milk and dairy products Sugar remains a challenging subject
At present, only the AOAC32.2.10 standard "AOAC Official Method 995.16β-D-GlucaninBarleyandOats" and the agricultural industry standard NY / T2006-2011 "Determination of β-glucan content in grains and their products" detect water-soluble β-glucan Standardization has been carried out, but for water-insoluble β-glucan, especially yeast β-glucan in milk and dairy products, there is a lack of mature detection standards and related technologies at home and abroad

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  • Method for detecting yeast beta-glucan in milk or milk product
  • Method for detecting yeast beta-glucan in milk or milk product
  • Method for detecting yeast beta-glucan in milk or milk product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: the relation of the dosage of neutral protease and alkaline protease, enzymolysis time and sample proteolysis degree

[0052] In this example, the degree of proteolysis is used as an index to measure the degree of proteolysis.

[0053] 0.02%, 0.05%, 0.1%, 0.15% and 0.2% (v / v) by volume to milk supplemented with yeast β-glucan neutral protease stock solution and Equal-volume mixed solution of alkaline protease stock solution (converted according to the density mentioned above, in the milk to be enzymatically hydrolyzed, the enzyme activity of Neutrase neutral protease is 1.0×10 -4 AU / mL, 2.5×10 -4 AU / mL, 5.0×10 -4 AU / mL, 7.5×10 -4 AU / mL, 1.0×10 -3 AU / mL, the enzyme activity of Alcalase alkaline protease is 2.8×10 -4 AU / mL, 7.0×10 -4 AU / mL, 1.4×10 -3 AU / mL, 2.1×10 -3 AU / mL, 2.8×10 -3 AU / mL), the enzymatic hydrolysis was carried out at 45°C for 6 hours. After the enzymatic hydrolysis was completed, the degree of proteolysis of each sample was det...

Embodiment 2

[0055] Embodiment 2: the mensuration of actual sample

[0056] Samples 1-12 (see Table 1 for sample numbers) are all milk samples added with 42 mg / 100 g of yeast β-glucan.

[0057] 1) Pretreatment step: add 0.1% (v / v) to 12 yeast β-glucan milk samples respectively neutral protease stock solution and Equal volumes of the alkaline protease stock solution were hydrolyzed at 45° C. for 2 hours, and the obtained hydrolyzed product was centrifuged at 10,000 rpm. The supernatant was discarded, and the obtained crude dextran precipitate was washed twice with pure water. Then add 1 mol / L hydrochloric acid to the precipitate and hydrolyze at 121°C for 1 hour.

[0058] 2) Analysis step: the glucose in the sample after the pretreatment step is determined by ion chromatography. Before the ion chromatographic analysis, the dextran hydrolysis solution obtained from the pretreatment was diluted by direct dilution method, so that the concentration of glucose to be determined was in the l...

Embodiment 3

[0076] Embodiment 3: Sensitivity, accuracy and precision analysis of the method

[0077] Use pure water to prepare glucose standard solutions and sample solutions with a series concentration of 0.5, 1.0, 4.0, 8.0, 10.0, 15.0, and 20.0 μg / mL, and evaluate the sensitivity, accuracy, and precision of the analysis.

[0078] Under the same chromatographic conditions as in Example 2, the linear regression equation obtained by measuring the glucose standard solution is Y=2.865X+0.395, the linear range is 0.5~20 μg / mL, the linear correlation coefficient is 0.9995, and the detection limit is 0.1 μg / mL. The relative standard deviation was 0.15% (n=6).

[0079] Taking the concentration of yeast β-glucan added in dairy products at 42.00mg / 100g as the standard sample, prepare samples with three addition levels of 70%, 100% and 130% of the standard sample, and determine the sample recovery rate at the three concentrations : when the addition amount was 70%, the measured value was 22.32mg / 1...

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Abstract

The invention relates to a method for detecting yeast beta-glucan in milk or a milk product. The method comprises 1, pretreatment: carrying out enzymolysis on milk or a milk product through a neutral protease or an alkaline protease, carrying out centrifugation on the enzymolysis product, cleaning the precipitates obtained through centrifugation through pure water or deionized water, adding an monobasic acid into the precipitates and carrying out heating hydrolysis, and 2, analysis: determining glucose content of the pretreatment product through ion chromatography and calculating yeast beta-glucan content of the milk or milk product.

Description

technical field [0001] The invention relates to an analysis and detection method, in particular to a detection method of yeast beta-glucan in milk or milk products. Background technique [0002] Yeast β-glucan (yeast betaglucan) is a glucose polymer extracted from the cell wall of yeast. Its molecular structure main chain contains β-1,3-D glycosidic bonds, while the branch chain contains β-1,6-D glycosides key, the ratio of the two is about 85:15. The linear molecular polymerization degree of yeast β-glucan is 1500, the molecular weight is about 240kDa, and the branched molecular polymerization degree is 140, the molecular weight is about 22kDa. As an important source of food dietary fiber, yeast β-glucan has many nutritional values ​​and physiological functions such as enhancing immunity, preventing oxidative radiation, anti-tumor inflammation, reducing blood lipid cholesterol, and promoting wound healing. In June 2010, Ministry of Health Announcement No. 9 approved yeast...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 杨永坛谢云峰任丹丹刘佟杨悠悠
Owner COFCO GROUP
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