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Method for detecting activity of lipase in raw milk and application of method

A detection method, lipase technology, applied in the direction of measuring devices, analysis materials, color/spectral characteristics measurement, etc., can solve the problems of imperfect detection methods, poor accuracy, and long time consumption, and achieve fast detection speed, accurate and reliable results, Simple effect of reagent type

Active Publication Date: 2021-09-24
四川新华西乳业有限公司
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  • Description
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AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is: the purpose of the present invention is to provide a detection method of lipase activity

Method used

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  • Method for detecting activity of lipase in raw milk and application of method
  • Method for detecting activity of lipase in raw milk and application of method
  • Method for detecting activity of lipase in raw milk and application of method

Examples

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Embodiment 1

[0098] Preparation of lipase dilution

[0099]Accurately weigh 0.025g 20000U / g lipase powder and dissolve it in 100mL UP water, mix well to make 5U / mL lipase mother solution; pipette 1ml 5U / ml lipase mother solution into 9ml ultrapure water, mix well Obtain 500mU / mL secondary mother liquor of lipase, pipette 1mL of secondary mother liquor into 4mL ultrapure water, obtain 100mU / mL lipase dilution, set aside.

[0100] The lipase mother liquor is stored in a brown frontal bottle, protected from light and kept refrigerated at 2-6°C, and the validity period is 3 days. The diluted lipase solution should be prepared immediately.

[0101] Preparation of phosphate buffer:

[0102] Measure 200mM 14mL sodium dihydrogen phosphate solution and 200mM 36mL disodium hydrogen phosphate solution, add 150mL UP water, add 0.1mL antibiotic solution, mix well to get 50mM phosphate buffer solution, pH=7.2, 2-6℃ Keep refrigerated and the validity period is 2 months. Wherein sodium dihydrogen phos...

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Abstract

The invention relates to the technical field of food analysis and detection and particularly relates to a method for detecting the activity of lipase in raw milk and application of the method. The method comprises the following steps of preparing a sample diluent and a lipase diluent; then adding a phosphate buffer solution and a substrate solution to obtain a sample to be detected and a labeled sample; 4-nitrophenyl octanoate being used as a substrate, lipase powder being used for preparing lipase diluents with different gradients to decompose the substrate, a substance capable of absorbing the wavelength of 316 nm being generated, so an absorbance-lipase activity function relation being established; and then carrying out centrifugal treatment on the raw milk, diluting the lower clear liquid, testing the absorbance of the to-be-tested sample and the labeled sample by using an ultraviolet spectrophotometer, and judging activity of the lipase in the raw milk according to the absorbance-lipase activity function relationship.

Description

technical field [0001] The invention relates to the technical field of food analysis and detection, in particular to a detection method and application of lipase activity in raw milk. Background technique [0002] There are two main sources of lipase in milk, one is naturally present in raw milk, called natural lipase, and the other is produced by microbial metabolism in milk, called microbial lipase. The microbial lipase in milk is mainly derived from psychrophilic bacteria, although the reproduction of psychrophilic bacteria itself will not seriously affect the stability of milk, and it is easy to kill them in relatively mild heat treatment (such as pasteurization) , but the metabolites of some psychrophilic bacteria are heat-resistant lipases, and these heat-resistant lipases cannot be completely inactivated after pasteurization or even ultra-high temperature sterilization. Residual heat-resistant lipase will decompose the fat globular membrane and fat in milk, produce f...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/01
CPCG01N21/31G01N21/01G01N2021/0112
Inventor 陈柏锡王琴曾燕萍黄艳艳何刚沙马志喜李斌胡燕群
Owner 四川新华西乳业有限公司
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