Method and kit for high throughput testing of enzymatic activity in lipase in liquid milk product

A lipase enzyme and dairy product technology, which is applied in the field of high-throughput detection of lipase enzyme activity in liquid dairy products, can solve the problems of tedious time-consuming, large errors, etc., achieve less reagent usage, reduce product deterioration, and detect threshold low effect

Active Publication Date: 2016-06-08
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the lipase enzyme activity detection in the national standard adopts the sodium hydroxide titration method (QB / T1803-1993), using olive oil and polyvinyl alcohol emulsion as the reaction substrate, which belongs to a kind of titration method, which is cumbersome, time-consuming and expensive. big error

Method used

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  • Method and kit for high throughput testing of enzymatic activity in lipase in liquid milk product
  • Method and kit for high throughput testing of enzymatic activity in lipase in liquid milk product
  • Method and kit for high throughput testing of enzymatic activity in lipase in liquid milk product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1, UHT Skim Milk Lipase Enzyme Activity Detection

[0048] (1) Kit for detecting lipase enzyme activity

[0049] A kit for detecting biotinidase enzyme activity, comprising the following components:

[0050] Reagent 1:

[0051] Glycine-sodium hydroxide buffer solution, the pH value is 9.40, wherein the molar concentration of glycine is 150mM, and the molar concentration of sodium hydroxide is 56mM;

[0052] Reagent 2:

[0053] The acetonitrile solution of p-nitrophenyl palmitate, wherein the molar concentration of p-nitrophenyl palmitate is 50mM;

[0054] Reagent 3:

[0055] Stop solution, dairy clarifier.

[0056] (2) Drawing of lipase enzyme activity standard curve

[0057] 1) Preparation of UHT skim milk with different lipase activities

[0058] UHT skim milk with lipase activity of 0U, 0.01U, 0.05U, 0.10U, 0.2U, 0.3U and 0.4U were prepared respectively.

[0059] 2) Detection steps of lipase enzyme activity

[0060] a. Add 50 μl of Reagent 1 and 10 μl ...

Embodiment 2

[0074] Example 2, UHT whole milk lipase activity detection

[0075] (1) Kit for detecting lipase enzyme activity

[0076] Composition is the same as in Example 1.

[0077] (2) Standard curve of lipase enzyme activity

[0078] 1) Preparation of UHT whole milk with different lipase activity

[0079] UHT whole milk with lipase activity of 0U, 0.01U, 0.05U, 0.10U, 0.2U, 0.3U and 0.4U were prepared respectively.

[0080] 2) Detection steps of lipase enzyme activity

[0081] a. Add 50 μl of Reagent 1 and 10 μl of UHT whole milk with lipase activity of 0.01U, 0.05U, 0.10U, 0.2U, 0.3U and 0.4U into the wells of a 96-well microplate, shake and mix, and keep at 45°C Incubate for 5 minutes;

[0082] b. Add 10 μl of Reagent 2 to the wells of the microplate, shake and mix, and react at 45°C for 20 minutes;

[0083] c. Add 50 μl of Reagent 3 to the wells of the microplate, shake and mix, and place at 45°C for 5 minutes to terminate the reaction and clarify the system;

[0084] d. At ...

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Abstract

The invention discloses a method and kit for high throughput testing of enzymatic activity in lipase in a liquid milk product. The kit for testing of the enzymatic activity in the lipase in the liquid milk product comprises a reagent 1 and a reagent 2 which are independently packaged. The reagent 1 is a glycine-sodium hydroxide buffer solution, the pH value of the reagent 1 is 8.0-10.0, and the reagent 2 is an acetonitrile solution of p-nitrophenyl ester. The method is used for testing the enzymatic activity in the lipase in the liquid milk product through the kit, theoretical basis is provided for classification of raw milk, and fresh and raw milk quality control indexes suitable for producing and processing various kinds of liquid milk can be formulated, such as predicting UHT milk shelf life, scientifically guiding UHT product partition selling, lowering product metamorphism caused by residual lipase activity and protecting enterprise benefits and consumer benefits. The testing method is high in sensitivity, and the detection threshold value of lipase is low.

Description

technical field [0001] The invention relates to a method for high-throughput detection of lipase enzyme activity in liquid dairy products and a kit thereof, belonging to the field of food detection. Background technique [0002] At present, the consumption of liquid milk in our country is increasing year by year. It is an ideal nutritious food for human beings. However, due to microbial pollution and reproduction and the metabolites produced, the quality will deteriorate and seriously affect the health of the body. Raw milk is located at the most upstream of the dairy industry chain, and its quality will directly affect the quality and safety of dairy products. Raw milk is stored in the cold chain from milking to processing, and the contaminated microorganisms are mainly psychrophilic bacteria. During this period, some bacteria in the psychrophilic genus such as Pseudomonas fluorescens can develop heat resistance Even if the lipase is sterilized by instantaneous ultra-high ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 吕加平张维清逄晓阳刘鹭张书文芦晶
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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