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Method for detecting lipase activity inhibition rate

A technology of lipase and inhibition rate, applied in the field of detection, can solve the problems of unstable measurement results, reduction of fatty acid recovery rate, measurement error, etc.

Inactive Publication Date: 2012-05-23
BEIJING WBL PEKING UNIV BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct titration method is generally used for the determination of pure enzymes, and is not suitable for the determination of samples containing a large amount of impurities. The disadvantages of this method are the buffering effect of the buffer in the reaction system and the incomplete termination of the reaction, resulting in unstable determination results.
Extracting fatty acids with hydrophobic organic solvents and then titrating them is not only cumbersome to operate, but also for samples with complex components, if there is emulsification during the extraction process, the recovery rate of fatty acids will be reduced and measurement errors will also be caused.

Method used

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  • Method for detecting lipase activity inhibition rate
  • Method for detecting lipase activity inhibition rate
  • Method for detecting lipase activity inhibition rate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0211] A. Prepare reagents, olive oil emulsion and assay enzyme solution:

[0212] Tris buffer solution preparation: Take 3.030g of tris, adjust the pH value to 8.5 with 0.1mol / L hydrochloric acid solution, add water to 450-480ml, shake well, then adjust the pH value to 8.5, set Capacity up to 500ml;

[0213] Preparation of olive oil emulsion: Take 12ml of olive oil and 22.5g of gum arabic, grind them evenly, add water and grind to 300ml, and emulsify twice with a homogenizer at 18000 rpm, each time for 3 minutes; take the emulsion and examine it under a microscope, The diameter of 90% of the milk grains is less than 3 μm, and no milk grains of 10 μm are allowed;

[0214] Preparation of 8% ox bile salt solution: accurately weigh 8.0 g of ox bile salt, place in a 50 ml beaker, dissolve with water, transfer to a 100 ml volumetric flask to constant volume;

[0215] Preparation of 0.1mol / L sodium hydroxide solution: Accurately weigh 2.000g of sodium hydroxide, add water to disso...

Embodiment 2

[0227] A. Preparation of reagents, olive oil emulsion and determination of enzyme solution

[0228] Tris buffer solution preparation: Take 3.530g of tris, adjust the pH value to 7.8 with 0.1mol / L hydrochloric acid solution, add water to 450ml, shake well, then adjust the pH value to 7.8, and dilute to 450ml;

[0229] Preparation of olive oil emulsion: Take 14ml of olive oil and 24g of gum arabic, grind them evenly, add water and grind to 350ml, emulsify twice with a homogenizer at 18000 rpm, each time for 3 minutes; take the emulsion and check it under a microscope, 90 % The diameter of the milk grains is less than 3 μm, and no milk grains of 10 μm are allowed;

[0230] Preparation of 6% ox bile salt solution: accurately weigh 6.0 g of ox bile salt, place in a 50 ml beaker, dissolve with water, transfer to a 100 ml volumetric flask to constant volume;

[0231] Preparation of 0.1mol / L sodium hydroxide solution: Accurately weigh 2.000g of sodium hydroxide, add water to dissolv...

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Abstract

The invention discloses a method used for measuring suppression ratio of the activity of lipase. The method comprises the steps: A. reagents and olive oil emulsified mixture are prepared, and enzyme liquid is measured; B. an activity measuring system of the lipase is set up; C. the specification curve of the relation between the suppression ratio 'Y%' of the lipase and pH range of variation 'delta pH'; D. the suppression ratio of the lipase of sample solution is measured. The measuring method takes lipase activity as target spot, and uses containment procedure of the lipase activity as index; according to the action principle of the lipase, a method used for measuring the suppression ratio of the lipase. The measuring method is simple, convenient and reliable, has definite target spot and clear mechanism, can be used for screening various materials having the inhibitory action on the lipase in a large quantity, and can be further applied for screening and developing medicine for losing weight and decreasing blood lipids.

Description

technical field [0001] The invention relates to a detection method, in particular to a method for detection of lipase activity inhibition rate. Background technique [0002] There are many methods for the determination of lipase activity, including acid-base titration, potentiometric titration, turbidimetry, and spectrophotometry. However, the above method is cumbersome to operate, and the detection error is relatively large due to the influence of the purity of the detection sample. At present, there is no uniform detection method for lipase activity. Manufacturers and scientific research departments usually formulate their own product detection methods according to their own experimental conditions and actual conditions and the characteristics of the lipase they prepare. For scientific research and industrial production using lipase, its activity is a very important indicator, so it is very important to determine the accuracy and comparability of activity detection. [0...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/34
Inventor 段震文郭树仁何大林闫雪秋
Owner BEIJING WBL PEKING UNIV BIOTECH