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Method for detecting activity of hepatocyte auxin

A technology for detecting hepatocyte growth factor and activity, applied in the field of detection, can solve the problems of unstable test, low stimulation index value, poor consistency, etc., and achieve the effects of good measurement consistency, reliable measurement results, and stable measurement methods.

Active Publication Date: 2008-07-23
HANGZHOU HUAJIN PHARMA
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Problems solved by technology

During the detection, it was found that due to the glutamine component in the RPMI 1640 medium of the prior art detection method, glutamine can promote the growth of liver cancer cells during the activity detection process, resulting in a relatively high absorption value of the detected cell control group, which in turn led to the detection calculation The stimulation index value obtained is lower than the actual stimulation index value of the test product, which makes the test unstable and the consistency of repeated measurements is poor, so that it is impossible to correctly judge the activity of the test product hepatocyte growth-stimulating hormone.

Method used

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  • Method for detecting activity of hepatocyte auxin
  • Method for detecting activity of hepatocyte auxin

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Embodiment Construction

[0008] Below by embodiment, the present invention is described further.

[0009] The hepatocyte growth-promoting hormone activity detection method uses RPMI-1640 culture fluid without glutamine components. The RPMI-1640 culture fluid without glutamine components can use finished reagents, and can also use glutamine-free RPMI-1640 culture fluid. RPMI-1640 medium preparation.

[0010] Embodiment Hepatocyte growth-promoting hormone activity detection method divides reagent preparation, needs testing product solution preparation, measures need testing product group, control group and blank control group each 3 hole absorbance average value, calculates stimulation index four steps:

[0011] 1. Preparation of reagents such as 0.01mol / L phosphate buffer solution, RPMI-1640 culture solution, 10% calf serum culture solution, 0.25% trypsin-edetate disodium digestion solution and MTT thiazole blue solution:

[0012] ① Prepare 0.01mol / L phosphate buffer solution with pH 7.3, take 8.0g of...

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Abstract

The invention discloses an activity check method of hepatocyte growth-promoting factor, which uses RPMI-1640 culture solution. The inventive method comprises agent preparation step, sample solution preparation, sample group 3-pore average absorbance test, control group 3-pore average absorbance test and blank control group 3-pore average absorbance test and excite index calculation. The invention is characterized in that the RPMI-1640 culture solution is the one without glutamine. The invention overcomes the defects of prior art as instable test and low repeated measurement consistency, with stable test method, reliable test result, high test data repetition rate and better consistency, while the test result can correctly present sample activity.

Description

technical field [0001] The invention relates to a detection method, in particular to a detection method for hepatocyte growth-promoting hormone activity. Background technique [0002] Hepatocyte growth-stimulating hormone is a biochemical drug extracted from fresh suckling pig liver or calf liver. As an auxiliary drug for the treatment of various severe hepatitis, chronic active hepatitis and liver cirrhosis, its activity detection is very important . The national drug standard WS-10001-(HD-0860)-2002 of the Drug Administration of the People's Republic of China stipulates that the hepatocyte growth-promoting hormone activity is determined by the MTT method listed in Attachment 1. The method uses RPMI 1640 medium containing glutamine components, including reagent preparation, preparation of the test solution, determination of the average absorbance of the 3 wells of the test product group, the average absorbance of the 3 wells of the control group and the 3 wells of the blan...

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Application Information

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IPC IPC(8): G01N21/00C12Q1/02
Inventor 周正兵俞保彬高留根
Owner HANGZHOU HUAJIN PHARMA
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