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ELISA detection method of dioscin content

A technology of diosgenin and detection method, which is applied in the direction of measuring device, biological test, material inspection product, etc., can solve the problems of difficulty in simultaneous detection of a large number of samples, complicated operation, expensive equipment, etc., and achieves continuous and rapid detection, high sensitivity, and detection. The effect of process simplification

Inactive Publication Date: 2004-12-15
WUHAN UNIV
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AI Technical Summary

Problems solved by technology

[0007] Wherein gravimetric method and colorimetric method require simple equipment, disadvantage is time-consuming, it is difficult to carry out the simultaneous detection of a large number of samples, and the error is relatively large; High performance liquid chromatography is accurate in measuring result, but also time-consuming, and the required equipment is expensive (market It costs hundreds of thousands to millions of yuan to sell a liquid chromatograph), the operation is complicated, and operators who need special training are beyond the reach of ordinary enterprises or individuals.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0041] Sample Preparation:

[0042] Weigh an appropriate amount of pulverized sample to be tested (0.01g-1g, depending on the sample), add methanol for ultrasonic extraction (if there is no ultrasonic equipment, just let it stand overnight) 3 times, 3-5ml each time, and combine the extraction solution , constant volume, activated carbon decolorization.

[0043] Content detection:

[0044] Coat the ELISA plate with the coating material (overnight at 37°C), wash with phosphate buffer saline → add standard diosgenin with gradient concentration → add rabbit anti-diosgenin and related saponin antibody (incubate at 37°C for 1 hour), phosphate buffer saline Washing → add enzyme-labeled goat anti-rabbit (incubate at 37°C for 1 h), wash with phosphate buffer solution → add substrate and develop color at room temperature in the dark for about 15 min → measure OD value on a microplate reader and draw a standard curve.

[0045] Take the sample to be tested, dilute it appropriately, dete...

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PUM

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Abstract

The present invention relates to the method of detecting dioscin content in plant body or relevant sample, and is ELISA detection method. Protein link coupled dioscin is used as antigen to obtain dioscin and its relevant saponin antibody, and the dioscin and its relevant saponin antibody is used as solid antigen in competitive enzynme linked immunological detection of the dioscin content in plant body or relevant sample. The method is simple, low in cost and high in sensitivity, and may be used in different departments.

Description

technical field [0001] The invention relates to a method for detecting the content of diosgenin in plants, in particular to a method for quantitatively detecting the content of diosgenin in plants by using ELISA (enzyme-linked immunosorbent assay). Background technique [0002] Diosgenin is the most important raw material for the synthesis of steroid hormone drugs. Due to the cumbersome and time-consuming steps of conventional methods for detecting diosgenin content in plants, it is an urgent problem to be solved to improve the determination method of diosgenin content in plants. [0003] As far as is known, the conventional methods for detecting the content of diosgenin in plants (usually yam plants) include: [0004] ① Gravimetric method: Weigh two parts of the sample to be tested, chopped and dried in the sun (or dry at 80°C, and measure the moisture separately), each 30g, add 300ml of 2N hydrochloric acid respectively, heat and hydrolyze in a water bath for 3.5 hours, s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N33/535G01N33/543
Inventor 李家儒何骥胡江丽
Owner WUHAN UNIV
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