Method applying HPLC and simultaneously determining phytoecdysone substance

A high-performance liquid chromatography, phytoecdysone technology, which is applied in the field of analytical chemistry, can solve the problem of not being able to detect multiple ecdysone substances at the same time, and achieves the effects of rapid separation, high sensitivity and easy operation.

Inactive Publication Date: 2014-06-04
QINGDAO AGRI UNIV
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Problems solved by technology

[0004] However, none of the existing detection methods at home and abroad can simultaneously detect the above-mentioned multiple ecdysone substances under the same chromatographic conditions.
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  • Method applying HPLC and simultaneously determining phytoecdysone substance

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[0022] The technical solutions of the present invention will be further described below in conjunction with the drawings and specific embodiments.

[0023] A high-performance liquid chromatography method for simultaneous determination of phytoecdysone substances includes the following steps:

[0024] 1. Use YMC250×4.6mm, S-5μm chromatographic column;

[0025] 2. Use methanol-water mobile phase for gradient elution. The gradient elution procedure is 0-30min, methanol 30%→95%; 30-40min, methanol 95%→100%; 40-60min, methanol 100%; 60 -65min, methanol 100%→30%; 65-70min, methanol 30%;

[0026] 3. Flow rate: 1.0mL / min, injection volume: 20μL, column temperature: 38℃;

[0027] 4. Detection wavelength: 260nm, scanning wavelength: 200-600nm;

[0028] 5. Preparation of 20-hydroxyecdysterone (20E; 20-Hydroxyecdysone), α-ecdysone (E; Ecdysone), 2-deoxy-20-hydroxyecdysone (2D-20H, 2-deoxy-20- Hydroxyecdysone) and 7-dehydrocholesterol (7-DHC, 7-Dehydrocholesterol) standard solutions and mixed stand...

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Abstract

The invention discloses a method applying HPLC and simultaneously determining phytoecdysone substance; the method comprises the steps of: a, using YMC 250X4.6mm, S-5um chromatographic column; b, using methanol-water mobile phase to carry out gradient wash out; C, flow velocity: 1.0mL/min, sample size: 20uL, column temperature: 38 DEG C; d, wavelength: 260nm, scanning wavelength: 200-600nm. The method is simple in operation, fast in splitting, high in flexibility, the chromatographic column needs not to be changed, and the result is good in stability and high in repeatability.

Description

technical field [0001] The present invention belongs to the field of analytical chemistry. The invention relates to a chromatographic detection method, in particular to a detection method for simultaneous determination of plant ecdysone substances by using high performance liquid chromatography. Background technique [0002] Ecdysone is a very important class of plant secondary metabolites, a natural steroidal compound with insect molting activity, and plays a very important role in plant resistance to herbivorous insects. Therefore, elucidating the biosynthetic pathway of plant ecdysone is not only a theoretical breakthrough in basic research, but also provides a new technology for agricultural pest control in production. [0003] In the pathway of plant ecdysone biosynthesis, 20-hydroxyecdysterone is its final product, 7-dehydrocholesterol, 2-deoxy-20-hydroxyecdysterone and α-ecdysterone are intermediate products, all of the above compounds It has a 7-en-6-one structure ...

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Application Information

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IPC IPC(8): G01N30/02
Inventor 宋雯雯段方猛
Owner QINGDAO AGRI UNIV
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