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.
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[0022] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.
[0023] A detection method for simultaneous determination of phytoecdysone substances by high performance liquid chromatography, comprising the following steps:
[0024] 1. Adopt YMC250×4.6mm, S-5μm chromatographic column;
[0025] 2. Use methanol-water mobile phase for gradient elution, the gradient elution program 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°C;
[0027] 4. Detection wavelength: 260nm, scanning wavelength: 200-600nm;
[0028] 5. Prepare 20-hydroxyecdysone (20E; 20-Hydroxyecdysone), α-ecdysone (E; Ecdysone), 2-deoxy-20-hydroxyecdysone (2D-20H, 2-deoxy-20- Hydroxyecdysone) and 7-dehydrocholesterol (7-DHC, 7-Dehydrocholesterol) s...
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