Analysis of 20-Hydroxyecdysone and Its Analogs by High Performance Liquid Chromatography – Mass Spectrometry

• Main Authors: Wei-Chiang Shiu, 徐偉強
• Other Authors: Huey-Fen Tzeng
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/36410057781492502114

碩士 === 國立暨南國際大學 === 應用化學系 === 105 === 20-Hydroxyecdysone and its analogs play important roles on the development of insects, particularly the regulation of molting, reproduction, stress response and life. The total amount of 20-hydroxyecdysone and its analogs can be estimated by enzyme-linked immunoassay; however, the concentration of each compound is not clear. Herein, one method for analysis of 20-hydroxyecdysone, ecdysone, cyasterone and ponasterone A using high performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometer (HPLC-APCI-MS) is developed. Four analytes and one internal standard were successfully separated in 23 minutes using reverse phase liquid chromatography, and their concentrations were determined by using APCI-MS in positive ion mode. The linear range of the method using makisterone A as an internal standard was 2 – 50 μM. This method can be applied to analyze the composition of 20-hydroxyecdysone and its analogs in Drosophila melanogaster (2 days). Only ponasterone A (1040±70 pg/mg ) and 20-hydroxyecdysone (74.3±0.4 pg/mg) were detected; however, cyasterone and α-ecdysone were below limit of detection. It demonstrated that this method is successfully applied to analyze the composition of 20-hydroxyecdysone and its analogs in biological samples.