Simultaneous Determination of Podophyllotoxin, Quercetin and Kaempferol in Podophyllin by Liquid Chromatography Tandem Mass Spectrometry
MEI-CHIH LIN1,2, JER-HUEI LIN2, SHU-KONG CHEN2, YU-WEN CHENG1* AND HUEI-WEN CHENG1*
1. College of Pharmacy, Taipei Medical University, Taipei, Taiwan (R.O.C.)
2. Bureau of Food and Drug Analysis, Department of Health, Executive Yuan, Taipei, Taiwan (R.O.C.)
(Received: March 26, 2008; Accepted: September 5, 2008)
ABSTRACT
A quantitative high performance liquid chromatography-tandem mass spectrometry (LC/MS/MS) was developed to analyze the constituents of podophyllin. Chromatographic separation was performed on a Cosmosil 5C18-MS (25 cm × 4.6 mm I.D., 5 μm) reverse phase column using a gradient of mobile phase (0.25% formic acid-methanol). The column effluent was split 2: 3 into the photodiode detector and tandem mass spectrometer. Podophyllotoxin, quercetin and kaempferol in podophyllin were identified by daughter ion scan mode and then determined their contents by multiple-reaction monitoring (MRM) mode. The limits of detection and quantitation for podophyllotoxin, quercetin and kaempferol were 2.40, 8.01; 2.94, 9.87 and 3.10, 10.2 ng/mL, respectively. The relative standard deviations of intraday and interday analyses for podophyllotoxin, quercetin and kaempferol ranged from 0.52-6.01% and 2.48-9.88%, respectively. The mean recoveries for podophyllotoxin, quercetin and kaempferol were 101.4%, 98.3% and 98.7%, respectively. The developed LC/MS/MS method was suitable for the simultaneous determinations of podophyllotoxin, quercetin and kaempferol in podophyllin.
Key words: LC/MS/MS, podophyllin, podophyllotoxin, quercetin, kaempferol