Authors: Jun Zhe Min Suguru Hatanaka Toshimasa Toyo’oka Shinsuke Inagaki Ruri KikuraHanajiri Yukihiro Goda
Publish Date: 2009/09/08
Volume: 395, Issue: 5, Pages: 1411-
Abstract
A simultaneous determination method based on ultraperformance liquid chromatography UPLC with fluorescence FL detection and electrosprayionization timeofflight mass spectrometry ESITOFMS was developed for 16 “designated substances” ShiteiYakubutsu controlled by the Pharmaceutical Affairs Law in Japan These substances were first labeled with 4NNdimethylaminosulfonyl7fluoro213benzoxadiazole at 60 °C for 2 h in 01 M borax pH 93 The resulting fluorophores were well separated by reversedphase chromatography using an Acquity UPLC™ BEH C18 column 17 μm 100 mm × 21 mm id by isocratic elution with a mixture of water and acetonitrile–methanol 2080 containing 01 formic acid The separated derivatives were sensitively detected by both FL and TOFMS However the determination of several designated substances by FL detection showed interference from endogenous substances in biological samples Therefore the determination in real samples was carried out by a combination of UPLC separation and ESITOFMS detection The structures of the designated substances were identified from the protonatedmolecular ions M+H+ obtained from the TOFMS measurement The calibration curves obtained from the peak area ratios of the internal standard IS ie 3phenyl1propylamine and the designated substances versus the injection amounts showed good linearity The limits of detection left textS/N = 3 right and the limits of quantification left textS/N = 10 right in 01 mL of human plasma and urine for the present method were 030–150 pmol and 10–500 pmol respectively Good accuracy and precision according to intraday and interday assays were also obtained with the present procedure This method was applied to analyses of human plasma urine and real products
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