Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Liquid Chromatography Quadrupole Time-Of-Flight Tandem Mass Spectrometry for Selective Determination of Usnic Acid and Application in Pharmacokinetic Study

  • Fang, Minfeng (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University) ;
  • Wang, Hui (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University) ;
  • Wu, Yang (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University) ;
  • Wang, Qilin (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University) ;
  • Zhao, Xinfeng (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University) ;
  • Zheng, Xiaohui (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University) ;
  • Wang, Shixiang (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University) ;
  • Zhao, Guifang (Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University)
  • Received : 2013.01.28
  • Accepted : 2013.03.13
  • Published : 2013.06.20
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Abstract

A rapid and sensitive method for determining usnic acid of Lethariella cladonioides in rat was established using high performance liquid chromatography (HPLC) quadrupole time-of-flight (QTOF) tandem mass (MS/MS). Rat plasma was pretreated by mixture of acetonitrile and chloroform to precipitate plasma proteins. Chromatographic separation was achieved on a column ($50{\times}2.1$ mm, $5{\mu}m$) with a mobile phase consisting of water (containing $5{\times}10^{-3}$ M ammonium formate, pH was adjusted to 3.0 with formic acid) and acetonitrile (20:80, v/v) at a flow rate of 0.3 mL/min. A tandem mass spectrometric detection with an electrospray ionization (ESI) interface was conducted via collision induced dissociation (CID) under negative ionization mode. The MS/MS transitions monitored were m/z 343.0448 ${\rightarrow}$ m/z 313.2017 for usnic acid and m/z 153.1024 ${\rightarrow}$ m/z 136.2136 for protocatechuic acid (internal standard). The linear range was calculated to be 2.0-160.0 ng/mL with a detection limit of 3.0 pg/mL. The inter- and intra-day accuracy and precision were within ${\pm}7.0%$. Pharmacokinetic study showed that the apartment of usnic acid in vivo confirmed to be a two compartment open model. The method was fully valid and will probably be an alternative for pharmacokinetic study of usnic acid.

Keywords

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