A simultaneous determination method for 5-fluorouracil and its metabolites in human plasma with linear range adjusted by in-source collision-induced dissociation using hydrophilic interaction liquid chromatography-electrospray ionization-tandem mass spectrometry

Biomed Chromatogr. 2016 Nov;30(11):1882-1886. doi: 10.1002/bmc.3743. Epub 2016 May 15.

Abstract

We applied a new technique for quantitative linear range shift using in-source collision-induced dissociation (CID) to complex biological fluids to demonstrate its utility. The technique was used in a simultaneous quantitative determination method of 5-fluorouracil (5-FU), an anticancer drug for various solid tumors, and its metabolites in human plasma by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS). To control adverse effects after administration of 5-FU, it is important to monitor the plasma concentration of 5-FU and its metabolites; however, no simultaneous determination method has yet been reported because of vastly different physical and chemical properties of compounds. We developed a new analytical method for simultaneously determining 5-FU and its metabolites in human plasma by LC/ESI-MS/MS coupled with the technique for quantitative linear range shift using in-source CID. Hydrophilic interaction liquid chromatography using a stationary phase with zwitterionic functional groups, phosphorylcholine, was suitable for separation of 5-FU from its nucleoside and interfering endogenous materials. The addition of glycerin into acetonitrile-rich eluent after LC separation improved the ESI-MS response of high polar analytes. Based on the validation results, linear range shifts by in-source CID is the reliable technique even with complex biological samples such as plasma. Copyright © 2016 John Wiley & Sons Ltd.

Keywords: 5-fluorouracil; LC/ESI-MS/MS; human plasma; in-source CID; linear range shift.

Publication types

  • Validation Study

MeSH terms

  • Antimetabolites, Antineoplastic / blood*
  • Antimetabolites, Antineoplastic / metabolism*
  • Chromatography, High Pressure Liquid / methods*
  • Fluorouracil / blood*
  • Fluorouracil / metabolism*
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Limit of Detection
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Tandem Mass Spectrometry / methods

Substances

  • Antimetabolites, Antineoplastic
  • Fluorouracil