Comparison of Electron Capture—Atmospheric Pressure Chemical Ionization and Electrospray Ionization for the Analysis of Gambogic Acid and its Main Circulating Metabolite in Dog Plasma

Abstract

Gambogic acid (GA), a promising anticancer candidate, is a polyprenylated xanthone abundant in the resin of Garcinia morella and Garcinia hanburyi. Electron capture-atmospheric pressure chemical ionization (EC- APCI) and electrospray ionization (ESI) techniques, both in the negative ion mode, were evaluated regarding ionization, fragmentation patterns and sensitivity for simultaneous liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of GA and its main circulating metabolite, 10-hydroxygambogic acid (10-OHGA) in dog plasma. Both analytes underwent extensive in-source fragmentation in EC-APCI, which was not desirable for reliable quantification of these analytes, whereas the substitution of ESI for EC-APCI almost eliminated the source instability of both analytes. Negative ion ESI was, therefore, chosen for the development of an LC-MS/MS method for simultaneous determination of these analytes. After protein precipitation by acetonitrile, all analytes were separated on a Luna C18 HST column (50 x 2.0 mm i.d., 2.5 microm) with a mobile phase of 20 mmol L(-1) ammonium acetate water solution containing 0.2% acetic acid:acetonitrile (18:82, v/v). The detection was performed on a tandem mass spectrometer using selective reaction monitoring mode. Calibration curves were linear over the range of 10-6000 ng mL(-1) for GA and 3-2000 ng mL(-1) for 10-OHGA. The method was successfully applied to the pharmacokinetics study of GA injection in six beagle dogs.