Development of a selective and sensitive LC–MS/MS method for the quantification of α-asarone in mouse plasma and its application to pharmacokinetic studies

• Published in: Journal of pharmaceutical and biomedical analysis 2018-03, Vol.151, p.284-290
• Main Authors: Ramalingam, Prakash, Ganesan, Palanivel, Choi, Dong-Kug, Ko, Young Tag
• Format: Article
• Language: English
• Subjects: Administration, Oral; Animals; Anisoles - administration & dosage; Anisoles - blood; Anisoles - pharmacokinetics; Biological Availability; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; LC–MS/MS; Limit of Detection; Male; Mice; Mice, Inbred BALB C; Mouse plasma; Pharmacokinetics; Reproducibility of Results; SLNs; Spectrometry, Mass, Electrospray Ionization - instrumentation; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry - instrumentation; Tandem Mass Spectrometry - methods; α-Asarone
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2018.01.024

•A method for quantifying α-asarone in mouse plasma is reported.•Simple protein precipitation method was utilized.•Small-volume serial blood sampling in mice method was employed.•Successful application in α-asarone loaded SLNs pharmacokinetic studies. A simple, sensitive and selective liquid chromatography-tandem mass spectrometric method was developed and validated for the quantification of α-asarone in mouse plasma with its application to pharmacokinetic studies. An electrospray ionization (ESI) with multiple reaction monitoring (MRM) mode was used to monitor the precursor-product ion transitions of 209.1 > 193.9 m/z for α-asarone and 157.8 > 114.0 m/z for allantoin. Chromatographic separation was acquired on a Sepax BR-C18 (5 μm, 120 Å 1.0 × 100 mm) column with an isocratic mobile phase consisting of methanol and 0.1% formic acid (80:20, v/v). The developed bioanalytical method was successfully validated according to the United States Food and Drug Administration (US FDA) guidelines for linearity, selectivity, accuracy, precision, recovery, matrix effect, and stability. The validated method was successfully applied to a pharmacokinetics study of α-asarone along with a combination of pharmacokinetic techniques, including small-volume serial blood sampling in mice, reducing drug doses and the number of animals used, using a simple protein precipitation method and less solvent consumption will enable its use in further bioequivalence studies.