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Determination of visnagin in rat plasma by liquid chromatography with tandem mass spectrometry and its application to in vivo pharmacokinetic studies.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2009-02-04)
Pattaraporn Vanachayangkul, Veronika Butterweck, Reginald F Frye
ABSTRACT

A rapid and sensitive method for the determination of visnagin in rat plasma was developed using liquid chromatography tandem mass spectrometry (LC-MS/MS). The plasma samples were processed by protein precipitation with methanol:acetonitrile (25:75) and warfarin was used as the internal standard. After vortex mixing and centrifugation, the supernatants were diluted with water (1:5) and then directly injected onto a Phenomenex Synergi Max RP column (75 mm x 2.0 mm ID, 4 microm) with isocratic elution at a flow rate of 0.2 mL/min. The mobile phase consisted of water and methanol (15:85, v/v) containing 0.1% formic acid and 5mM ammonium acetate. The total run time (injection to injection) was less than 4 min. Detection of the analytes was achieved using positive ion electrospray in the selected reaction monitoring (SRM) mode. The linear standard curve ranged from 1.0 to 5000 ng/mL and the precision and accuracy (inter- and intra-run) were within 4.5% and 4.3%, respectively. The method, which is rapid, simple, and precise, was successfully used to support a visnagin pharmacokinetics study.

MATERIALS
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Sigma-Aldrich
Khellin, for microscopy