LC-HRMS determination of piperine on rat dried blood spots: A pharmacokinetic study

Abstract

A liquid chromatography-high resolution mass spectrometry (LC-HRMS) method was developed and validated for the determination of piperine (PPR) on dried blood spots (DBS). DBS samples were prepared by spiking the whole blood with analyte to produce 30 µL of blood spots on specimen collection cards. Chromatographic separation was achieved on an Atlantis dC₁₈ column using acetonitrile and water (0.1% formic acid) (85:15, v/v) as mobile phase in an isocratic mode of elution at a flow rate of 0.75 mL/min. MS detection was carried out in electrospray positive ion mode for the target ions and monitored at m/z 286.1465 for PPR and 272.1303 for the internal standard (IS). The developed method exhibited a linear dynamic range over 0.01-2000 ng/mL for PPR on DBS. The overall extraction recovery of PPR from DBS was 92.5%. Influence of hematocrit and spot volume on DBS was also evaluated and found to be well within the acceptable limits. The method was successfully applied to pharmacokinetic studies of PPR in rats.

Keywords: Dried blood spot; LC–HRMS; Pharmacokinetics; Piperine; Trichostachine.

Fig. 1

Extraction of PPR from dried blood spots.

Fig. 2

Chemical structures of (A) PPR and (B) IS.

Fig. 3

ESI–HRMS spectra of (A) PPR and (B) IS.

Fig. 4

Representative LC–MS chromatograms of (A) a blank DBS and DBS samples spiked with IS and PPR at (B) LLOQ, (C) higher limit of quantification (HLOQ) and (D) 4.0 h after administration of a 15 mg/kg oral dose of PPR.

Fig. 5

Mean plasma concentration–time profile of PPR after administration of an oral dose of 15 mg/kg of PPR to male Wistar rats (Data are expressed as mean±SD (n=6)).