Pharmacokinetic Study on Piplartine and Piperine after Oral Administration of Piper chaba Root by Liquid Chromatography‑mass Spectrometry/mass Spectrometry

Background: Piperaceae family are a well‑known source of structurally diverse amides with the wide range of bioactivities such as cytotoxic, stomach aches, insect repellents, anti‑inflammatory, insecticidal, and antifeedant activities. It has been reported that piplartine and piperine, alkaloid/amide compounds from Piper species, show antitumor activities. Objective: A rapid, sensitive liquid chromatography‑tandem mass spectrometry method has been developed and validated for the determination of piplartine and piperine from Piper chaba extract. Materials and Methods: The two analytes, together with internal standard (IS, trichostachine), were separated on a Waters Acquity ethylene bridged hybrid C18 (2.1 mm × 100 mm, 1.9 µ) column using a mobile phase of acetonitrile with 0.1% formic acid and water with 0.1% formic acid (70:30, v/v) with isocratic elution. The detection was performed using the positive ion electrospray ionization in multiple reaction monitoring mode with transitions at m/z 318→221 for piplartine, m/z 286→201 for piperine, and m/z 272→201 for the IS. Results: The calibration curves were both linear (r2 > 0.995) over a concentration range of 1.0–2000 ng/mL; the lower limit of detection quantification was 1.0 ng/mL for both piplartine and piperine. The intra‑day and inter‑day precisions (relative standard deviation %) were <10.9%, and recoveries ranged from 90.3% to 103.0%. Conclusions: The analytes were proven stable in the short‑term, long‑term, and after three freeze‑thaw cycles. The method was successfully applied to pharmacokinetic studies of piplartine and piperine in rats after oral administration of P. chaba extract.