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Pharmacokinetic Study of Hypaphorine, a Potential Agent for Treating Osteoclast-based Bone Loss, on Rats Using
Taiyuan Zhang1, Yan Yan2, Yutao Xue3
1Department of Traumatic Orthopaedics, Affiliated Jinan Third Hospital of Jining Medical University, Jinan 250132, P.R. China.
Aims And Objectives:
A high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining hypaphorine, a potential agent for treating osteoclast- based bone loss, was developed and validated in rat plasma.
Materials And Methods:
Plasma samples were pretreated by the protein precipitation. Chromatographic separation was performed using an Inertsil ODS-3 column (50 mm × 4.6 mm, 5 μm). The mobile phase consisted of water (containing 0.1% formic acid) and acetonitrile in a gradient mode at a flow rate of 0.5 mL/min. The transitions from protonated precursor ion [M + H]+ to the particular daughter ion were acquired using selected reaction monitoring (SRM). The mass transitions of hypaphorine and IS were found to be 247 → 188 and m/z 219 → 188, respectively. The method was validated in terms of selectivity, linearity, accuracy and precision, extraction recovery and matrix effect, stability, and carryover.
Results:
It showed good linearity over the range of 1-2000 ng/mL (R2 = 0.9978). The intra-batch accuracy was within 93.95-105.81%, and the precision was within 4.92-11.53%. The inter-batch accuracy was within 96.18-100.39% with a precision of 6.22-11.23%. The extraction recovery and matrix factors were acceptable.
Conclusion:
The simple and rapid method was successfully applied to the pharmacokinetic study in rats following oral administration of hypaphorine at the doses of 0.5, 1.5, and 4.5 mg/kg.
