Pharmacokinetic and metabolism studies of rohitukine in rats by high performance liquid-chromatography with tandem mass spectrometry.
A sensitive, selective, and rapid high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the quantification of rohitukine in rat plasma. HPLC was performed using a Symmetry-Shield C18 (5 μ, 4.6 × 150 mm) column, and isocratic elution with ammonium acetate buffer (pH4; 10 mM):methanol (08:92, v/v) at a flow rate of 0.6 mL/min. Sample clean-up involved solid phase extraction (SPE) of analyte and internal standard (phenacetin) from 100 μL plasma. The parent→product ion transitions (MRM) for analyte and IS were 306.1→245.1 m/z and 180.1→138.1 m/z respectively, and were monitored on a triple quadrupole mass spectrometer, operating in positive ion mode. The method was validated across the dynamic concentration range of 5-500 ng/mL for rohitukine, with a fast run time of 4.5 min. The analytical method measured concentrations of rohitukine with accuracy (% bias) of <±10% and precision (% RSD) of <±12%. Rohitukine was stable during the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and 30 days of storage in a freezer at -70±10°C. Finally, the applicability of this assay has been successfully demonstrated in vivo pharmacokinetic and in vitro metabolism studies in Sprague-Dawley rat. This method will therefore be highly useful for future preclinical and clinical pharmacokinetic studies of rohitukine.
Pharmacokinetics & Metabolism Div., CSIR - Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, Rafi Marg, New Delhi 110 001, India.
Yashpal S
YS
Chhonker
Pharmacokinetics & Metabolism Div., CSIR - Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, Rafi Marg, New Delhi 110 001, India.
Hardik
H
Chandasana
Pharmacokinetics & Metabolism Div., CSIR - Central Drug Research Institute, Lucknow 226031, India.
Deepak
D
Kumar
Medicinal & Process Chemistry Div., CSIR - Central Drug Research Institute, Lucknow 226031, India.
Sunil K
SK
Mishra
Medicinal & Process Chemistry Div., CSIR - Central Drug Research Institute, Lucknow 226031, India.
Shishir
S
Srivastava
Medicinal & Process Chemistry Div., CSIR - Central Drug Research Institute, Lucknow 226031, India.
Vishal M
VM
Balaramnavar
Pharmacology Div., CSIR - Central Drug Research Institute, Lucknow 226031, India.
Anil Nilkanth
AN
Gaikwad
Sophisticated Analytical Facility, CSIR - Central Drug Research Institute, Lucknow 226031, India.
Sanjeev
S
Kanojiya
Medicinal & Process Chemistry Div., CSIR - Central Drug Research Institute, Lucknow 226031, India.
Anil K
AK
Saxena
Pharmacokinetics & Metabolism Div., CSIR - Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, Rafi Marg, New Delhi 110 001, India. Electronic address: rabi.cdri@gmail.com.
Rabi S
RS
Bhatta
D061605:Administration, Intravenous; D000284:Administration, Oral; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002867:Chromones / Q000008:administration & dosage / Q000378:metabolism / Q000493:pharmacokinetics*; D008297:Male; D010880:Piperidines / Q000008:administration & dosage / Q000378:metabolism / Q000493:pharmacokinetics*; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
C573233:5,7-dihydroxy-2-methyl-8-(4-(3-hydroxy-1-methyl)-piperidinyl)-4H-1-benzopyran-4-one; D002867:Chromones; D010880:Piperidines
Bioanalytical method; Bioavailability; Dysoxylum binectariferum; LC–MS/MS; Pharmacokinetics; Rohitukine