title: Development and validation of an LC-MS/MS method for simultaneous determination of piperaquine and 97-63, the active metabolite of CDRI 97-78, in rat plasma and its application in interaction study.
abstract: Piperaquine-dihydroartemisinin combination is the latest addition to the repertoire of ACTs recommended by the World Health Organization (WHO) for treatment of falciparum malaria. Due to the increasing resistance to artemisinin derivatives, CSIR-CDRI has developed a prospective short acting, trioxane antimalarial derivative, CDRI 97-78. In the present study, a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the simultaneous quantification of piperaquine (PPQ) and 97-63, the active metabolite of CDRI 97-78 found in vivo, was developed and validated in 100 μL rat plasma using halofantrine as internal standard. PPQ and 97-63 were separated using acetonitrile:methanol (50:50, v/v) and ammonium formate buffer (10 mM, pH 4.5) in the ratio of 95:5(v/v) as mobile phase under isocratic conditions at a flow rate of 0.65 mL/min on Waters Atlantis C18 (4.6 × 50 mm, 5.0 µm) column. The extraction recoveries of PPQ and 97-63 ranged from 90.58 to 105.48%, while for the internal standard, it was 94.27%. The method was accurate and precise in the linearity range 3.9-250 ng/mL for both the analytes, with a correlation coefficient (r) of ≥ 0.998. The intra- and inter-day assay precision ranged from 2.91 to 8.45% and; intra- and inter-day assay accuracy was between 92.50 and 110.20% for both the analytes. The method was successfully applied to study the effect of oral co-administration of PPQ on the pharmacokinetics of CDRI 97-78 in Sprague-dawley rats and vice versa. The co-administration of CDRI 97-78 caused significant decrease in AUC0-∞ of PPQ from 31.52 ± 2.68 to 14.84 ± 4.33 h*µg/mL. However, co-administration of PPQ did not have any significant effect on the pharmacokinetics of CDRI 97-78.
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR- Central Drug Research Institute, Lucknow, Uttar Pradesh, India.|Academy of Scientific and Industrial Research, New Delhi, India.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Wahajuddin
authors.affiliation: Analytical Chemistry Division, CSIR-Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India.
authors.forename: Sheelendra Pratap
authors.initials: SP
authors.lastname: Singh
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR- Central Drug Research Institute, Lucknow, Uttar Pradesh, India.|Academy of Scientific and Industrial Research, New Delhi, India.
authors.forename: Isha
authors.initials: I
authors.lastname: Taneja
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR- Central Drug Research Institute, Lucknow, Uttar Pradesh, India.|Academy of Scientific and Industrial Research, New Delhi, India.
authors.forename: Kanumuri Siva Rama
authors.initials: KS
authors.lastname: Raju
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR- Central Drug Research Institute, Lucknow, Uttar Pradesh, India.|Academy of Scientific and Industrial Research, New Delhi, India.
authors.forename: Jiaur Rahman
authors.initials: JR
authors.lastname: Gayen
authors.affiliation: Faculty of Pharmacy, Integral University, Lucknow, India.
authors.forename: Hefazat Hussain
authors.initials: HH
authors.lastname: Siddiqui
authors.affiliation: Pharmacokinetics and Metabolism Division, CSIR- Central Drug Research Institute, Lucknow, Uttar Pradesh, India.|Academy of Scientific and Industrial Research, New Delhi, India.
authors.forename: Shio Kumar
authors.initials: SK
authors.lastname: Singh
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000097:blood* / Q000493:pharmacokinetics; D019540:Area Under Curve; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000097:blood* / Q000493:pharmacokinetics*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004347:Drug Interactions; D007202:Indicators and Reagents; D011446:Prospective Studies; D011786:Quality Control; D011804:Quinolines / Q000097:blood* / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D013141:Spiro Compounds / Q000097:blood* / Q000493:pharmacokinetics; D013343:Styrenes / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry; D010879:Piperazines
chemical_list: D000962:Antimalarials; D019086:Bridged Bicyclo Compounds, Heterocyclic; D007202:Indicators and Reagents; D011804:Quinolines; D013141:Spiro Compounds; D013343:Styrenes; C000611963:CDRI 97-63; C574493:CDRI-97-78; C034759:piperaquine; D010879:Piperazines
keywords: LC-MS/MS; drug interaction; malaria; recovery; validation