title: Liquid chromatographic determination of CPZEN-45, a novel anti-tubercular drug, in biological samples.
abstract: CPZEN-45 is a new drug candidate being considered for the treatment of tuberculosis (TB). The aim of this study was to develop and validate a reverse-phase high-performance liquid chromatographic (HPLC) method suitable to determine CPZEN-45 concentrations in biological samples. CPZEN-45 was extracted from biological fluids and tissues (plasma, lung and spleen from guinea pig) by sequential extraction with acetonitrile and quantified by a Waters HPLC Alliance System coupled with a ZORBAX Bonus-RP column, guard column and UV detection at 263nm. The mobile phase was 20:80 acetonitrile:ultrapure-water with 0.05% TFA. The CPZEN-45 peak was eluted at 5.1min with no interference from the inherent peaks of plasma, bronchoalveolar lavage (BAL), lung or spleen tissues. Recovery of CPZEN-45 from biological samples was >96% of the spiked amount. The limit of detection was 0.05μg/ml and the limit of quantitation was 0.29μg/ml which was more than 5 and 21 times lower than the reported minimal inhibitory concentration of CPZEN-45 (MIC=1.56μg/ml for Mycobacterium tuberculosis and 6.25μg/ml for MDR-TB, respectively). Thus, HPLC method was deemed reliable, sensitive, reproducible and accurate for the determination of CPZEN-45 concentrations in plasma, BAL, lung and spleen tissues. Therefore, this method was used in subsequent studies in the guinea pig model to determine the disposition of CPZEN-45 after administration of solutions by the IV and SC routes.
authors.affiliation: Department of Pharmaceutical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73126-0901, United States. Electronic address: shumaila-hanif@ouhsc.edu.
authors.forename: S N M
authors.initials: SN
authors.lastname: Hanif
authors.forename: A J
authors.initials: AJ
authors.lastname: Hickey
authors.forename: L
authors.initials: L
authors.lastname: Garcia-Contreras
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000032:analysis* / Q000493:pharmacokinetics; D001381:Azepines / Q000032:analysis* / Q000493:pharmacokinetics; D001992:Bronchoalveolar Lavage Fluid; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D006168:Guinea Pigs; D007262:Infusions, Intravenous; D007279:Injections, Subcutaneous; D008168:Lung / Q000187:drug effects; D008297:Male; D009169:Mycobacterium tuberculosis / Q000187:drug effects; D015203:Reproducibility of Results; D012984:Software; D013154:Spleen / Q000187:drug effects; D014018:Tissue Distribution
chemical_list: D000995:Antitubercular Agents; D001381:Azepines; C583520:CPZEN-45
keywords: CPZEN-45; Guinea pig; HPLC; Tuberculosis; Validation