Validated HPLC method for determination of PAT-5A, an insulin sensitizing agent, in rat plasma

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 22; no. 2; pp. 251 - 255
• Main Authors: Mamidi, N.V.S.Rao, Kumar, V.V.S.Swaroop, Katneni, Kasiram, Chaluvadi, M.Rao, Shreeram, S, Gangadhar, S, Nataraj, B, Kishore, P.Hari, Bhushan, V, Subramaniam, S
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01-03-2000; Elsevier Science
• Subjects: Analysis; Animals; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; General and cellular metabolism. Vitamins; General pharmacology; Hypoglycemic Agents - blood; Hypoglycemic Agents - pharmacokinetics; Male; Medical sciences; PAT-5A-HPLC-method validation; Pharmacology. Drug treatments; Pyridines - blood; Pyridines - pharmacokinetics; Rats; Rats, Wistar; Reference Standards; Reproducibility of Results; Spectrophotometry, Ultraviolet; Thiazoles - blood; Thiazoles - pharmacokinetics; Thiazolidinediones; PAT-5A-HPLC-method validation; Biological fluid; Rat; Thiazole derivatives; Rodentia; Oral administration; HPLC chromatography; Bioavailability; Blood plasma; Vertebrata; Hypoglycemic agent; Mammalia; Absorption; Ultraviolet detector; Analysis method; Animal; Pharmacokinetics; Chemosensitizing agent; Quantitative analysis
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(99)00262-9

A high performance liquid chromatographic method for the determination of PAT-5A (a potent insulin sensitizer) using DRF-2095 (a thiazolidinedione) as internal standard (I.S.) is described. A 1:1 v/v ethylacetate and dichloromethane solvent mixture was used for extraction of PAT-5A from plasma. A Kromasil KR100-5C18-250A, 5 μm, 4.6×250 mm SS column was used for the analysis. Mobile phase consisting of sodium dihydrogen phosphate (pH 4.0, 0.05 M) and methanol mixture (25:75, v/v) was used at a flow rate of 1.0 ml/min. The eluate was monitored using a UV detector set at 345 nm. Ratio of peak area of analyte to I.S. was used for quantification of plasma samples. Using this method the absolute recovery of PAT-5A from rat plasma was >90% and the limit of quantification was 0.05 μg/ml. The intra-day relative standard deviation (RSD) ranged from 2.19 to 4.98% at 1.0 μg/ml, 1.05 to 3.68% at 10.0 μg/ml and 3.14 to 5.08% at 50 μg/ml. The inter-day RSD were 1.6, 2.24 and 1.54% at 1, 10 and 50 μg/ml, respectively. The method was applied to measure the plasma concentrations of PAT-5A in pharmacokinetic and bioavailability studies in male Wistar rats.