title: Nateglinide quantification in rabbit plasma by HPLC: optimization and application to pharmacokinetic study.
abstract: A rapid, simple, and sensitive HPLC method with UV detection was developed and validated for the determination of nateglinide (NTG) from rabbit plasma. The retention behavior of NTG and gliclazide (GLZ, internal standard-IS) as a function of mobile phase pH, composition and flow rate was investigated. Separation was developed on a reverse-phase C(18) column (250 mm x 4.6mm i.d., 5 microm particle size), using a mixture of acetonitrile (ACN):10mM phosphate buffer (PBS, pH 3.0) in the ratio of 70:30(%v/v) at a flow rate of 1.0 ml/min with UV detection at 203 nm within 8 min, and quantified based on drug/IS peak area ratios. The plasma samples were prepared by a simple deproteinization with a mixture of methanol and acetonitrile, yielding more than 97.86% extraction efficiencies. The calibration curve was linear (correlation coefficient of 0.9984) in the concentration range of 10-2500 ng/ml. The limit of detection (LoD) and limit of quantitation (LoQ) were found to be 2.91 and 9.70 ng/ml, respectively. Both the intra-day and inter-day precisions at four tested concentrations were below 1.32% R.S.D. The present method was selective enough to analyze NTG in rabbit plasma without any tedious sample clean-up procedure and was successfully applied for estimating the pharmacokinetic parameters of NTG following oral administration of a single 15 mg NTG to white albino rabbits.
authors.affiliation: Center of Relevance and Excellence in Novel Drug Delivery Systems, Pharmacy Department, The M.S. University of Baroda, Vadodara-390 002, India. jollymayur@hotmail.com
authors.forename: Jolly M
authors.initials: JM
authors.lastname: Sankalia
authors.forename: Mayur G
authors.initials: MG
authors.lastname: Sankalia
authors.forename: Vijay B
authors.initials: VB
authors.lastname: Sutariya
authors.forename: Rajashree C
authors.initials: RC
authors.lastname: Mashru
mesh_terms: D000284:Administration, Oral; D000818:Animals; D019540:Area Under Curve; D002021:Buffers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D003080:Cold Temperature; D003510:Cyclohexanes / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D004355:Drug Stability; D005615:Freezing; D006207:Half-Life; D006863:Hydrogen-Ion Concentration; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D008657:Metabolic Clearance Rate; D015394:Molecular Structure; D000077715:Nateglinide; D010649:Phenylalanine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D011817:Rabbits; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D013997:Time Factors
chemical_list: D002021:Buffers; D003510:Cyclohexanes; D007004:Hypoglycemic Agents; D000077715:Nateglinide; D010649:Phenylalanine