Rosiglitazone quantification in rat plasma using high performance liquid chromatography with mass spectrometric detection and its application to pre-clinical pharmacokinetic studies.

Unlabelled: A rapid and sensitive high performance liquid chromatography-mass spectrometry method has been developed for quantitative determination of Rosiglitazone, a thiazolidinedione drug used for the treatment of type II diabetes mellitus in rat plasma. The method was also validated as per International Conference on Harmonization (ICH) and Food and Drug Administration (FDA) guidelines. The analyte was extracted from rat plasma by the simple precipitation of plasma proteins technique using acetonitrile as a precipitating agent. Alprazolam was used as the internal standard. A Chromolith RP-18(e) column provided chromatographic separation of the analyte using a mobile phase containing 5mM ammonium acetate in water (pH 3) and methanol (20:80) at a flow rate of 1 mL/min with an elution time as low as 2.5 min, which was followed by detection with mass spectrometry. The mass transition ion-pair was followed as m/z 358.0 for rosiglitazone and m/z 308.8 for alprazolam. Simple isocratic chromatographic conditions and mass spectrometric detection of the method enable the detection of rosiglitazone at less than nanogram levels. The proposed method was found to be linear from 0.5 to 100 ng/mL (r(2) = 0.9987). The percent coefficient of variance for precision and accuracy values found at LLOQ (9.17), LQC (8.45), MQC (1.66) and HQC (1.53). The overall recovery of rosiglitazone was 95.9%. The developed and validated method was successfully applied for the pharmacokinetic studies of rosiglitazone tablets after a single oral dose to healthy Wistar rats.

Lay Abstract: A rapid and sensitive high performance liquid chromatography-mass spectrometry method has been developed and validated for quantification of rosiglitazone in rat plasma. The analyte was extracted from rat plasma by simple precipitation technique. Alprazolam was used as the internal standard. A Chromolith RP-18(e) column provided chromatographic separation of the analyte using a mobile phase containing 5mM ammonium acetate in water and methanol (20:80) at a flow rate of 1 mL/min which was followed by detection with mass spectrometry. The mass transition ion-pair was followed as m/z 358.0 for rosiglitazone and m/z 308.8 for alprazolam. The method involves precipitation of rosiglitazone from plasma, simple isocratic chromatography conditions and mass spectrometric detection that enables detection at less than nanogram levels. The proposed method has been validated with a linear range of 0.5 to100 ng/mL for rosiglitazone. The percent coefficient of variation for precision and accuracy are within 10%. The overall recovery of rosiglitazone was 95.9%. Total elution time was as low as 2.5 min. The developed and validated method was successfully applied for the pharmacokinetic studies of rosiglitazone tablets after a single oral dose to rats.