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2 2015

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Development and validation of simple, rapid and sensitive LC-PDA ultraviolet method for quantification of Nebivolol in rat plasma and its application to pharmacokinetic studies

Ravi P., Vats R., Joseph S., Gadekar N.

Acta Chromatographica

2015

Vol. 27, no. 2

281--294

A simple, sensitive and rapid high-performance liquid chromatography method with ultraviolet (UV) detector was developed and validated for the analysis of Nebivolol (NBL) in rat plasma. The plasma sample, spiked with raloxifene hydrochloride as an internal standard (IS), was subjected to single step protein precipitation method prior to analysis. Chromatographic separation was achieved on the Agilent C8 (150 mm × 4.6 mm, 5 μm) column and monitored at a wavelength of 280 nm. Elution was carried out, in an isocratic mode, using a mobile phase consisting of acetonitrile and potassium di-hydrogen orthophosphate buffer (pH 3.5 ± 0.1) in the ratio of 37:63 v/v. Retention times of IS and NBL were 5.1 ± 0.10 min and 8.01 ± 0.12 min, respectively. No interference was observed from plasma components in the analysis of NBL and IS. Calibration curve was linear over the range of 125–3000 ng/mL (r2 = 0.999). NBL was found to be stable under various processing and storage conditions. The developed method was applied in the quantification of NBL in plasma samples, determining various pharmacokinetic parameters from intravenous bolus and oral administration of the drug in Wistar rats. NBL was found to follow two compartmental open models in Wistar rats.

A Simple Liquid Chromatographic Method for the Determination of Tenofovir in Rat Plasma and Its Application to Pharmacokinetic Studies

Ravi P. R., Joseph S., Avula U. S. R., Anthireddy S.

Acta Chromatographica

2015

Vol. 27, no. 4

597--612

The objective of this study was to develop and validate a novel, simple, and selective high-performance liquid chromatographic (HPLC) method with photodiode array detector for the estimation of tenofovir in rat plasma, which can be utilized in analyzing the pharmacokinetic samples from rats. Prior to analysis, an optimized protein precipitation technique was used to extract tenofovir from plasma. The mobile phase for this method comprised of 10 mM ammonium acetate buffer (pH 4) and methanol in the ratio of 97:3 υ/υ. Chromatographic separation of tenofovir was achieved using Spincotech C-18G enabled column (250 × 4.6 mm, 5 μm). Tenofovir was monitored at a wavelength of 260 nm, and the calibration curve was linear in the range of 250–4000 ng mL−1 (R2 = 0.999). High recovery obtained after extraction (97%–101%) of plasma samples precluded the use of an internal standard. Validation studies were performed as per the standard guidelines, and the developed method was accurate, precise, and selective for the determination of tenofovir in the rat plasma. The stability studies performed during the sample pretreatment process and sample storage conditions did not show a quantifiable degradation of tenofovir. Further, this method was able to estimate tenofovir and determine its pharmacokinetic parameters, post IV bolus administration in male Wistar rats. The pharmacokinetic profile of tenofovir followed one compartmental open model.