An HPLC Method for the Determination of Granisetron in Guinea Pig Plasma*

Abstract

A high-performance liquid chromatographic (HPLC) method for the determination of granisetron (GRN) in guinea pig plasma has been developed. Guinea pig plasma spiked with GRN was microfiltered, and the recovered filtrate was directly injected onto the column without any further cleanup procedures. Separation was achieved on a spherical silica column and GRN was detected at 305 nm. Approximately 800-900 injections were made without any evidence of column deterioration. For the standard curves, correlation coefficients ranged from 0.9978-0.9999, and the percent standard deviation (%SD) from the mean area under the curve (AUC) was calculated to be less than 10% for all concentrations, except for the lowest concentration (0.325 ng/microL, 11.3%). Between-day and within-day coefficients of variation (%CV) ranged from 4.9 to 9.5% and 3.6 to 7.6%, respectively. Percent errors for within-day test plasma samples were not greater than 8.2% of the expected concentration for all samples except for 1.125 ng/microL (-14.6%). The limit of sensitivity was found to be 0.019 ng/microL. Estimated recovery of GRN in the microfiltrate was calculated to be 58-59% and 78-81% in plasma and water, respectively. Stability studies indicated that repeated refrigeration and warming (for six days) of microfiltered GRN plasma samples produced no changes in GRN concentrations from day to day. However, microfiltered GRN plasma samples that were repeatedly frozen and thawed demonstrated erratic concentration changes from day to day. The precision, accuracy, and small sample requirements of this method indicate its utility for pharmacokinetic studies in small animals where sample volume may be restrictive.

Rapid determination of granisetron in human plasma by liquid chromatography coupled to tandem mass spectrometry and its application to bioequivalence study

“…The results in Tables 1-3 show that this method is as accurate and precise as others reported in literature [12][13][14], while others have used higher volumes of plasma sample [10,15], or more laborious plasma extraction procedures [8], with similar results.…”

“…The analytical methods used to determine granisetron concentrations in biological samples include liquid chromatography (LC) with fluorescence [5][6][7][8][9][10][11][12], UV [13] and tandem mass spectrometric [14,15] detector. All the reported fluorescence or UV detection methods have inconveniences for routine analysis of large batches of biological samples for the reasons of large sample volumes, complex extraction procedures, inadequate sensitivity or long chromatographic run time.…”

Quantification of granisetron in human plasma by liquid chromatography coupled to electrospray tandem mass spectrometry

“…The bioanalytical component of a pharmacokinetic study requires a drug assay with simplicity, high sensitivity, selectivity, small volume requirements and rapid turnaround time. Only a few methods have been reported for the quantification of granisetron in biological fluids, which involves liquid chromatography (LC) with fluorescence detection (Boppana, 1995;Capacio et al, 1993;Cupissol et al, 1993;Huang et al, 1998Huang et al, , 1999Kudoh et al, 1994;Lazarus et al, 1999;Pinguet et al, Granisetron LC-ESI-MS/MS assay in human plasma 889 ORIGINAL RESEARCH 1996; Wada et al, 1998) and LC with tandem mass spectrometry (Boppana et al, 1996). In all the reported fluorescence detection methods, sensitivity was inadequate, chromatographic run time was longer, sample volume requirement was high and considered timeconsuming in sample preparation/extraction procedures.…”

First derivative spectrophotometric determination of granisetron hydrochloride in presence of its hydrolytic products and preservative and application to pharmaceutical preparations

“…The literature reveals that several methods have been reported for determination of GRS in pharmaceutical preparations and biological fluids. These include high performance liquid chromatography (HPLC) using DAD (diode‐array detection),4 fluorescence,5–11 ultra‐violet (UV),12, 13 tandem mass spectrometric,14–19 and electrochemical10 detectors. A flow‐injection chemiluminescence method has been reported for the determination of granisetron 20…”