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Abstract

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD WITH FLUORESCENCE DETECTION FOR SULPIRIDE DETERMINATION IN HUMAN PLASMA

*Mohammed Kaleemullah, Yusrida Darwis, Peh Kok Yang, Jiyauddin Khan ,Mallikarjun Chitneni, Fadli Asmani and Eddy Yusuf

ABSTRACT

The present study was aimed to develop and validate a simple reversed-phase high performance liquid chromatographic method for determination of sulpiride in human plasma. The HPLC system was operated at an excitation and emission wavelengths of 300 nm and 365 nm, respectively with the gain was set at 4 and sensitivity at medium. The mobile phase was consisted of 0.01 M phosphoric acid, acetonitrile and methanol (84:12:4, v/v) with addition of Triethylamine (0.15%v/v). The mobile phase pH was adjusted to 6 by using glacial acetic acid. The mobile phase was isocratically pumped at a flow rate of 1 mL/min. The analytical column Luna C18 (5 m, 250 x 4.6 mm ID, Phenomenex, USA) fitted with a refillable guard column (Upchurch Scientific, Oak Harbour, WA, USA) packed with Perisorb RP-18 (30-40 m, pellicular) was used for chromatographic separation. The mobile phase was filtered under vacuum through 0.45 μm nylon membrane filter (Whatman International, England) and degassed before used. The calibration curve was linear in the range of 15 to 4000 ng/ml with correlation coefficient (r) of 0.9999 (± 0.0001). The intra-day accuracy ranged from -4.59% to 12.91% with a precision from 1.42% to 6.79%. The inter-day accuracy ranged from -1.86% to 6.29% with a precision from 4.21% to 13.91%. The extraction recovery values were found to be 95.99 ± 9.44%, 96.12 ± 11.94% and 93.49 ± 5.13%, with precision of 9.84%, 12.42% and 5.49% respectively. The mean recovery for internal standard (metoclopramide) was 90.28 ± 3.95%. The values of accuracy, precision and recovery obtained were within the acceptable limits as proposed by USFDA Bioanalytical Guidelines. In conclusion, a simple, accurate, precise, sensitive, and specific RP-HPLC method was developed for the determination of sulpiride in human plasma.

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