RAPID AND VALIDATED HPLC-UV METHOD FOR DETERMINATION OF GEMIFLOXACIN IN HUMAN URINE

Objective: To develop and validate a simple and rapid reversed phase high performance liquid chromatographic (RP-HPLC) method for the determination of Gemifloxacin (GFX) in human urine.Methods: GFX was isolated from urine samples after acidification using methylene chloride. Good chromatographic separation was achieved using C18 Ultrasphere (250 mm × 4.6 mm, 5 µm.) analytical column maintained at 25 °C. The mobile phase consisted of methanol and 0.1 M phosphate buffer pH 3 in the ratio of (48: 52, v/v), respectively. The analysis time was 10 min at a 1.0 ml/min flow rate. The UV detection was carried out at 272 nm.Results: GFX has been eluted at 7.5 min. Linearity was obtained over a concentration range of 20-200 ng/ml (r2>0.999). The extraction recovery of GFX from urine samples was 60%. The proposed method demonstrated excellent intra-and inter-day precision and accuracy within 1.19% and 100.65 %, respectively. The limit of detection (LOD) was found to be 6.3 ng/ml.Conclusion: Simple and accurate RP-HPLC method for determination of GFX in human urine was developed and validated. The method was successfully applied for determination of GFX in human urine samples from healthy volunteers up to 24 hours after oral administration of 320 mg gemi floxacin tablets.Â

Determination of Atenolol and Trimetazidine in Pharmaceutical Tablets and Human Urine Using a High Performance Liquid Chromatography-Photo Diode Array Detection Method

A simple RP-HPLC-PDA method for determination of atenolol (ATN) and trimetazidine (TMZ) in human urine and tablets has been developed. Analytes were separated on a Caltrex BI column (125× 4.0 mm, 5 μm) with 25mM potassium dihydrogen phosphate pH 3.3, methanol, and acetonitrile mobile phases. The PDA detector was operated at 210 nm for TMZ and 225 nm for ATN and the flow rate was 1.0 mL/ min. Linearity was obtained over a concentration range of (1.0-100 μg/mL) for both analytes in standard solutions and the method was successfully applied for determination of target analytes in their pharmaceutical tablets. Excellent linearity was also obtained over concentration ranges of (0.25-25 μg/mL) and (0.5-25 μg/mL) in human urine for TMZ and ATN, respectively. A simple liquid-liquid extraction was applied for urine sample clean-up and a gradient method was used for chromatographic separation. The lower limit of quantitation (LOQ) was 0.99 and 0.60 μg/mL for ATN and TMZ, respectively. The limit of detection (LOD) was 0.30 and 0.18 μg/mL for ATN and TMZ, respectively. Inter- and intraday precision and accuracy for ATN were within ±1.89% in pure form and within ±2.85% in urine samples. Inter- and intraday precision and accuracy for TMZ were within ± 3.99% in pure form and within ± 3.19% in urine samples