Abstract

RAPID DETERMINATION OF CIPROFLOXACIN CONCENTRATION IN HUMAN PLASMA BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Reem Alswayeh, Rajaa F. Hussein, Syed N. Alvi and Muhammad M. Hammami*

ABSTRACT

A simple, precise, and rapid high performance liquid chromatography (HPLC) method for the determination of ciprofloxacin level in human plasma using gatifloxacin as an internal standard (IS) was developed and validated. 0.5 ml Plasma samples containing ciprofloxacin were mixed with 200 μl of IS (50 μg/ ml in buffer) and filtered through a centrifugal filter device by centrifuging at 4000 rpm for 30 minutes. A 100 μl of ultra-filtrate clear solution was injected in the HPLC system. The compounds of interest were efficiently separated on Xterra RP-18 (4.6 x 150 mm, 5-μm) steel column (25ºC) proceeded by a universal sentry guard column (symmetry C18, 5-μm insert) and detected with a photodiode array detector set at 276 nm. The mobile phase consisted of a mixture of 0.025 M of sodium phosphate monobasic (pH = 3.0, adjusted with phosphoric acid) and acetonitrile (85:15, v:v). The mobile phase was delivered at a flow rate of 1 ml/min and run for 10 min. No interference in blank plasma or by commonly used drugs was observed, and the detection limit of ciprofloxacin was 0.05 μg/ml. The relationship between ciprofloxacin concentration in plasma and peak area ratio of ciprofloxacin /IS was linear (R2 ≥ 0.9997) in the range of 0.1 – 12.0 μg/ml. Intra- and inter-day coefficient of variation and bias were ≤ 7.0% and ≤ 11.1%, respectively. Mean extraction recovery of ciprofloxacin and the IS from the plasma samples were ≥ 90%, and 85%, respectively. Using the method, ciprofloxacin in found to be stable in human plasma for 16 weeks at -20 ºC. Further, the method was successfully employed to measure ciprofloxacin level in plasma samples from a healthy volunteer.

Keywords: Ciprofloxacin, Gatifloxacin, Human plasma, HPLC.