Najma Sultana, Mohammed Saeed Arayne, Moona Mehboob Khan and Saeeda Nadir Ali
An efficient analytical method for the simultaneous determination of leflunomideand non steroidal anti-inflammatory drugs in API and formulations by LC-UV has been developed. The analytes were separated on Purospher Star, C18 (5 μm, 250×4.6 mm) column at ambient temperature with methanol: water (80:20, v/v, pH at 2.7) at flow rate of 1.5 mL min-1. Experiment was conducted in two phases. Leflunomide was separated with flurbiprofenand ibuprofen (phase- I) and diclofenac sodium and mefenamic acid (phase II). Calibration curves were linear over the range 0.625–5 μg mL-1 in both phases for leflunomide while for flurbiprofen, ibuprofen, diclofenac sodium, mefenamic acid linearity were achieved in the range of 0.625-5, 11.25-90, 1.56-50 and 0.78-25 μg mL-1, respectively with r2>0.9998. Intraday variation was <1.2 and <1.4 %, while in inter-day ranged between 0.042-1.45% and 0.08-1.27% in phase-I and II, respectively. Mean recovery values for intra-day ranged from 99.04-100.4% and 98.48-100.2% and for inter-day were between 98.54-100.29% and 98.85-100.54% in phase-I and II, respectively. The LLOD of leflunomide was 13 ng mL-1, while LLOQ was 39ng mL-1, respectively. LLOD and LLOQ for flurbiprofen, ibuprofen, diclofenac sodium and mefenamic acid were 6.9, 296, 71 and 1.2 ng mL-1 and 21, 897, 214.3 and 3.676 ng mL-1, respectively. Present study showed that nanogram quantities of all the compounds can be estimated accurately. The newly established method was successfully applied to study in vitro interactions between leflunomide and NSAIDs.
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