Development of Liquid ChromatographyÃ¢ÂÂUV Method for Simultaneous Determination of Leflunomide and NSAIDs in API and Pharmaceutical Formulations: It Ã¢ÂÂ s Application to In vitro Interaction StudiesNajma Sultana1, Mohammed Saeed Arayne2*, Moona Mehboob Khan1 and Saeeda Nadir Ali2
- *Corresponding Author:
- Mohammed Saeed Arayne
Department of Chemistry, University of Karachi
Karachi 75270, Pakisthan
E-mail: [email protected]
Received date: June 29, 2013; Accepted date: September 12, 2013; Published date: September 15, 2013
Citation: Sultana N, Arayne MS, Khan MM (2013) Development of Liquid Chromatography–UV Method for Simultaneous Determination of Leflunomide and NSAIDs in API and Pharmaceutical Formulations: It’s Application to In vitro Interaction Studies. Med chem 3:262-270. doi:10.4172/2161-0444.1000149
Copyright: © 2013 Sultana N, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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.