Bioanalytical Method Validation for Determination of Aceclofenac in K2EDTA Human Plasma by LC-MS/MS

R. Subramanian1, M. Purushothaman*2
1Sun Rise University, Alwar, Rajasthan – 301030, India
2Scient Institute of Pharmacy, Ibrahimpatnam, Ranga Reddy (Dt), Hyderabad, Talangana-501 506, India.

A B S T R A C T
A simple reverse phase liquid chromatographic and mass spectroscopic analytical method has been developed and validated for estimation of Aceclofenac in plasma. The separation was carried out on Hypersil Gold C18,100 X 2.1 mm, 1.9 µ as Stationary phase, Mobile Phase: Methanol: 10mm ammonium acetate buffer pH 6.8 Elution mode : Isocratic A: B= 80:20% v/v Flow rate: 350 µL/min.  Diclofenac was used as internal standard. The Aceclofenac and Diclofenac showed retention factor of 0.8 min ± 0.2 min respectively. The injection volume was 5µL and the total run time was 2 min. The method shows selectivity and linearity. The described LC-MS/MS method was linear over a concentration range of 38.673 to 19800.30 ng/mL. The extraction recoveries for Aceclofenac and Diclofenac were found to be between 69.11 and 72.15%. The method shows to be stable for the studied parameters. The stability of the drug spiked human plasma samples during three freeze thaw cycles were stable in plasma for about one month when stored at frozen state. The results of the study showed that the proposed LC-MS/MS method is simple, rapid, precise and accurate, which is useful for the estimation of Aceclofenac in bulk fluids and biological plasma sample analyte with accuracy and reproducibility.
Keywords: Aceclofenac, LC MS method, Diclofenac and Freeze thaw cycles