Objective: The aim of the work was to develop and validate a simple, sensitive and selective Liquid chromatography with Mass spectroscopic method for simultaneous quantification of lidocaine and prilocaine in human plasma.
Design and methods: Analytes and the internal standards from human plasma were extracted by using solid- phase extraction technique using Waters Oasis® HLB 1 cc (30 mg) cartridges. The reconstituted samples were chromatographed on Phenomenex Kinetex EVO 4.6*100 mm 2.6 μ 100A column by using a mixture of acetonitrile and 5 mM ammonium acetate buffer (80:20, v/v) as the mobile phase at a flow rate of 0.6 mL/min.
Results: The method was validated over the concentration range of 0.10-201.80 ng/mL for lidocaine and 0.10-201.66 ng/mL for prilocaine. The calibration curve obtained was linear.
Conclusion: Method validation was performed as per FDA guidelines and the results met the acceptance criteria. A run time of 3.0 min for each sample, make it possible to analyze more than 350 human plasma samples per day. The proposed method was found applicable for pharmacokinetic studies.
Keywords: Human plasma; LC-MS/MS; Lidocaine; Pharmacokinetics; Prilocaine; Solid-phase extraction.