Development and Validation of Analytical Method for Simultaneous Estimation of Bupivacaine and Meloxicam in Human Plasma Using UPLC-MS/MS
Bupivacaine HCl (BP) and Meloxicam (ME) are used as intra-operative local anesthesia and non-steroidal anti-inflammatory drugs respectively. A simple, rapid, selective and sensitive ultra pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for simultaneous quantification of BP and ME in human plasma. By following a simple protein precipitation with solvent mixture containing Water:Acetonitrile:Formic Acid (76:24:0.1,v/v/v), the analytes and internal standards were separated on Acquity HSS T3 column (2.1 x 50 mm, 1.8 μm). Chromatographic Mobile phase A consist of 10 mM Ammonium Formate and Mobile phase B consist of a mixture of Acetonitrile:water:Formic acid (96:5:0.2, v/v/v). Bupivacaine-d9 (BUd9) used as internal standard for BP and 13C,d3 Meloxicam (ME13cd3) used for ME. The concentration of BP and ME were determined by gradient UPLC-MS/MS in Multiple reaction mode (MRM). The mass transition ion-pair for BP was m/z 289.3→140.0 and BUd9 was 298.3→ 149.0, for ME was m/z 352.1→115.0 and ME13cd3 was 356.1→115.0, respectively. The proposed method was linear in the range 10 - 4500 ng/mL. The intra and inter-run precision values are within 6.8 % at a lower limit of quantization level. The overall recovery for both analytes found 98%. The total run time of the method is 2.5 min.