This research paper revolves around the process of the quantification of bupivacaine in human plasma through the selection of a sensitive high performance liquid chromatography method. The extraction of bupivacaine and ropivacaïne as an internal standard from plasma was performed through the use of hexane along with iso-propylalcohol. After liquid-liquid extraction we moved straightforward to the process of the mobile phase in which we had a combination of acetonitrile and potassium dihydrogen phosphate (65:35, v/v). The separation was carried out on a reversed phase C18 column with UV-detector at 210 nm for bupivacaine and 230nm for the internal standard. This particular examination exhibited excellent linearity (r2=0.999) in peak response over the concentration range of 0.1–1.5µg/mL bupivacaine in human plasma. The mean absolute recoveries for 0.5 and 1.5µg/mL of bupivacaine in plasma using the present extraction procedure were 97.43% and 95.03%, respectively. The intra- and inter-day coefficients of variation in the plasma were less than 12% at the lowest, and less than 10% at other concentrations, and the percent error values were less than 6%. The method displayed a high caliber of sensitivity and selectivity for therapeutic monitoring concentrations of bupivacaine in human plasma.