Determination of Bupivacaine in Human Plasma by HPLC

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Abstract
A rapid, sensitive, selective and reproducible reversed- phase high-performance liquid chromatographic method has been developed and validatd for the quantification of bupivacaine in human. Bupivacaine and the internal standard, lidocaine, are extracted from alkalinized plasma using n-hexane before chromatographing on a reversed-phase system. The mobile phase comprises 62 parts of 0.05 M phosphate buffer and 38 parts of methanol at pH 5.9 and it is pumped at 1.0 ml/min. The retention times of bupivacaine and lidocaine were observed to be 3.8 and 5.9 min respectively. Ultraviolet detection at 254 nm enabled a limit of detection of 25 ng/ml to be achieved. The reproducibility of the method was good at 1000 ng ml (C. V.=4.3%, n=6). The method is linear from 50 to 3200 ng/ml. The high sensitivity and the speed at which this assay can be performed makes it especially useful for estimating bupivacaine in human plasma.