Sensitive and specific high-performance liquid chromatographic assay with ultraviolet detection for the determination of cocaine and its metabolites in rat plasma

Journal of Chromatography. B, Biomedical Sciences and Applications
W Pan, M A Hedaya

Abstract

A sensitive, specific and precise HPLC-UV assay was developed to quantitate cocaine (COC) and its metabolites benzoylecgonine (BE), norcocaine (NC) and cocaethylene (CE) in rat plasma. After adding 50 microl of the internal standard solution (bupivacaine, 8 microg/ml) and 500 microl of Sørensen's buffer (pH 6) to 100 microl of rat plasma sample, the mixture was extracted with 10 ml of chloroform. The organic layer was transferred to a clean test tube and was evaporated under nitrogen. The residue was reconstituted in 100 microl of mobile phase and 35 microl was injected onto the HPLC column. The mobile phase consisted of methanol-acetonitrile-50 mM monobasic ammonium phosphate (5:7:63, v/v/v) and was maintained at a flow-rate of 0.4 ml/min. Separation of COC and its metabolites was achieved using a Supelcosil ABZ+plus deactivated reversed-phase column (250x2.1 mm I.D., 5 microm). Calibration curves were linear over the range of 25-5000 ng/ml for COC and its three metabolites. The absolute extraction efficiencies for BE, COC, NC, CE and bupivacaine were 56.6%, 78.6%, 61.1%, 76.4% and 67.0%, respectively. COC and its metabolites were stable in mobile phase for 24 h at room temperature and in rat plasma for 2 weeks at -20degrees C...Continue Reading

References

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Aug 1, 1997·Pharmaceutical Research·M A Hedaya, W J Pan

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Citations

May 22, 1998·Journal of Pharmacological and Toxicological Methods·W J Pan, M A Hedaya
May 8, 2003·Analytical and Bioanalytical Chemistry·B CabovskaA M Stalcup
Jul 20, 2002·The Journal of Pharmacology and Experimental Therapeutics·Hong SunStephen Brimijoin
Apr 3, 1999·Journal of Pharmaceutical Sciences·W J Pan, M A Hedaya

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