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Determination of Cocaine and Norcocaine in Plasma and Cell Cultures Using High-Performance Liquid Chromatography

Determination of Cocaine and Norcocaine in Plasma and Cell Cultures Using High-Performance Liquid Chromatography

P Bouis, G Taccard, U A Boelsterli

J Chromatogr. 1990 Apr 6;526(2):447-59.

PMID: 2361984

Abstract:

A new simple high-performance liquid chromatographic (HPLC) method was developed for the determination of cocaine and norcocaine. Cocaine and norcocaine in biological samples were buffered to pH 9.0, extracted with diethyl ether and reextracted in a 0.1% aqueous solution of tetramethylammonium hydrogen sulfate (TMAHS) with a theoretical yield of extraction of 100%. The HPLC elution of cocaine and norcocaine was performed using a Spherisorb RP-18, 100 mm x 4.6 mm I.D., 5 microns particle size column with a mobile phase containing acetonitrile-0.1% TMAHS aqueous solution (60:40). The compounds were entirely separated, and a reliable limit of quantitation was set at 20 ng/ml when extracted from 0.5 ml of plasma. No interference with 26 other drugs was found. Cocaine and norcocaine stability studies showed that their half-lives in human plasma incubated at 37 degrees C were 50.8 and 43.2 min, respectively. In contrast, plasma from dogs or rats exhibited only weak or no enzymatic esterase activity towards cocaine and norcocaine resulting in less rapid degradation. Hydrolysis could be efficiently inhibited with sodium fluoride and prevented by storage of the sample at -20 degrees C. The highly sensitive assay also allowed the assessment of the oxidative metabolism pathway of cocaine to norcocaine in primary rat hepatocyte cultures.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP61585226	Norcocaine hydrochloride solution		61585-22-6	Price

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