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Clauwaert KM, Van Bocxlaer JF, De Letter EA, Van Calenbergh S, Lambert WE, De Leenheer AP. 
“Determination of the designer drugs 3, 4-methylenedioxymethamphetamine, 3,4-methylenedioxyethylamphetamine, and 3,4-methylenedioxyamphetamine with HPLC and fluorescence detection in whole blood, serum, vitreous humor, and urine”. 
Clin Chem. 2000 Dec 26;46(12):1968-77.

BACKGROUND: The popular designer drugs 3, 4-methylenedioxymethamphetamine MDMA and 3, 4-methylenedioxyethylamphetamine MDEA can be determined in serum, whole blood, and urine, but also in vitreous humor. The latter matrix is interesting when dealing with decomposed bodies in a toxicological setting.

METHODS: After extraction, chromatographic separation was achieved on a narrow-bore C18 column by gradient elution with fluorometric detection results were confirmed by liquid chromatography-tandem mass spectrometry LC-MS/MS.

RESULTS: The method was linear over the range of 2-1000 microg/L for whole blood, serum, and vitreous humor, and 0.1-5 mg/L for urine. Extraction recoveries were >70, imprecision CV was 2.5-19, and analytical recoveries were 95.5-104.4. The limit of detection LOD and the limit of quantification LOQ were 0.8 and 2 microg/L, respectively, for whole blood, serum, and vitreous humor, and 2.5 microg/L and 0.1 mg/L, respectively, for urine. Excellent correlations between the quantitative LC-fluorescence and LC-MS/MS results were obtained. We found the following concentrations in a thanatochemical distribution study in rabbits: in serum, 5.3-685 microg/L for MDMA and from the LOQ to 14.5 microg/L for 3, 4-methylenedioxyamphetamine MDA in whole blood, 19.7-710 microg/L for MDMA and from the LOQ to 17.8 microg/L for MDA in vitreous humor, 12.1-97.8 microg/L for MDMA and from the LOQ to 3.86 microg/L for MDA. In routine toxicological urine samples, concentrations ranged from LOQ to 14.62 mg/L for MDA, from LOQ to 157 mg/L for MDMA, and from LOQ to 32.54 mg/L for MDEA. CONCLUSIONS: The HPLC method described is sensitive, specific, and suitable for the determination of MDMA, MDEA, and MDA in whole blood, serum, vitreous humor, and urine.
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