Determination of harmane and harmine in human blood using reversed-phased high-performance liquid chromatography and fluorescence detection.
A number of tremorogenic beta-carboline alkaloids have been found in common plant-derived foodstuffs, beverages, and inhaled substances. Because of their natural presence in the food chain, there is a growing concern regarding the potential risks of certain essential tremors associated with the long-term, low-level dietary exposure to these alkaloids. The purpose of this study was to develop an effective analytical method to determine blood levels of two major beta-carboline derivatives, harmane and harmine. Human blood was extracted with ethyl acetate and methyl-t-butyl ether (2:98) under an alkaline condition. After evaporation of organic solvent, the samples were reconstructed in methanol. The samples were fractionated on a 250 x 4.6-mm C18 reversed-phase column with an isocratic mobile system consisting of 17.5 mM potassium phosphate buffer (ph 6.5) and methanol (30:70), followed by an on-line fluorescence detection. The method had the detection limit to determine 206 and 81 pg/ml of harmane and harmine, respectively, in 10 ml of human blood. The intraday precision (C.V.) at 25 ng/ml was less than 6.7 and 3.4% for harmane and harmine, respectively. The interday precision was 7.3% for harmane and 5.4% for harmine. The method has proven sensitive, reproducible, and thus useful for both laboratory and clinical studies of beta-carboline toxicities.
Zheng
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W
Division of Environmental Health Sciences, School of Public Health, Columbia University, New York, New York, 10032, USA. wz18@columbia.edu
Wang
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Barnes
L F
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Guan
Y
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Louis
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ED
D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods* / Q000706:statistics & numerical data; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D006247:Harmine / Q000031:analogs & derivatives* / Q000097:blood*; D006801:Humans; D051381:Rats; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D006247:Harmine; C005010:harman