|Thursday, August 28||Analytical Procedures|
A SENSITIVE ANALYSIS OF THE ACTIVE METABOLITE OF BURGODIN® (DESPROPIONYL-BEZITRAMIDE) IN FORENSIC SAMPLES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH FLUORESCENCE DETECTION|
De Baere S., Meyer E., Lambert W., De Leenheer A.
Bezitramide (Burgodin®), a potent, long-acting, orally active narcotic analgesic, is often abused by drug-addicts in Europe. Bezitramide is a prodrug which is readily hydrolyzed in the gastrointestinal tract to its main metabolite, despropionyl-bezitramide. This metabolite has the same pharmacological activity as the parent compound. A method is presented for the quantitative analysis of despropionyl-bezitramide in different biological matrices (blood, urine, stomach contents, bile and liver). Since blood levels were expected to be very low (4.2-8.2 ng/mL) (1) a highly sensitive method is required. The method entails internal standardization (N-methyl-despropionyl-bezitramide) and a liquid-liquid back extraction with hexane: iso-amylalcohol (95:5, v/v), followed by a sensitive and selective reversed phase high performance liquid chromatographic analysis with fluorescence detection (lexc=280 nm, lem=310 nm). Chromatographic separation was achieved on a Hypersil ODS (C18) 5µ column, under isocratic conditions using a 69:31 (v/v) mixture of methanol:acetonitrile (50:50, v/v) and 0.1M ammonium acetate, as the eluent. Under the described conditions, the quantification limit for despropionyl-bezitramide in blood and urine was 1 ng/mL. Calibration graphs were prepared for blood and urine and were linear over the concentration range from 1 to 50 ng/mL. The method was thoroughly validated. Results are given for the analysis of different biological samples from three fatalities that were due to the combined intake of several drugs, including bezitramide. Moreover, the data from the analysis of 17 urine samples, all from known drug abusers, are presented.
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