Hair analysis for fenfluramine and norfenfluramine as biomarkers for N-nitrosofenfluramine ingestion.
In this paper, a high performance liquid chromatographic method with fluorescence detection (HPLC-FL) for the determination of fenfluramine (Fen) and norfenfluramine (Norf) in human hair as biomarker metabolites of N-nitrosofenfluramine (N-Fen) is described. Washed and cut hair segments were extracted by ultrasonication for 1h at room temperature in methanol. The extract was evaporated and applied for derivatization with the fluorescent reagent 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl). An HPLC-FL analysis was performed using an ODS column with mobile phase composition of acetonitrile and water (65:35, v/v) and monitored at 430 nm (excitation 325 nm). The method was sensitive with detection limits of 36 and 16 pg/mg hair for Fen and Norf, respectively. The linearity was assessed in the range 0.036-144 ng/mg for Fen and 0.016-127 ng/mg for Norf with correlation coefficients larger than 0.999. The method was successfully used for the segmental determination of Fen and Norf in hair samples obtained from hospitalized patients diagnosed with hepatotoxicity and suspected to ingest N-Fen. Both Fen and Norf could be detected in these patients' hair samples in the ranges 43-1389 pg/mg for Fen and 18-680 pg/mg for Norf and the results showed that the patients might ingest N-Fen for a period of not less than 5 months. As well, the method was applied for the determination of Fen and Norf in rats that possess pigmented and non-pigmented hair after an intraperitoneal administration of Fen. Both compounds were determined in black as well as in white hair.
Division of Analytical Research for Pharmacoinformatics, Department of Clinical Pharmacy, Course of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Amal
A
Kaddoumi
Mitsuhiro
M
Wada
Mihoko N
MN
Nakashima
Kenichiro
K
Nakashima
D000818:Animals; D001067:Appetite Depressants / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry; D015415:Biomarkers / Q000032:analysis; D056486:Chemical and Drug Induced Liver Injury; D002851:Chromatography, High Pressure Liquid; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry; D005277:Fenfluramine / Q000008:administration & dosage / Q000009:adverse effects / Q000031:analogs & derivatives* / Q000032:analysis*; D005554:Forensic Medicine; D006197:Hair / Q000737:chemistry*; D006801:Humans; D015394:Molecular Structure; D009642:Norfenfluramine / Q000032:analysis*; D051381:Rats; D011924:Rats, Zucker; D015203:Reproducibility of Results; D017367:Selective Serotonin Reuptake Inhibitors / Q000008:administration & dosage / Q000009:adverse effects / Q000032:analysis*; D013050:Spectrometry, Fluorescence
D001067:Appetite Depressants; D015415:Biomarkers; D004365:Drugs, Chinese Herbal; D005277:Fenfluramine; D009642:Norfenfluramine; D017367:Selective Serotonin Reuptake Inhibitors; C493137:nitrosofenfluramine