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Development of a Method for the Quantitation of Benzphetamine Metabolites in Human Urine by High-Performance Liquid Chromatography

Development of a Method for the Quantitation of Benzphetamine Metabolites in Human Urine by High-Performance Liquid Chromatography

A Fujinami, T Miyazawa, Y Kobayashi

Ann Clin Biochem. 1998 Nov;35 ( Pt 6):775-9.

PMID: 9838992

Abstract:

We developed a high-performance liquid chromatography (HPLC) method for quantitating p-hydroxy-N-benzylamphetamine glucuronide (pHBAG) and p-hydroxy-benzphetamine glucuronide (pHBZG), which are urinary metabolites of benzphetamine, in humans. Urine samples were hydrolysed with beta-glucuronidase (EC 3.2.1.31) at 37 degrees C overnight and the treated urine was applied to a solid phase extraction column. After washing the column with water, 0.01 mol/L acetic acid and methanol, pHBA and pHBZ were eluted with dichloromethane:isopropanol:28% ammonium hydroxide (78.4:19.6:2.0 v/v). The eluate was evaporated and the residue was dissolved in acetonitrile: 5 mmol/L 1-pentane sulphonic acid (5:95 v/v) and analysed by HPLC with gradient elution. The amounts of urinary pHBAG and pHBZG excreted by two human subjects after oral administration of 10 mg benzphetamine hydrochloride were determined. About 10-15% of benzphetamine was found to be excreted as pHBAG and pHBZG, and almost all of these metabolites were excreted within 24 h. Urine samples should be collected as early as possible after ingestion of benzphetamine to detect pHBAG and pHBZG.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP5411223	Benzphetamine hydrochloride		5411-22-3	Price

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