Determination of benzene metabolites in urine of mice by solid-phase extraction and high-performance liquid chromatography.
A method was developed for quantitative measurement of trans,trans-muconic acid, catechol, hydroquinone and phenol in urine. Hydrolysis of esterified and glucuronized phenolic compounds was effected by specific enzymes. The hydrolysed mixture was purified and separated by solid-phase extraction with an anion exchanger, followed by extraction with diethyl ether. By using a clean-up procedure the natural background from mouse urine could be reduced, so that the detection limit of the metabolites was in the range 3-60 mg/l. Optimization of the chromatographic conditions resulted in a short high-performance liquid chromatography analysis time. Phenol had the longest retention time of about 10 min. The clean-up procedure could also be used for phenylmercapturic acid, an additional benzene metabolite, but for sensitive high-performance liquid chromatographic detection of phenylmercapturic acid other conditions are necessary.
Schad
H
H
Institut für Arbeits- und Sozialmedizin, Universität Ulm, Germany.
Schäfer
F
F
Weber
L
L
Seidel
H J
HJ
D000818:Animals; D001554:Benzene / Q000378:metabolism*; D002396:Catechols / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006873:Hydroquinones / Q000652:urine*; D051379:Mice; D019800:Phenol; D010636:Phenols / Q000652:urine*; D013011:Sorbic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D002396:Catechols; D006873:Hydroquinones; D010636:Phenols; D019800:Phenol; C005939:muconic acid; D001554:Benzene; C034221:catechol; D013011:Sorbic Acid; C031927:hydroquinone