Direct analysis of phenol, catechol and hydroquinone in human urine by coupled-column HPLC with fluorimetric detection

G. Marrubini, E. Calleri, T. Coccini, A. F. Castoldi, L. Manzo

Research output: Contribution to journalArticle


Phenol, catechol, and hydroquinone, are urinary end-products of the metabolism of benzene, nutrients, drugs, and endogenous substances. Recent research demonstrated that phenol, catechol, and hydroquinone, may have themselves a role in the carcinogenicity of benzene and in mechanisms that lead to leukemia. In this respect there is the need of rapid, low-cost, and possibly direct methods to quantitate these phenolic metabolites. Three single-residue coupled-column HPLC methods with fluorimetric detection (LC-LC-FLD) are described for the direct quantitation of phenol, catechol, and hydroquinone, in human urine. After enzymatic hydrolysis of the corresponding beta-glucuronoconjugates and sulfates, urine was directly injected into the LC-LC analyzer. The LC-LC-FLD procedure allowed base-to-base separation of the target compounds from urine interferents and good linearity (r2 = 0.998) within the ranges studied (0.5-50 mg L-1 for phenol, 0.35-35 mg L-1 for catechol and 0.2-10 mg L-1 for hydroquinone). Despite the high background levels of these metabolites in human urine, within- and inter-session precision expressed as RSD% was better than 20% on spiked and on authentic urine samples obtained from benzene-exposed workers. Accuracy expressed as the recovery ratio between measured and nominal concentration in spiked urine was comprised between 93% and 115% for the three metabolites. The column switching system was fully automated and computer-controlled, and was applied to the determination of phenol, catechol, and hydroquinone in urine samples showing a sample throughput of at least 20-30 samples per day.

Original languageEnglish
Pages (from-to)25-31
Number of pages7
Issue number1-2
Publication statusPublished - Jul 2005


  • Column liquid chromatography
  • Coupled-columns
  • Human urine analysis
  • Phenols and hydroquinone

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Analytical Chemistry

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