3,3'-dichlorobenzidine (DCB) is suspected to be carcinogenic in experimental animal and human. Several studies have investigated excretion of metabolites in urine, hemoglobin adduction and cancer incidence among workers exposed occupationally to DCB. In these researches, metabolites of DCB had a very important role. The purpose of this study was to develop the urinary monitoring method of its metabolites from rats exposed with DCB, by easily synthesizing them in the laboratory. N,N'-diacetyl-DCB was easily synthesized with DCB in pyridine by adding sufficient acetyl chloride or acetic anhydride. N-acetyl-DCB was isolated from the supernatant, which made by adding 21µl acetyl chloride (more 3 times than DCB in moles) to 32 mg DCB in 2 ml pyridine and 0.3 ml acetic acid. Gas chromatography/mass spectrometry (GC/MS) was used for the identification, gas chromatography nitrogen phosphorous detection (GC-NPD) for the quantification and gas chromatography flame ionization detection (GC-FID) for the determination of purity. The base peak of DCB, N-acetyl-DCB and N,N'-diacetyl-DCB was 252 m/z. The other main peaks were 294 m/z for N-acetyl-DCB, and 294 and 336 m/z for N,N'-diacetyl-DCB. The purities of N-acetyl-DCB and N,N'-diacetyl-DCB were identified as 98.82 and 98.72% by GC-FID, respectively. After treatment orally to rats with 20mg DCB/kg body weight for 2 weeks, the urinary excretion amount of DCB was nearly constant at range of 0.11-0.18 mg/L for 2 weeks. But excretion of N-acetyl-DCB was continually increased from 1.30 mg/L on 1st day to 4.15 mg/L on 14th day. And level of N,N'-diacetyl-DCB in urine was sharply increased from 2.13 mg/L on 1st day to 11.00 mg/L on 14th day.
Parathion is an organophosphate pesticide being legally applied for the purpose of agriculture and is being manufactured in Korea. A gas chromatography/mass spectrometric method was developed for the determination of parathion urinary metabolite, p-nitrophenol. p-Nitrophenol was extracted from weak acidic urine, and then measured by gas chromatography-mass spectrometry (selected ion monitoring). The recovery of pnitrophenol in the overall procedure was 88.2%. The detection limit of the assay was 1.0 μg/L based upon assayed urine of 2.0 mL. The method was applied to the determination of p-nitrophenol in urine of workers of a parathion industry. Spot urines of workers of a parathion industry were sampled at the end of shift and pnitrophenol was analyzed using above developed method. p-Nitrophenol could be detected in all of the urine samples at concentrations varying from 3.0 to 681 μg/L.
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