Teass A W, DeBord D G, Brown K K, Cheever K L, Stettler L E, Savage R E, Weigel W W, Dankovic D, Ward E
Division of Biomedical and Behavioral Science, Centers for Disease Control and Prevention, Cincinnati, OH 45226.
Int Arch Occup Environ Health. 1993;65(1 Suppl):S115-8. doi: 10.1007/BF00381320.
Epidemiological evidence that occupational exposure to o-toluidine and aniline is associated with an increased risk of bladder cancer led to efforts to identify biomarkers of workplace exposures to these aromatic amines. For the determination of o-toluidine and aniline in worker urine specimens, a method using high performance liquid chromatography (HPLC) followed by electrochemical detection was developed. The limits of detection were 0.6 microgram/l and 1.4 micrograms/l for o-toluidine and aniline, respectively. Recovery of o-toluidine and aniline from spiked urine averaged 86% and 93%, respectively, over a range of 4-100 micrograms/l. Reproducibility in the range 2-100 micrograms/l for analyses of split field samples was 13% (average RSD) for o-toluidine and 16% (average RSD) for aniline. Application of this method to pre- and post-shift samples collected from potentially exposed and unexposed workers indicated elevated concentrations of o-toluidine and aniline in urine from exposed workers. To develop methods for biomarkers of internal dose, o-toluidine binding to the blood proteins hemoglobin and albumin was investigated utilizing in-vivo (rodent) and in-vitro (hemoglobin and albumin) studies. Base-hydrolyzable protein adducts were analyzed by HPLC (fluorescence) and/or GC/electron capture (EC). The methods were compared for sample preparation requirements, selectivity and sensitivity. While the GC/EC method was more sensitive than HPLC, the presence of interfering peaks limited the utility of this approach. Results from these studies suggested that the HPLC method could be useful for determination of o-toluidine exposures in individuals acutely or chronically exposed to high levels.
流行病学证据表明，职业性接触邻甲苯胺和苯胺会增加患膀胱癌的风险，这促使人们努力寻找这些芳香胺在工作场所接触的生物标志物。为了测定工人尿液样本中的邻甲苯胺和苯胺，开发了一种采用高效液相色谱（HPLC）并结合电化学检测的方法。邻甲苯胺和苯胺的检测限分别为0.6微克/升和1.4微克/升。在4 - 100微克/升的范围内，加标尿液中邻甲苯胺和苯胺的回收率平均分别为86%和93%。对于分割样本分析，在2 - 100微克/升范围内，邻甲苯胺的重现性为13%（平均相对标准偏差），苯胺为16%（平均相对标准偏差）。将该方法应用于从可能接触和未接触的工人中采集的班前和班后样本，结果表明接触工人尿液中邻甲苯胺和苯胺的浓度升高。为了开发内部剂量生物标志物的方法，利用体内（啮齿动物）和体外（血红蛋白和白蛋白）研究，对邻甲苯胺与血液蛋白血红蛋白和白蛋白的结合情况进行了研究。通过HPLC（荧光）和/或GC/电子捕获（EC）分析碱可水解蛋白加合物。对这些方法在样品制备要求、选择性和灵敏度方面进行了比较。虽然GC/EC方法比HPLC更灵敏，但干扰峰的存在限制了该方法的实用性。这些研究结果表明，HPLC方法可用于测定急性或慢性高剂量接触邻甲苯胺的个体的接触情况。
