Determinação do 1-hidroxipireno em amostras de urina por cromnatografia líquida de alta eficiência - Estudo dos parâmetros de validação

Abstract

Polycyclic aromatic hydrocarbons (PAHs) constitute a group of chemical substances formed during thermal degradation of organic materials released in the environment. At present, several hundreds of papers reporting adverse effects produced by PAHs are mainly related to cancer development. According to International Agency for Research on Cancer (IARC), six PAHs are probably human carcinogenic. Considering the toxicological properties and with the aim to prevent occupational exposures to these substances, a biological monitoring of urinary 1-hydroxypyrene is recommended. The first step to achieve this goal is to validate a methodologyfor urinary 1-OHP quantification. This work optimized the conditions of a high performance liquid chromatography system (HPLC) with fluorescence detection and validated the analytical parameters for urine 1-OHP determinations. The method includes an enzymatic hydrolysis (b-glucuronidase/ arylsulfatase) followed by a solid phase extraction - SPE (C18). In addition a chromatographic separation was accomplished using a Supelcosil™ LC-PAH (15 cm × 4.6 mmID × 5 μm) with a gradient phase (methanol 40 and 100%) and finally a selective detection (excitation - 242 nm and emission - 388 nm). The method showed linearity (r2= 0.999) with the studied range of concentration (0.2 to 40.0 ng/mL). Detection and quantification limit were 0.5 ng/mL and 1.0 ng/mL, respectively. Relative standard deviation of the method was good (CV<15%), as also was the reproducibility (recovery maintained between 80 and 120%). In conclusion, this validated method can be considered useful for the urinary 1-OHP analyses.