Method development of di-(2-ethylhexyl) phthalate metabolites detection by dispersive liquid–liquid microextraction gas chromatography–mass spectrometry from urine

Aims: Phthalates (e.g., diethylhexyl phthalate) act as one of the endocrine disruptive compounds and can have adverse effects on different biological pathways including hormonal receptors, affecting the synthesis, secretion, or metabolism of hormones. The aim of this study was, method development of the dispersive liquid-liquid microextraction (DLLME), to determine the monoesters of diethylhexyl phthalate in children and adolescents in Isfahan city, Iran. Materials and Methods: This study was conducted in 2016 (during a year) on 242 children and adolescents, aged 6–18 years, living in Isfahan, Iran. In this method, acetonitrile and chlorobenzene were used as disperser and extractant, respectively, to extract the investigated monoesters. The analytes were determined by gas chromatography–mass spectrometry. Results: Acetonitrile (as disperser) and chlorobenzene (as extractant) were used for monoester phthalates extraction (750 μl of acetonitrile and 80 μl of chlorobenzene). The results indicated that the recovery and relative standard deviation (RSD) of the utilized method were 55–109 and 6.3%–13.2%, respectively. The limit of detection and limit of quantification (LOQ) of the method were 0.024–0.088 and 0.05–0.48 μg/L, respectively. Monoethylhexyl phthalate (MEHP), monoethyl oxo-hexyl-phthalate (MEOHP), and monoethyl hydroxyl-hexyl-phthalate (MEHHP) were observed in 99.6%, 95.87%, and 96.28% of the studied samples, respectively. The mean concentration of MEHP, MEOHP, and MEHHP in the study population was 151.7 ± 143.8, 258.26 ± 143.07, and 194.17 ± 147.3 μg/L, respectively. Conclusion: Acetonitrile (as disperser) and chlorobenzene (as extractant) were suitable solvents for phthalate monoesters extraction in method development of DLLME. On the other hand, the mean concentrations were very high values in comparison with other similar studies that require the attention and legislation regarding limitation of the application of phthalate compounds.