Molecular Recognition Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
Department of Chemistry and Biochemistry, University of California, Los Angeles, 607 Charles E. Young Drive East Box 951569, Los Angeles, CA 90095-1569, USA.
Sci Total Environ. 2014 Feb 1;470-471:1408-13. doi: 10.1016/j.scitotenv.2013.07.037. Epub 2013 Aug 5.
Phthalates are used in industry products, household items, and medical tools as plasticizers. Human exposure to phthalates has raised concern about its toxicity. In the present study, optimization was conducted for the simultaneous analysis of eight kinds of phthalate metabolites using gas chromatography-mass spectrometry (GC-MS): MEP, MiBP, MnBP, MBzP, MiNP, MEHP, MEOHP, and MEHHP. In order to minimize the matrix effect and to do quantitative analysis, isotope dilution and LLE-GC-MS methods were performed. Urine samples were enzymatically hydrolyzed, extracted with a mixture of n-hexane and ethyl ether (8:2; v:v), and subsequently derivatized with trimethylsilylation. All eight kinds of analytes showed clear resolution and high reproducibility in GC-MS results. The method detection limit ranged from 0.05 ng/mL to 0.2 ng/mL. Calibration curves were found to be linear from 0.2 to 100 ng/mL with -(2)>0.992. The relative standard deviation of the intraday precision using water and urine ranged from 2.1% to 16.3%. The analysis was performed with urine samples that were collected from adults residing in the Republic of Korea. The analyzed concentration results were compared according to gender and region. As a result, DEHP metabolites showed the highest detected concentration (75.92 μg/g creatinine, 100%), and MiNP, a metabolite of DiNP, showed the lowest detected concentration (0.42 μg/g creatinine, 22.5%). On average, female urine (200.76 μg/g creatinine) had a higher detected concentration of ∑8 phthalate metabolites than male urine. Samples from rural regions (211.96 μg/g creatinine) had higher levels than samples from urban regions.
邻苯二甲酸酯被广泛应用于工业产品、家庭用品和医疗工具中，作为增塑剂。人类接触邻苯二甲酸酯引起了人们对其毒性的关注。在本研究中，采用气相色谱-质谱联用（GC-MS）对 8 种邻苯二甲酸酯代谢物（MEP、MiBP、MnBP、MBzP、MiNP、MEHP、MEOHP 和 MEHHP）进行了同时分析的优化。为了最小化基质效应并进行定量分析，采用了同位素稀释和液-液萃取-GC-MS 方法。尿液样品经酶解后，用正己烷和乙醚（8:2，v:v）混合液提取，然后用三甲基硅烷化衍生化。在 GC-MS 结果中，所有 8 种分析物均表现出清晰的分辨率和高重现性。方法检测限范围为 0.05-0.2ng/mL。校准曲线在 0.2-100ng/mL 范围内呈线性，相关系数（r）均大于 0.992。采用水和尿液进行日内精密度测定，其相对标准偏差（RSD）在 2.1%-16.3%之间。分析采用了来自韩国成年人的尿液样本。根据性别和地区对分析浓度结果进行了比较。结果表明，DEHP 代谢物的检出浓度最高（75.92μg/g 肌酐，100%），DiNP 代谢物 MiNP 的检出浓度最低（0.42μg/g 肌酐，22.5%）。平均而言，女性尿液（200.76μg/g 肌酐）中∑8 种邻苯二甲酸酯代谢物的检出浓度高于男性尿液。农村地区（211.96μg/g 肌酐）的样本浓度高于城市地区。
