Sabaredzovic Azemira, Sakhi Amrit Kaur, Brantsæter Anne Lise, Thomsen Cathrine
Division of Environmental Medicine, Norwegian Institute of Public Health, P.O. Box 4404, Nydalen, 0403 Oslo, Norway.
Division of Environmental Medicine, Norwegian Institute of Public Health, P.O. Box 4404, Nydalen, 0403 Oslo, Norway.
J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Oct 1;1002:343-52. doi: 10.1016/j.jchromb.2015.08.040. Epub 2015 Aug 30.
Phthalates (dialkyl or alkyl phenyl esters of phthalic acid, benzene-1.2-dicarboxylic acid) are a group of industrial chemicals that have been used for more than 50 years. Phthalates are ubiquitous and can potentially have adverse effects on humans. The present study presents an accurate, sensitive and automated analytical method for measuring 12 phthalate metabolites (free and conjugated) in human urine using on-line solid phase extraction coupled to high performance liquid chromatography - electrospray ionization - tandem mass spectrometry. A small volume of urine sample (300μL) is required. Glucoronidated phthalate metabolites are deconjugated by incubation with glucoronidase enzyme (Escherihia coli-K 12) and the reaction is stopped by adding formic acid. This is the only sample preparation needed prior to injection into the column switching system. Thus, the method involves minimal sample handling and minimizes possible contaminations from the surroundings. The method was validated by spiking synthetic urine at 5-8 levels in the range of 0.1-500ng phthalate metabolites/mL synthetic urine. The method is sensitive with limits of detection in the low nanogram range, and rapid with a total run time about 25min. The accuracy was between 90 and 120 % and the intermediate precision was given as relative standard deviation was below 20% for most of the compounds. The high sensitivity, high throughput and minimal manual handling make the method suitable for large-scale biomonitoring studies. The present method was applied for the determination of phthalate metabolites in urine samples from 116 pregnant women, a subproject within the Norwegian Mother and Child Cohort Study. Concentrations of all the twelve phthalate metabolites was >LOQ in 100% of the samples analysed. Mean urinary concentrations for different phthalate metabolites ranged from 1 to 100ng/mL, the highest concentrations were observed for di-2-ethylhexyl phthalate (DEHP) metabolites and lowest for di-iso-nonyl phthalate (DiNP) metabolites. The urinary concentrations for most of the phthalate metabolites in the present study were found to be in the same range as found in other studies of pregnant women.
邻苯二甲酸酯（邻苯二甲酸的二烷基酯或烷基苯基酯，即苯 - 1,2 - 二甲酸酯）是一类已使用超过50年的工业化学品。邻苯二甲酸酯广泛存在，可能对人体产生不良影响。本研究提出了一种准确、灵敏且自动化的分析方法，用于通过在线固相萃取结合高效液相色谱 - 电喷雾电离 - 串联质谱法测定人尿中12种邻苯二甲酸酯代谢物（游离型和结合型）。所需尿样体积小（300μL）。结合型邻苯二甲酸酯代谢物通过与葡萄糖醛酸酶（大肠杆菌 - K12）孵育进行去结合，反应通过添加甲酸终止。这是注入柱切换系统前唯一需要的样品制备步骤。因此，该方法涉及的样品处理最少，并将来自周围环境的可能污染降至最低。该方法通过在0.1 - 500ng邻苯二甲酸酯代谢物/mL合成尿的范围内以5 - 8个水平向合成尿中加标进行验证。该方法灵敏，检测限在低纳克范围内，且快速，总运行时间约25分钟。准确度在90%至120%之间，对于大多数化合物，中间精密度以相对标准偏差低于20%给出。高灵敏度、高通量和最少的人工操作使该方法适用于大规模生物监测研究。本方法应用于挪威母婴队列研究中的一个子项目，即测定116名孕妇尿样中的邻苯二甲酸酯代谢物。在所有分析的样品中，所有十二种邻苯二甲酸酯代谢物的浓度均高于定量限。不同邻苯二甲酸酯代谢物的尿平均浓度范围为1至100ng/mL，邻苯二甲酸二（2 - 乙基己基）酯（DEHP）代谢物的浓度最高，邻苯二甲酸二异壬酯（DiNP）代谢物的浓度最低。本研究中大多数邻苯二甲酸酯代谢物的尿浓度与其他孕妇研究中的浓度范围相同。
