Key Laboratory of Environmental Medicine Engineering, Ministry of Education, Southeast University, Nanjing 210009, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Nov 1;938:80-5. doi: 10.1016/j.jchromb.2013.08.032. Epub 2013 Sep 5.
A method was established for the determination of three phenolic environmental estrogens, namely bisphenol A (BPA), nonylphenol (NP) and octylphenol (OP), in urine from women of uterine leiomyoma group (n=49) and control group (n=29), by using solid-phase extraction (SPE) coupled with liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Urine samples were spiked with 2,4,6-tribromophenyl-terminated tetrabromobisphenol-A carbonate oligomer (TBBPA) and nonylphenol D8 (NP-D8) as internal standard (I.S.) and de-conjugated by adding β-glucuronidase and sulfatase before the SPE. The extraction recoveries of BPA, NP and OP were more than 73.3%; the standard curve was linear over the validated concentrations in the range of 1.0-100.0ng/mL and the limits of detection (LOD) of BPA, NP and OP were 0.32ng/mL, 0.18ng/mL and 0.15ng/mL, respectively. Moreover, by analysing quality control urine samples in 5 days, the results showed that the method was precise and accurate, for the intra- and inter-day CV% within 15.2%. Except that OP was not found (<LOQ) in any of the control urine samples, the three phenolic environmental estrogens were detected in all urine samples. For the uterine leiomyoma women, the mean concentrations of BPA, NP and OP were 13.9±12.7ng/mL, 2.77±2.22ng/mL and 4.09±5.51ng/mL (mean±SD), respectively. For the control group, the mean concentrations of BPA and NP were 8.50±12.2ng/mL and 3.84±3.90ng/mL (mean±SD), respectively. The Wilcoxon rank sum test was employed for the comparison of BPA and NP between and control in 2 subgroups defined by the number of gravidity (≤3 and >3). NP levels were significantly higher in uterine leiomyoma patients than control group in low gravidity subgroup. Though BPA levels in experimental and control groups were not significantly different, the mean levels and concentration distribution were different. The study suggested that there is certain relationship between exposure concentrations of phenolic environmental estrogens and uterine leiomyoma disease.
建立了固相萃取（SPE）-液相色谱-串联质谱法（HPLC-MS/MS）测定子宫肌瘤组（n=49）和对照组（n=29）妇女尿液中三种酚类环境雌激素（双酚 A（BPA）、壬基酚（NP）和辛基酚（OP））的方法。尿样中加入 2,4,6-三溴苯端四溴双酚 A 碳酸酯低聚物（TBBPA）和壬基酚 D8（NP-D8）作为内标（IS），并用β-葡萄糖醛酸酶和硫酸酯酶处理后进行 SPE。BPA、NP 和 OP 的提取回收率均大于 73.3%；标准曲线在 1.0-100.0ng/mL 的验证浓度范围内呈线性，BPA、NP 和 OP 的检出限（LOD）分别为 0.32ng/mL、0.18ng/mL 和 0.15ng/mL。此外，通过分析 5 天内的质控尿样，结果表明该方法精确准确，日内和日间 CV%均小于 15.2%。除了在任何对照尿样中均未检出 OP（<LOQ）外，所有尿样中均检测到这三种酚类环境雌激素。对于子宫肌瘤女性，BPA、NP 和 OP 的平均浓度分别为 13.9±12.7ng/mL、2.77±2.22ng/mL 和 4.09±5.51ng/mL（平均值±标准差）。对于对照组，BPA 和 NP 的平均浓度分别为 8.50±12.2ng/mL 和 3.84±3.90ng/mL（平均值±标准差）。Wilcoxon 秩和检验用于比较两组（生育次数≤3 和>3）中 BPA 和 NP 的组间差异。在低生育次数亚组中，子宫肌瘤患者的 NP 水平明显高于对照组。尽管实验组和对照组的 BPA 水平没有显著差异，但平均水平和浓度分布不同。该研究表明，酚类环境雌激素的暴露浓度与子宫肌瘤疾病之间存在一定的关系。
