Schlüsener Michael P, Bester Kai
Waste and Wastewater Management, University of Duisburg Essen, Campus Essen, Universitaetsstr. 15, D-45141 Essen, Germany.
Rapid Commun Mass Spectrom. 2005;19(22):3269-78. doi: 10.1002/rcm.2189.
In this study we present a high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) method which has been elaborated to analyse steroid hormones, hormone conjugates, oral contraceptives and macrolide antibiotics unchanged in unfiltered influents and effluents of sewage treatment plants (STPs). HPLC separation of the steroid hormones was achieved in 35 min, as well as those of the antibiotics. The analytes were extracted by solid-phase extraction, followed by clean-up using size exclusion chromatography (SEC). For the final quantification HPLC/MS/MS was used. The two ionisation modes, electrospray ionisation (ESI) and atmospheric pressure chemical ionisation (APCI), in HPLC/MS/MS were compared for the analysis of steroid hormones. For quantitative results drastic matrix effects were observed while using ESI. These effects were less pronounced while using APCI. These pitfalls were additionally reduced by clean-up using SEC as well as isotope dilution. Additionally, two multiple reaction monitoring (MRM) transitions per compound were used to prevent false positive results. Recovery experiments with spiked tap water with concentrations varying from 1 to 1000 ng/L gave constant recovery rates: The recovery rates for the hormones and conjugates ranged from 58 to 107%, those of the contraceptives ranged from 83 to 109%. The relative standard deviation was found to be 7 to 24% and the limits of detection were 0.1 to 4.5 ng/L. The recovery rates of the macrolide antibiotics ranged from 76 to 103%, while the relative standard deviation was found to be 7 to 14% and the limits of detection ranged from 0.6 to 1.8 ng/L. The maximum concentrations found in influents of a STP was 470 ng/L for estriol and 1200 ng/L for erythromycin.
在本研究中，我们提出了一种高效液相色谱/串联质谱（HPLC/MS/MS）方法，该方法经过精心设计，用于分析污水处理厂（STP）未过滤进水和出水中未变化的甾体激素、激素结合物、口服避孕药和大环内酯类抗生素。甾体激素和抗生素在35分钟内实现了HPLC分离。分析物通过固相萃取进行提取，随后使用尺寸排阻色谱（SEC）进行净化。最终定量采用HPLC/MS/MS。比较了HPLC/MS/MS中的两种电离模式，即电喷雾电离（ESI）和大气压化学电离（APCI），用于甾体激素的分析。使用ESI时观察到显著的基质效应。使用APCI时，这些效应不太明显。通过使用SEC净化以及同位素稀释，进一步减少了这些问题。此外，每种化合物使用两个多反应监测（MRM）转换来防止假阳性结果。对添加浓度从1到1000 ng/L的加标自来水进行的回收率实验给出了恒定的回收率：激素和结合物的回收率范围为58%至107%，避孕药的回收率范围为83%至109%。相对标准偏差为7%至24%，检测限为0.1至4.5 ng/L。大环内酯类抗生素的回收率范围为76%至103%，而相对标准偏差为7%至14%，检测限范围为0.6至1.8 ng/L。在一个污水处理厂的进水中发现的最高浓度，雌三醇为470 ng/L，红霉素为1200 ng/L。
