固相萃取-超高效液相色谱-串联质谱法同时测定水中61种激素

Simultaneous determination of 61 hormones in water by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry

甾体激素类的污染及危害问题不容忽视,为了实现更加全面、准确的高通量分析,研究建立了固相萃取-超高效液相色谱-串联质谱同时测定水中糖皮质激素类(48种)、盐皮质激素类(1种)、雄激素类(4种)及孕激素类(8种)等共61种激素成分的多残留分析方法。采用HC-C18固相萃取柱对大体积(1 L)水样中的目标化合物进行富集净化,乙腈洗脱,以BEH C18色谱柱分离,以01%甲酸水溶液和乙腈为流动相梯度洗脱,用超高效液相色谱-串联质谱仪分离和检测。质谱采用电喷雾正离子电离、多反应监测模式,外标法定量。61种激素在相应的范围内,线性关系良好,相关系数均大于099,方法检出限为005~150 ng/L,在低、中、高3个加标水平下的回收率为623%~1252%,相对标准偏差为11%~105%。将建立的方法应用于太湖流域的地表水、相关区域的地下水以及末梢水的分析,部分地表水和地下水中检出了可的松、丙酸氟替卡松、环索奈德、倍他米松双丙酸酯、氯倍他松丁酸酯、戊酸双氟可龙、卤倍他索丙酸酯、异氟泼尼龙、二氟孕甾丁酯和己酸羟孕酮等10种成分,其余51种成分未检出。该方法操作简单,灵敏度高,准确度好,对后续水安全的监测以及溯源调查有重要意义。对水中激素水平进行了地区分析,提出了未来污水处理工艺应将激素残留作为目标物进行针对性处理的建议。

Concerns over the emergence of steroid hormones as pollutants in water have grown.Steroid hormone compounds present challenges in the simultaneous detection of total residual hormones owing to their analogous structures and diverse types.In this study,we es-tablished a rapid and high-throughput continuous online method based on solid phase extraction(SPE)coupled with ultra performance liquid chromatography-tandem mass spectrometry(UP-LC-MS/MS)for the simultaneous determination of 61 hormone components,including 48 glu-cocorticoids,1 mineralocorticoid,4 androgens,and 8 progesterones,in water.Various SPE columns were investigated to assess their extraction efficiency for enriching and purifying target compounds in a large sample volume(1 L).An HC-C18 SPE column was select-ed because of its superior performance.Acetonitrile was used as a washing solution during SPE to ensure that the majority of the tested substances achieved recoveries exceeding 70%and effectively avoid interferences from water-soluble components.Various C8 and C18 columns were tested,and the optimal HPLC conditions for hormone re-tention were established.We systematically evaluated different UPLC columns and mobile pha-ses,including methanol-water and acetonitrile-water systems with 01%formic acid added to the aqueous phase.The optimized UPLC conditions were as follows:BEH C18 column(100 mm×21 mm,17μm);column temperature,40℃;flow rate,03 mL/min;injection volume,5μL;mobile phase A:01%formic acid aqueous phase;mobile phase B:acetonitrile.Gradient elution was performed as follows:0-05 min,30%B;05-150 min,30%B-75%B;150-180 min,75%B-98%B;180-190 min,98%B;190-191 min,98%B-30%B;191-200 min,30%B.Both positive-and negative-ion modes were explored in the UPLC-MS/MS experiment to ob-tain the full scan of the parent ions,and positive mode was finally selected for electrospray ion-ization(ESI).Two product ions exhibiting strong signals and minimal interference were select-ed for quantitative and qualitative ion analyses,using an external standard method for quantifi-cation.MS/MS was performed in positive-ion(ESI+)mode with multiple reaction monitoring(MRM)scanning.The MS/MS parameters were as follows:atomizing gas pressure,379 kPa;curtain air pressure,241 kPa;spray voltage,5500 V;desolvation temperature,550℃;colli-sion exit voltage(CXP),13 V;intake voltage(EP),10 V;and residence time of each ion pair,05 ms.Other instrument settings,such as the collision energy and declustering voltage,were also optimized.The 61 hormones exhibited excellent linear relationships within their corresponding concen-tration ranges,with correlation coefficients greater than 099.The method detection limits(MDLs)were in the range of 005-150 ng/L.The average recoveries of the 61 hormones across three spiked levels ranged from 623%to 1252%,with relative standard deviations(RSDs,n=6)of 11%-105%.Using this method,we successfully detected 10 hormone components(cortisone,fluticasone propionate,ciclesonide,betamethasone dipropionate,clobetasone butyrate,diflucortolone va-lerate,halobetasol propionate,isoflupredone,difluprednate,and hydroxyprogesterone ca-proate)in various surface water and groundwater samples collected from the Taihu Basin region.The SPE-UPLC-MS/MS method presented in this paper is simple,highly sensitivity,and ex-ceptionally accurate.Thus,it exhibits promising potential for tracing targeted hormone residues in water and will be of great value in monitoring and ensuring water safety.Finally,a regional analysis was conducted on the hormone levels in water,and suggestions were made for the tar-geted treatment of hormone residues in future sewage treatment processes.