污水中18种类固醇激素化合物的痕量分析

Analysis of 18 Trace Steroid Hormone Compounds in Wastewater

采用HLB固相萃取柱和超高效液相色谱-串联质谱法(UPLC-MS/MS),在多反应监测(MRM)模式下建立了污水处理厂进出水样中18种类固醇激素(群勃龙、诺龙、雄烯二酮、睾酮、炔诺酮、甲基睾酮、孕酮、康力龙、羟孕酮、泼尼松、氢化泼尼松、六甲强龙、醋酸甲地孕酮、安宫黄体酮、倍他米松、地塞米松、醋酸泼尼松龙和醋酸氢化可的松)的快速测定方法。利用单因素试验和正交试验对固相萃取(SPE)条件进行优化,并优化了UPLC-MS/MS分析条件。18种类固醇激素在1.0～200μg.L-1范围内呈良好线性,相关系数r2均不低于0.997 6,方法的检出限(S/N=3)为1.49～10.59 ng.L-1,在20～100 ng.L-1加标范围内的平均回收率为67%～109%,相对标准偏差(RSD)均不高于13.9%。将该方法用于南京地区4个综合污水处理厂进出水样中类固醇激素的测定,发现4个污水处理厂进水中均含有诺龙、雄烯二酮、睾酮、羟孕酮、泼尼松和氢化泼尼松,其中泼尼松含量最高,出水中也有一定量的类固醇激素。

A sensitive ultra-performance liquid chromatography -electrospray tandem mass spectrometric method（ UPLC - MS/MS） combined with solid-phase extraction（SPE） was established for the determination of eighteen steroid hormones, including prednisolone, prednisone, methylprednisolohe, hetamethasone, dexamethasone, trenbolone, prednisolone acetate, hydrocortisone acetate, nortestosterone, testosterone, stanozolol, norethindrone, hydroxyprogesterone, methyltestosterone, androstenedione, megestrol acetate, progesterone and medroxyprogesterone acetate in the influent and final effluent from wastewater treatment plant（WWTP）. The operation conditions influencing the SPE performance were optimized by single factor experiments and orthogonal test, and UPLC - MS/ MS experimental parameters were also studied in detail. The results indicated that, under the optimal conditions, the calibration curves of eighteen steroid hormones were linear in the range of 1.0 -200μg·L^-1 with their correlation coefficients （ r2 ） not less than 0. 997 6. The detection limits for the method ranged from 1.49 ng·L^-1 to 10.59 ng ·L^-1. The recoveries of WWTP water samples at spiked levels of 20 - 100 ng ·L^-1were between 67% and 109% with RSDs not more than 13.9%. The method was applied in the determination of steroid hormones in wastewater from four sewage treat- ment plants in Nanjing. Nortestosterone, androstenedione, testosterone, hydroxyprogesterone, prednisone and prednisolone were found in the influent of the four sewage treatment plants, in which the concentration of prednisone was the highest. A certain amount of steroid hormones were also found in the final effluent.