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    固相萃取结合超高效液相色谱-串联质谱法测定土壤中有机磷酸酯代谢物的含量

    Determination of Organophosphate Ester Metabolites in Soil by Solid Phase Extraction Combined with Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

    • 摘要: 取0.2 g预处理过的土壤样品,加入100 μL 0.1 mg·L−1替代物(磷酸二苯酯-d10)标准溶液,加入2 mL甲醇,超声提取10 min,离心10 min,重复提取1次。合并的上清液经氮吹浓缩至1 mL后,用水稀释至10 mL,并用乙酸钠缓冲溶液调节pH为5,再经Oasis WAX固相萃取柱预先用4 mL含5%(体积分数,下同)氨水的甲醇溶液、4 mL甲醇和4 mL水活化净化,用4 mL含5%氨水的甲醇溶液洗脱。收集洗脱液,于室温(约25 ℃)氮吹浓缩至干,加入500 μL甲醇复溶。分取200 μL复溶液,加入200 μL 0.02 mg·L−1同位素内标磷酸二(1-氯-2-丙基)酯-d12溶液,过0.22 μm滤膜,采用超高效液相色谱-串联质谱法测定其中有机磷酸酯代谢物(mOPEs)的含量,以Acquity UPLC BEH C8色谱柱分离,以不同体积比的甲醇和5 mmol·L−1乙酸铵溶液的混合溶液为流动相进行梯度洗脱,采用电喷雾离子源负离子模式电离,多反应监测模式检测,内标法定量。结果表明,12种mOPEs的质量浓度在0.5~50 μg·L−1内和目标物与同位素内标的峰面积之比呈线性关系,检出限为0.009~0.100 ng·g−1。按照标准加入法进行回收试验,回收率为83.6%~123%,测定值的相对标准偏差(n=6)为0.70%~11%。方法用于实际农田土壤样品的分析,结果显示12种mOPEs在土壤样品中均有不同程度的检出,检出率在50%以上的目标物有6种,总质量分数为0.660~30.4 ng·g−1,平均值为9.01 ng·g−1

       

      Abstract: The pretreated soil sample (0.2 g) was taken, and 100 μL of 0.1 mg·L−1 surrogate (diphenyl phosphate-d10) standard solution was added. The mixture was then ultrasonically extracted for 10 min after addition of 2 mL of methanol, followed by centrifugation for 10 min. This extraction process was repeated once. The combined supernatant was then concentrated to 1 mL by nitrogen-blowing, and diluted to 10 mL with water, and pH was adjusted to 5 with sodium acetate buffer solution. The extract was purified using Oasis WAX solid phase extraction column, which was preconditioned with 4 mL of methanol solution containing 5% (volume fraction, the same below) aqueous ammonia, 4 mL of methanol, and 4 mL of water. The target compounds were eluted with 4 mL of methanol solution containing 5% ammonia aqueous, and the eluate was concentrated to dryness by nitrogen-blowing at room temperature (approximately 25 ℃). The residue was reconstituted in 500 μL of methanol. An aliquot (200 μL) of the reconstituted solution was mixed with 200 μL of 0.02 mg·L−1 isotope internal standard bis (1-chloro-2-propyl) phosphate-d12 solution, and passed through a 0.22 μm filter membrane. The 12 organophosphate ester metabolites (mOPEs) in the filtrate were analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry. The separation was performed on an Acquity UPLC BEH C8 column using a mixed solution of methanol and 5 mmol·L−1 ammonium acetate solution at different volume ratios as the mobile phase for gradient elution. Detection was carried out in the negative mode of electrospray ion source and multiple reaction monitoring mode, and quantification was carried out by the internal standard method. As shown by the results, linear relationships between the peak area ratios of the 12 mOPEs to the isotope internal standard and mass concentrations of the targets were kept in the range of 0.5–50 μg·L−1, with detection limits in the range of 0.009–0.100 μg·g−1. Test for recovery was made by the standard addition method, giving results in the range of 83.6%–123%, with RSDs (n=6) of the determined values in the range of 0.70%–11%. This method was applied to analysis of actual farmland soil samples, and it was shown by the results that 12 mOPEs were all detected to varying degrees in the soil samples, with detection ratio greater than 50% for 6 mOPEs, and total mass fractions in the range of 0.660–30.4 ng·g−1 and the average value of 9.01 ng·g−1.

       

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