【摘要】 目的建立超高效液相色谱-串联质谱法测定甘蔗、甘蔗渣、呕吐物、血清和尿液中3-硝基丙酸(3-NPA)含量的方法。方法样品加入乙腈和氯化钠超声振荡提取并离心,上清液经PSA固相萃取柱净化后用10%氨水-甲醇溶液(10∶90,V/V)洗脱,洗脱液用氮气吹至近干,用0.4%甲酸水溶解,0.22μm微孔滤膜过滤。选用Waters acquity BEH C18色谱柱(2.1 mm×150 mm×1.7μm)、乙腈-水为流动相、梯度洗脱分离、ESI源、负离子模式、多反应监测方式,基质匹配外标法定量测定3-NPA的含量。结果在1.0~50μg/kg的浓度范围内,3-NPA在甘蔗和血清等基质中有良好的线性关系,相关系数分别为0.9993和0.9998,方法检出限为0.06~0.30μg/kg,定量限为0.20~1.0μg/kg;在加标水平为1.0、10.0和100.0μg/kg时3-NPA的加标回收率在86.9%~102.0%之间,相对标准偏差在1.80%~4.19%之间。结论超高效液相色谱-串联质谱法可准确、快速测定各种中毒样品中的3-NPA,可用于因误食霉变甘蔗引起中毒的应急事件处理。更多还原

【Abstract】 Objective An ultra-performance liquid chromatography-tandem mass spectrometric method was established for determination of 3-nitropropionic acid of sugarcane,sugarcane bagasse,vomit,serum and urine. Methods The 3-nitropropionic acid in poisoning samples was extracted by acetonitrile in super-sonic instrument. The supernatant was cleaned up with PSA column and eluted with 10% ammonia watermethanol( 10∶ 90,V / V),then the purified solution was concentrated by nitrogen,dissolved with water( containing 0. 4% formic acid) and cleaned by 0. 22 μm millipore filter. The filtrate was detected by ultra-performance liquid chromatography-tandem mass spectrometry,identified by electrospray ionization( ESI) in negative mode using multiple reaction monitoring,and quantified with external standards of sample matrix matching.The sample extract was separated on an acquity BEH C18column( 2. 1 mm × 150 mm ×1. 7 μm) by gradient elution in 10 minutes with acetronitrile-water as mobile phase.Results The calibration curves of 3-nitropropionic acid residues showed good linearity in the range of 1. 0- 50 μg / kg with correlation coefficient of 0. 9993 or 0. 9998. Thedetection limits of the method were from 0. 06 μg / kg to 0. 30 μg / kg,and limits of quantitation ranged from 0. 20 μg / kg to 1. 0 μg / kg. The recoveries of three spiking levels( 1. 0,10. 0 and 100. 0 μg /kg) ranged from 86. 9% to 102. 0%,and the relative standard deviations of 1. 80%- 4. 19% were obtained. Conclusion The method for determination of 3-nitropropionic acid in poisoning samples by UPLC-MS / MS is of operation convenience,less interference from impurities and good accuracy,which could provide evidence and treatment for mouldy sugarcane poisoning.更多还原