QuEChERS提取-超高效液相色谱-串联质谱法测定腐败人血中3种钩吻生物碱
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摘要
为同时测定腐败血液中钩吻生物碱(钩吻素子、钩吻素甲和钩吻素己等3种),应用QuEChERS前处理方法提取上述3种分析物,并用超高效液相色谱-串联质谱法进行测定。取血样1.00mL,加入乙腈2mL和NaCl 50mg,涡旋振荡提取20s,继续振荡10min后,以8 000r·min~(-1)转速离心10min。取上清液,加入十八烷基硅烷(C_(18))20mg作为吸附剂,按上述相同方法振荡并离心,进行基质分散固相提取。取上清液按仪器工作条件进行分析。在色谱分离中,应用ZORBAX Eclipse Plus C18色谱柱为固定相和不同比例的(A)乙腈和(B)含φ0.1%甲酸的5mmol·L~(-1)甲酸铵溶液的混合溶液为流动相进行梯度淋洗,达到上述3种钩吻生物碱的良好分离。质谱测定中选择电喷雾正离子源和多反应监测模式。3种钩吻生物碱的质量浓度在一定范围内与对应的峰面积呈线性关系,检出限(3S/N)在0.005～0.1μg·L~(-1)之间。在空白血样的基础上加入上述3种钩吻生物碱的混合标准溶液,并按上述方法测定求得其回收率在90.5%～110%之间,测定值的相对标准偏差(n=6)在2.1%～5.1%。
Three of the gelsemium alkaloids in putrefied whole blood were pretreated by the QuEChERS approach and determined by UPLC-MS/MS.In the QuEChERS,1.00 mL of the blood sample was extracted by swirly shaking for 20 swith 2 mL of acetonitrile and 50 mg of NaCl,and the shaking was continued for 10 min,and the mixture was centrifuged for 10 min at a rate of 8 000 r·min~(-1).The supernatant was taken and treated by the dispersed-matrix SPE using 20 mg of C_(18) as adsorbent and the mixture was shaken and centrifuged in the same way as described above.The supernatant was separated and used for UPLC and MS/MS analysis as described in the instrumental working conditions.The chromatographic column of ZORBAX Eclipse Plus C_(18) was selected as stationary phase,and mixtures of(A)acetonitrile and(B)5 mmol·L~(-1) ammonium formate(containingφ0.1%of formic acid)in various ratios were used as mobile phase in gradient elution,to have the 3 gelsemium alkaloids wellseparated.In the MS/MS determinations,electrospray positive ion source and mode of multi-reaction monitoring were adopted in MS/MS.Linear relationships were obtained between values of peak areas and mass concentrations of the 3 gelsemium alkaloids in definite ranges,with detection limits(3 S/N)in the range of 0.005-0.1μg·L~(-1).Recovery tests were made by addition mixed standard solutions of the 3 gelsemium alkaloids to a blank sample as matrix and then determined by the proposed method,giving values of recovery in the range of 90.5%~(-1)10%,and values of RSDs(n=6)in the range of 2.1%-5.1%.
Three of the gelsemium alkaloids in putrefied whole blood were pretreated by the QuEChERS approach and determined by UPLC-MS/MS.In the QuEChERS,1.00 mL of the blood sample was extracted by swirly shaking for 20 swith 2 mL of acetonitrile and 50 mg of NaCl,and the shaking was continued for 10 min,and the mixture was centrifuged for 10 min at a rate of 8 000 r·min~(-1).The supernatant was taken and treated by the dispersed-matrix SPE using 20 mg of C_(18) as adsorbent and the mixture was shaken and centrifuged in the same way as described above.The supernatant was separated and used for UPLC and MS/MS analysis as described in the instrumental working conditions.The chromatographic column of ZORBAX Eclipse Plus C_(18) was selected as stationary phase,and mixtures of(A)acetonitrile and(B)5 mmol·L~(-1) ammonium formate(containingφ0.1%of formic acid)in various ratios were used as mobile phase in gradient elution,to have the 3 gelsemium alkaloids wellseparated.In the MS/MS determinations,electrospray positive ion source and mode of multi-reaction monitoring were adopted in MS/MS.Linear relationships were obtained between values of peak areas and mass concentrations of the 3 gelsemium alkaloids in definite ranges,with detection limits(3 S/N)in the range of 0.005-0.1μg·L~(-1).Recovery tests were made by addition mixed standard solutions of the 3 gelsemium alkaloids to a blank sample as matrix and then determined by the proposed method,giving values of recovery in the range of 90.5%~(-1)10%,and values of RSDs(n=6)in the range of 2.1%-5.1%.
引文
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[9]吴惠勤，张春华，黄晓兰，等．气相色谱-串联质谱法同时检测尿液中15种有毒生物碱[J]．分析测试学报，2013,32(9):1031-1037.
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[12]熊小婷，吴惠勤，黄晓兰．液相色谱-电喷雾串联质谱同时检测血液中8种有毒生物碱[J]．分析化学，2009,37(10):1433-1438.
[13]宋蕊，武继锋，刘海燕，等．HPLC-MS/MS法快速检测人血中钩吻毒素[J]．中国刑警学院学报，2017(2):103-106.
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[15] SCHENCK F J, HOBBS J E.Evaluation of the quick,easy,cheap,effective,rugged,and safe(QuEChERS)approach to pesticide residue analysis[J].Bulletin of Environmental Contamination and Toxicology,2004,73(1):24-30.
[16] GONZLEZ-JARTN J M,ALFONSO A,RODRGUEZ I,et al.A QuEChERS based extraction procedure coupled to UPLC-MS/MS detection for mycotoxins analysis in beer[J].Food Chemistry,2019,275:703-710.
[17] DUAN X Y,TONG L,LI D X,et al.A multiresidue method for simultaneous determination of 116pesticides in notoginseng radix et rhizoma using modified QuEChERS coupled with gas chromatography tandem mass spectrometry and census 180batches of sample from yunnan province[J].Chromatographia,2018,81(3):545-556.
[18] XU X,XU X Y,HAN M,et al.Development of a modified QuEChERS method based on magnetic multiwalled carbon nanotubes for the simultaneous determination of veterinary drugs,pesticides and mycotoxins in eggs by UPLC-MS/MS[J].Food Chemistry,2019,276:419-426.
[19] JUAN C,MAES J,FONT G,et al.Determination of mycotoxins in fruit berry by-products using QuEChERS extraction method[J].LWT-Food Science and Technology,2017,86:344-351.
[20]万超超，任昕昕，程芳彬，等．QuEChERS-超高效液相色谱-串联质谱法同时测定人全血中5种氨基甲酸酯类农药[J]．理化检验-化学分册，2018,54(12):1383-1388.
[21]黄伟华，夏祥，胡美华．QuEChERS-在线GPC-GC/MS联用法测定尿液中11种苯二氮卓类药物[J]．分析试验室，2019,38(1):48-51.
[22]吕昱帆，王继芬，李超，等．QuEChERS结合超高效液相色谱-串联质谱法检验腐败血中4-甲基甲卡西酮和甲卡西酮[J]．分析试验室，2018,37(9):998-1002.
[23]宋强．硬聚氯乙烯管材耐丙酮测定方法[J]．塑料科技，1989,17(1):37-38.
[24] REJCZAK T,TUZIMSKI T.A review of recent developments and trends in the QuEChERS sample preparation approach[J].Open Chemistry,2015,13(1):980-1010.
[2]杜秀宝，戴韵华，张常麟，等．钩吻生物碱的研究-Ⅰ．钩吻素己的结构[J]．化学学报，1982,40(12):1137-1141.
[3]孟东江，罗友杰．抢救8例钩吻中毒的经验及教训[J]．中国社区医师（医学专业），2012,14(16):315-315.
[4]杨靓．急性药物中毒的临床表现[J]．中国社区医师，2007,23(17):3-4.
[5]马进，胡丙杰．大鼠死后细胞DNA的降解变化与死亡时间的关系[J]．广东医学，2018,39(14):2110-2113.
[6] DONALDSON A E,LAMONT I L.Biochemistry changes that occur after death:Potential markers for determining post-mortem interval[J].Plos One,2013,8(11):1-10.
[7]刘兆，杨瑞琴，温永启，等．腐败血液中乙醇的顶空气相色谱分析[J]．中国司法鉴定，2010(4):26-28.
[8]马克芹，王毅．GC/MS法测定人血中钩吻碱甲[J]．微量元素与健康研究，2014,31(6):48-50.
[9]吴惠勤，张春华，黄晓兰，等．气相色谱-串联质谱法同时检测尿液中15种有毒生物碱[J]．分析测试学报，2013,32(9):1031-1037.
[10]丘宏强，程昱，刘茂柏，等．人血浆中钩吻素甲和钩吻素子HPLC-UV测定[J]．中草药，2016,47(13):2324-2327.
[11]姬圣洁，刘伟．LC-MS/MS法同时检测生物检材中钩吻素子、钩吻素甲和钩吻素己[J]．法医学杂志，2017,33(2):141-147.
[12]熊小婷，吴惠勤，黄晓兰．液相色谱-电喷雾串联质谱同时检测血液中8种有毒生物碱[J]．分析化学，2009,37(10):1433-1438.
[13]宋蕊，武继锋，刘海燕，等．HPLC-MS/MS法快速检测人血中钩吻毒素[J]．中国刑警学院学报，2017(2):103-106.
[14] CHEN J Z,LI Y,XIAO J P,et al.Development of a sensitive and rapid UPLC-MS/MS method for the determination of koumine in rat plasma:Application to apharmacokinetic study[J].Biomedical Chromatography,2013,27(6):736-740.
[15] SCHENCK F J, HOBBS J E.Evaluation of the quick,easy,cheap,effective,rugged,and safe(QuEChERS)approach to pesticide residue analysis[J].Bulletin of Environmental Contamination and Toxicology,2004,73(1):24-30.
[16] GONZLEZ-JARTN J M,ALFONSO A,RODRGUEZ I,et al.A QuEChERS based extraction procedure coupled to UPLC-MS/MS detection for mycotoxins analysis in beer[J].Food Chemistry,2019,275:703-710.
[17] DUAN X Y,TONG L,LI D X,et al.A multiresidue method for simultaneous determination of 116pesticides in notoginseng radix et rhizoma using modified QuEChERS coupled with gas chromatography tandem mass spectrometry and census 180batches of sample from yunnan province[J].Chromatographia,2018,81(3):545-556.
[18] XU X,XU X Y,HAN M,et al.Development of a modified QuEChERS method based on magnetic multiwalled carbon nanotubes for the simultaneous determination of veterinary drugs,pesticides and mycotoxins in eggs by UPLC-MS/MS[J].Food Chemistry,2019,276:419-426.
[19] JUAN C,MAES J,FONT G,et al.Determination of mycotoxins in fruit berry by-products using QuEChERS extraction method[J].LWT-Food Science and Technology,2017,86:344-351.
[20]万超超，任昕昕，程芳彬，等．QuEChERS-超高效液相色谱-串联质谱法同时测定人全血中5种氨基甲酸酯类农药[J]．理化检验-化学分册，2018,54(12):1383-1388.
[21]黄伟华，夏祥，胡美华．QuEChERS-在线GPC-GC/MS联用法测定尿液中11种苯二氮卓类药物[J]．分析试验室，2019,38(1):48-51.
[22]吕昱帆，王继芬，李超，等．QuEChERS结合超高效液相色谱-串联质谱法检验腐败血中4-甲基甲卡西酮和甲卡西酮[J]．分析试验室，2018,37(9):998-1002.
[23]宋强．硬聚氯乙烯管材耐丙酮测定方法[J]．塑料科技，1989,17(1):37-38.
[24] REJCZAK T,TUZIMSKI T.A review of recent developments and trends in the QuEChERS sample preparation approach[J].Open Chemistry,2015,13(1):980-1010.
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