QuEChERS-超高效液相色谱-串联质谱法测定血液中4种乌头类生物碱
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摘要
移取1.00mL血液样品,加入2mL乙腈和50mg氯化钠,振荡10min后,在-4℃下以8 000r·min~(-1)转速离心10min,取上清液,加入15mg N-丙基乙二胺和25mg十八烷基硅烷,振荡5min,重复上述离心操作,取上清液,过0.22μm有机微孔膜,采用超高效液相色谱-串联质谱法测定滤液中次乌头碱、新乌头碱、乌头碱和滇乌头碱等4种乌头类生物碱的含量。以Agilent ZORBAX Eclipse Plus C_(18)色谱柱为固定相,以不同体积比的0.1%(质量分数)氨水和甲醇的混合液为流动相进行梯度洗脱,串联质谱分析中采用电喷雾正离子源和多反应监测模式。4种乌头类生物碱的质量浓度在一定范围内与其对应的峰面积呈线性关系,方法的检出限(3S/N)为0.010～0.035μg·L~(-1)。以空白样品为基体进行加标回收试验,所得回收率为96.3%～109%,回收量的日内相对标准偏差(n=6)为2.3%～4.2%,日间相对标准偏差(n=6)为0.70%～6.7%。
The blood sample(1.00 mL) was taken,and then 2 mL of acetonitrile and 50 mg of sodium chloride were added.After oscillation for 10 min,the above mixture was centrifugated for 10 min at the speed of 8 000 r·min~(-1) at -4℃.PSA(15 mg) and C_(18)(25 mg) were added in the supernatant.After oscillation for 5 min,the above centrifugal operation was repeated.The supernatant was filtered on 0.22 μm organic microporous membrane.UHPLC-MS/MS was applied to the determination of 4 aconitum alkaloids,i.e.hypaconitine,mesaconine,aconitine and yunaconitine in the filtrate.Agilent ZORBAX Eclipse Plus C_(18) chromatographic column was used as stationary phase,and the mixture of 0.1%(mass fraction) ammonia and methanol mixed in different ratios was used as mobile phase in gradient elution.ESI+and multi-reactions monitor mode were adopted in MS/MS.Linear relationships between values of peak area and mass concentration of the 4 aconitum alkaloids were kept in definite ranges.Detection limits(3S/N) of the method ranged from 0.010 to 0.035μg·L~(-1).On the base of blank sample,test for recovery was made by standard addition method;values of recovery found were in the range of 96.3%-109%,with intra-day RSD(n=6) and inter-day RSD(n=6) of recovery amounts in the ranges of 2.3%-4.2% and 0.70%-6.7%.
The blood sample(1.00 mL) was taken,and then 2 mL of acetonitrile and 50 mg of sodium chloride were added.After oscillation for 10 min,the above mixture was centrifugated for 10 min at the speed of 8 000 r·min~(-1) at -4℃.PSA(15 mg) and C_(18)(25 mg) were added in the supernatant.After oscillation for 5 min,the above centrifugal operation was repeated.The supernatant was filtered on 0.22 μm organic microporous membrane.UHPLC-MS/MS was applied to the determination of 4 aconitum alkaloids,i.e.hypaconitine,mesaconine,aconitine and yunaconitine in the filtrate.Agilent ZORBAX Eclipse Plus C_(18) chromatographic column was used as stationary phase,and the mixture of 0.1%(mass fraction) ammonia and methanol mixed in different ratios was used as mobile phase in gradient elution.ESI+and multi-reactions monitor mode were adopted in MS/MS.Linear relationships between values of peak area and mass concentration of the 4 aconitum alkaloids were kept in definite ranges.Detection limits(3S/N) of the method ranged from 0.010 to 0.035μg·L~(-1).On the base of blank sample,test for recovery was made by standard addition method;values of recovery found were in the range of 96.3%-109%,with intra-day RSD(n=6) and inter-day RSD(n=6) of recovery amounts in the ranges of 2.3%-4.2% and 0.70%-6.7%.
引文
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[4]杨发俊.45例急性乌头碱中毒的临床表现和救治分析[J].中国医学创新,2017,14(36):94-97.
[5]粟贵,廖林川,颜有仪,等.HPLC同时测定生物样品中新乌头碱、乌头碱、次乌头碱的含量[J].中国药学杂志,2009,44(12):946-950.
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[11] BEIKE J,FROMMHERZ L,WOOD M,et al.Determination of aconitine in body fluids by LC-MS-MS[J].International Journal of Legal Medicine,2004,118(5):289-293.
[12] KA-WING CHUNG K,PAK-LAM CHEN S,NG S W,et al.Measurement of yunaconitine and crassicauline A in small-volume blood serum samples by LCMS/MS:Tracing of aconite poisoning in clinical diagnosis[J].Talanta,2012,97:491-498.
[13]朱定姬,卢敏萍,黄克建,等.在线固相萃取结合液相色谱-线性离子阱多级质谱法同时检测生物样品中7种乌头类生物碱[J].色谱,2016,34(3):249-257.
[14]武洁,沈红,朱玲英,等.液-质联用法同时测定大鼠血浆中的乌头碱、新乌头碱、次乌头碱及其药动学[J].中国医院药学杂志,2011,31(14):1162-1166.
[15]张润生,余琛,刘罡一,等.血液中乌头碱、次乌头碱、新乌头碱的LC/MS/MS分析[J].中国法医学杂志,2004,19(5):265-267.
[16] 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.
[17]姚芳,宋志宇,聂晶,等.气相色谱-质谱法结合方法和分散液液微萃取技术测定茭白中三唑类农药残留[J].理化检验-化学分册,2017,53(6):650-654.
[18]陈刚,邓晓军,孙锦兰,等.高效液相色谱-串联质谱法同时测定动物源性食品中35种兽药残留量[J].理化检验-化学分册,2014,50(7):809-814.
[19]熊欣,刘青,张广文,等.QuEChERS/LC-MS/MS法测定食品中7种真菌毒素[J].分析测试学报,2018,37(9):1008-1013.
[20]董芳,李惠玲,马婧,等.全血中有机磷农药和鼠药测定的QuEChERS前处理法[J].中华劳动卫生职业病杂志,2016,34(10):782-785.
[21] DULAURENT S,EL BALKHI S,PONCELET L,et al.QuEChERS sample preparation prior to LCMS/MS determination of opiates,amphetamines,and cocaine metabolites in whole blood[J].Analytical and Bioanalytical Chemistry,2016,408(5):1467-1474.
[22]徐多麒,张蕾萍,王继芬,等.超高效液相色谱串联质谱法同时快速检验全血中扎来普隆及其代谢产物[J].分析化学,2016,44(7):1059-1064.
[23]于天晓,韩彦洁,张瑶,等.基于Fe3O4磁性纳米粒子的QuEChERS预处理方法联合GC-MS对血液中多种苯二氮卓类药物检测[J].分析试验室,2018,37(4):450-453.
[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.
[25]张洪非,罗彦波,李翔宇,等.改进QuEChERS-气相色谱/质谱法测定食源性植物中烟碱及次要生物碱[J].分析科学学报,2018,34(2):239-244.
[26]张毅,翁代群,王海军,等.乌头碱的水解动力学研究[J].中草药,2009,40(增刊1):120-122.
[27]刘婷.QuEChERS结合GC-MS/MS和LC-MS/MS高通量检测果蔬农药多残留研究[D].长沙:中南林业科技大学,2018.
[2]刘帅,李妍,李卫飞,等.乌头类中药毒性及现代毒理学研究进展[J].中草药,2016,47(22):4095-4102.
[3]杨苗苗,杨霖,李孝栋.乌头安全有效性研究进展及其临床应用前景[J].药学研究,2018,37(3):125-133.
[4]杨发俊.45例急性乌头碱中毒的临床表现和救治分析[J].中国医学创新,2017,14(36):94-97.
[5]粟贵,廖林川,颜有仪,等.HPLC同时测定生物样品中新乌头碱、乌头碱、次乌头碱的含量[J].中国药学杂志,2009,44(12):946-950.
[6]黄勤安,张聿梅,何轶,等.乌头碱水解转化规律的研究[J].中国中药杂志,2007,32(20):2143-2145.
[7]王朝虹,何毅,张继宗,等.生物检材中乌头碱的GC/MS分析[J].中国法医学杂志,2003,18(3):145-146.
[8] WANG Z H,GUO D A,HE Y,et al.Quantitative determination of aconitum alkaloids in blood and urine samples by high-performance liquid chromatography[J].Phytochemical Analysis,2004,15(1):16-20.
[9]滕傲雪,王瑞花,何洪源,等.超高效液相色谱-串联质谱法同时测定血液中5种乌头类生物碱[J].理化检验-化学分册,2018,54(8):941-945.
[10] ZHANG F,TANG M H,CHEN L J,et al.Simultaneous quantitation of aconitine,mesaconitine,hypaconitine, benzoylaconine, benzoylmesaconine and benzoylhypaconine in human plasma by liquid chromatography-tandem mass spectrometry and pharmacokinetics evaluation of“SHEN-FU”injectable powder[J].Journal of Chromatography B,2008,873(2):173-179.
[11] BEIKE J,FROMMHERZ L,WOOD M,et al.Determination of aconitine in body fluids by LC-MS-MS[J].International Journal of Legal Medicine,2004,118(5):289-293.
[12] KA-WING CHUNG K,PAK-LAM CHEN S,NG S W,et al.Measurement of yunaconitine and crassicauline A in small-volume blood serum samples by LCMS/MS:Tracing of aconite poisoning in clinical diagnosis[J].Talanta,2012,97:491-498.
[13]朱定姬,卢敏萍,黄克建,等.在线固相萃取结合液相色谱-线性离子阱多级质谱法同时检测生物样品中7种乌头类生物碱[J].色谱,2016,34(3):249-257.
[14]武洁,沈红,朱玲英,等.液-质联用法同时测定大鼠血浆中的乌头碱、新乌头碱、次乌头碱及其药动学[J].中国医院药学杂志,2011,31(14):1162-1166.
[15]张润生,余琛,刘罡一,等.血液中乌头碱、次乌头碱、新乌头碱的LC/MS/MS分析[J].中国法医学杂志,2004,19(5):265-267.
[16] 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.
[17]姚芳,宋志宇,聂晶,等.气相色谱-质谱法结合方法和分散液液微萃取技术测定茭白中三唑类农药残留[J].理化检验-化学分册,2017,53(6):650-654.
[18]陈刚,邓晓军,孙锦兰,等.高效液相色谱-串联质谱法同时测定动物源性食品中35种兽药残留量[J].理化检验-化学分册,2014,50(7):809-814.
[19]熊欣,刘青,张广文,等.QuEChERS/LC-MS/MS法测定食品中7种真菌毒素[J].分析测试学报,2018,37(9):1008-1013.
[20]董芳,李惠玲,马婧,等.全血中有机磷农药和鼠药测定的QuEChERS前处理法[J].中华劳动卫生职业病杂志,2016,34(10):782-785.
[21] DULAURENT S,EL BALKHI S,PONCELET L,et al.QuEChERS sample preparation prior to LCMS/MS determination of opiates,amphetamines,and cocaine metabolites in whole blood[J].Analytical and Bioanalytical Chemistry,2016,408(5):1467-1474.
[22]徐多麒,张蕾萍,王继芬,等.超高效液相色谱串联质谱法同时快速检验全血中扎来普隆及其代谢产物[J].分析化学,2016,44(7):1059-1064.
[23]于天晓,韩彦洁,张瑶,等.基于Fe3O4磁性纳米粒子的QuEChERS预处理方法联合GC-MS对血液中多种苯二氮卓类药物检测[J].分析试验室,2018,37(4):450-453.
[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.
[25]张洪非,罗彦波,李翔宇,等.改进QuEChERS-气相色谱/质谱法测定食源性植物中烟碱及次要生物碱[J].分析科学学报,2018,34(2):239-244.
[26]张毅,翁代群,王海军,等.乌头碱的水解动力学研究[J].中草药,2009,40(增刊1):120-122.
[27]刘婷.QuEChERS结合GC-MS/MS和LC-MS/MS高通量检测果蔬农药多残留研究[D].长沙:中南林业科技大学,2018.