超高效液相色谱-串联质谱法检验腐败血中吗啡和6-单乙酰吗啡
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摘要
采用改良的QuEChERS前处理方法,建立了同时检验腐败血中吗啡和6-单乙酰吗啡的超高效液相色谱-串联质谱(UPLC-MS/MS)方法。腐败血中加入乙腈-水(4∶1,v/v)混合溶液、30 mg NaCl和60 mg MgSO4盐析促进相分离,最后经25 mg N-丙基乙二胺(PSA)和25 mg MgSO4净化。选用ZORBAX Eclipse Plus C18色谱柱分离,0. 01%(v/v)氨水和乙腈作为流动相进行梯度洗脱,电喷雾电离正离子(ESI+)模式扫描,多反应监测(MRM)模式检测。结果表明,吗啡和6-单乙酰吗啡在5～200μg/L范围内线性关系良好,相关系数(r2)≥0. 995 7,检出限(S/N=3)均为1μg/L,定量限(S/N=10)均为5μg/L。3个加标水平(5、100和200μg/L)下,吗啡和6-单乙酰吗啡的平均加标回收率分别为81. 84%～103. 44%和81. 03%～104. 46%,日内和日间精密度(RSD)均小于12%,基质效应为83. 04%～107. 61%。方法简便、灵敏、可靠,可实现腐败血中吗啡和6-单乙酰吗啡的快速定性定量检验。
This study was performed to develop an ultra-performance liquid chromatography tandem mass spectrometry( UPLC-MS/MS) method to detect and quantify morphine and 6-monoacetylmorphine in putrefied blood for forensic toxicological purposes. A modified QuEChERS method was implemented as follows: extraction of morphine and 6-monoacetylmorphine from putrefied blood was performed with an acetonitrile-water( 4 ∶1,v/v) mixture,and 30 mg NaCl and 60 mg anhydrous MgSO4 were subsequently added as salting out agents to induce phase separation.The supernatant was cleaned by adding 25 mg primary secondary amine sorbent( PSA)and 25 mg anhydrous MgSO4. Separation of the target compounds was performed using a ZORBAX Eclipse Plus C18 column by UPLC over a 4 min gradient elution where 0. 01%( v/v) ammonium hydroxide buffer( A) and acetonitrile( B) were used as the mobile phase. MS/MS was used in positive electrospray ionization( ESI+) mode and multiple-reaction monitoring( MRM) was performed to detect the target drugs. This method achieved satisfactory recoveries( R) for morphine and 6-monoacetylmorphine with mean recovery values ranging from 81. 84% to 103. 44%and 81. 03% to 104. 46% respectively. The developed method also provided efficient purification of the sample from endogenous interferences with matrix effect( ME) ranging from 83. 04% to107. 61%. The method was validated and the limit of detection( LOD) and limit of quantification( LOQ) were 1 and 5 μg/L,respectively,for both compounds and the precision with RSD ranged from 1% to 12%. This method proved to be quick,sensitive,rugged,and suitable for the analysis of illegal drugs in putrefied whole blood.
This study was performed to develop an ultra-performance liquid chromatography tandem mass spectrometry( UPLC-MS/MS) method to detect and quantify morphine and 6-monoacetylmorphine in putrefied blood for forensic toxicological purposes. A modified QuEChERS method was implemented as follows: extraction of morphine and 6-monoacetylmorphine from putrefied blood was performed with an acetonitrile-water( 4 ∶1,v/v) mixture,and 30 mg NaCl and 60 mg anhydrous MgSO4 were subsequently added as salting out agents to induce phase separation.The supernatant was cleaned by adding 25 mg primary secondary amine sorbent( PSA)and 25 mg anhydrous MgSO4. Separation of the target compounds was performed using a ZORBAX Eclipse Plus C18 column by UPLC over a 4 min gradient elution where 0. 01%( v/v) ammonium hydroxide buffer( A) and acetonitrile( B) were used as the mobile phase. MS/MS was used in positive electrospray ionization( ESI+) mode and multiple-reaction monitoring( MRM) was performed to detect the target drugs. This method achieved satisfactory recoveries( R) for morphine and 6-monoacetylmorphine with mean recovery values ranging from 81. 84% to 103. 44%and 81. 03% to 104. 46% respectively. The developed method also provided efficient purification of the sample from endogenous interferences with matrix effect( ME) ranging from 83. 04% to107. 61%. The method was validated and the limit of detection( LOD) and limit of quantification( LOQ) were 1 and 5 μg/L,respectively,for both compounds and the precision with RSD ranged from 1% to 12%. This method proved to be quick,sensitive,rugged,and suitable for the analysis of illegal drugs in putrefied whole blood.
引文
[1] Nations Office on Drugs and Crime. World Drug Report 2017.(2017-06-22)[2018-07-11]. http://w w w. unodc. org/w dr2017/field/Bo o klet_2_HEALTH.pdf
[2] Nations Office on Drugs and Crime. World Drug Report 2018.(2018-06-26)[2018-07-11]. https://w w w. unodc. org/w dr2018/prelaunch/WDR18_Bo o klet_2_GLOBAL.pdf
[3] Saw ynok J. Can J Physiol Pharmacol,1986,64(1):1
[4] Klous M G,Brink W V,Van Ree J M,et al. Drug Alcohol Depen,2005,80(3):283
[5] Anastassiades M,Lehotay S J,Stajnbaher D,et,al. J AOAC Int,2003,86(2):412
[6] Bian Z Y,Deng H M. QuEChERS Technology and Applicatio n. Beijing:China Light Industry Press,2017边照阳,邓慧敏. QuEChERS技术及应用.北京:中国轻工业出版社,2017
[7] Lehotay S J,Mastovska K,Yun S J. J AOAC Int,2005,88(2):630
[8] Wang L Z,Zhou Y,Huang X Y,et al. Chinese Journal of Chro m ato graphy,2013,31(12):1167王连珠,周昱,黄小燕,等.色谱,2013,31(12):1167
[9] Zheng L,Wu Y J,Zhao Y F,et al. Chinese Journal of Chrom ato graphy,2014,32(8):867郑玲,吴玉杰,赵永锋,等.色谱,2014,32(8):867
[10] Miao Q C,Nie J,Wu H Z,et al. Physical Testing and Chem ical Analysis:Part B(Chem ical Analysis),2017,53(7):745苗琼晨,聂晶,吴惠珍,等.理化检验-化学分册,2017,53(7):745
[11] Liu Y Q,Liu S,Xu W J,et al. Chinese Journal of Chromato graphy,2017,35(9):941刘永强,刘胜,许文娟,等.色谱,2017,35(9):941
[12] Zheng J,Xi C X,Cao S R,et al. Chinese Journal of Chrom ato graphy,2017,35(12):1257郑佳,郗存显,曹淑瑞,等.色谱,2017,35(12):1257
[13] Wang S W,Liu Y P,Sun H B,et al. Chinese Journal of Chro m ato graphy,2018,36(1):17王思威,刘艳萍,孙海滨,等.色谱,2018,36(1):17
[14] Plo¨ssl F,Giera M,Bracher F. J Chromatogr A,2006,1135(1):19
[15] Guo Z,Chang J,Yang R Q,et al. Journal of Instrumental Analysis,2015,34(9):986郭震,常靖,杨瑞琴,等.分析测试学报,2015,34(9):986
[16] Yu T,Wang T,Huang Z,et al. Chromatographia,2016,80(1):1
[17] Xu D Q,Zhang L P,Wang J F,et al. Chinese Journal of Analysis Chem istry,2016,44(7):1059徐多麒,张蕾萍,王继芬,等.分析化学,2016,44(7):1059
[18] Lin K,Wang J F,Lv Y F,et al. Chinese Journal of Analysis Labo rato ry,2017,36(8):894林宽,王继芬,吕昱帆,等.分析试验室,2017,36(8):894
[19] Allen D L,Oliver J S. J. Anal Toxicol,2000,24(3):228
[20] Stimpfl T,Jurenitsch J,Vycudilik W. J Anal Toxicol,2001,25(2):125
[21] Pouliopoulos A,Tsakelidou E,Krokos A,et al. J Anal To x ico l,2018,42(5):337
[22] Qiu Y M,Zhu J,Chang J,et al. Forensic Science and Techno lo gy,2010,35(1):3邱玉敏,朱军,常靖,等.刑事技术,2010,35(1):3
[23] Lehmann S,Schulze B,Thomas A,et al. Forensic Toxico l,2018,36(2):320
[24] Lv Y F,Wang J F,Li C,et al. Chinese Journal of Analysis Labo rato ry,2018,37(9):998吕昱帆,王继芬,李超,等.分析试验室,2018,37(9):998
[25] Matuszew ski B,Constanzer M,Chavez-Eng C. Anal Chem,2003,75(13):3019
[26] Rook E J,Huitema A D R,Brink W V,et al. Clin Pharmaco kinet,2006,45(4):401
[27] Chen Y,Zhu J,Zhang Y F,et al. Chinese Journal of Forensic Medicine,2016,31(1):9陈跃,朱军,张云峰,等.中国法医学杂志,2016,31(1):9
[28] Yu X W,Wang M,Tan X H,et al. Chinese Journal of Forensic Medicine,2014,29(3):235于晓巍,王玫,谭晓辉,等.中国法医学杂志,2014,29(3):235
[2] Nations Office on Drugs and Crime. World Drug Report 2018.(2018-06-26)[2018-07-11]. https://w w w. unodc. org/w dr2018/prelaunch/WDR18_Bo o klet_2_GLOBAL.pdf
[3] Saw ynok J. Can J Physiol Pharmacol,1986,64(1):1
[4] Klous M G,Brink W V,Van Ree J M,et al. Drug Alcohol Depen,2005,80(3):283
[5] Anastassiades M,Lehotay S J,Stajnbaher D,et,al. J AOAC Int,2003,86(2):412
[6] Bian Z Y,Deng H M. QuEChERS Technology and Applicatio n. Beijing:China Light Industry Press,2017边照阳,邓慧敏. QuEChERS技术及应用.北京:中国轻工业出版社,2017
[7] Lehotay S J,Mastovska K,Yun S J. J AOAC Int,2005,88(2):630
[8] Wang L Z,Zhou Y,Huang X Y,et al. Chinese Journal of Chro m ato graphy,2013,31(12):1167王连珠,周昱,黄小燕,等.色谱,2013,31(12):1167
[9] Zheng L,Wu Y J,Zhao Y F,et al. Chinese Journal of Chrom ato graphy,2014,32(8):867郑玲,吴玉杰,赵永锋,等.色谱,2014,32(8):867
[10] Miao Q C,Nie J,Wu H Z,et al. Physical Testing and Chem ical Analysis:Part B(Chem ical Analysis),2017,53(7):745苗琼晨,聂晶,吴惠珍,等.理化检验-化学分册,2017,53(7):745
[11] Liu Y Q,Liu S,Xu W J,et al. Chinese Journal of Chromato graphy,2017,35(9):941刘永强,刘胜,许文娟,等.色谱,2017,35(9):941
[12] Zheng J,Xi C X,Cao S R,et al. Chinese Journal of Chrom ato graphy,2017,35(12):1257郑佳,郗存显,曹淑瑞,等.色谱,2017,35(12):1257
[13] Wang S W,Liu Y P,Sun H B,et al. Chinese Journal of Chro m ato graphy,2018,36(1):17王思威,刘艳萍,孙海滨,等.色谱,2018,36(1):17
[14] Plo¨ssl F,Giera M,Bracher F. J Chromatogr A,2006,1135(1):19
[15] Guo Z,Chang J,Yang R Q,et al. Journal of Instrumental Analysis,2015,34(9):986郭震,常靖,杨瑞琴,等.分析测试学报,2015,34(9):986
[16] Yu T,Wang T,Huang Z,et al. Chromatographia,2016,80(1):1
[17] Xu D Q,Zhang L P,Wang J F,et al. Chinese Journal of Analysis Chem istry,2016,44(7):1059徐多麒,张蕾萍,王继芬,等.分析化学,2016,44(7):1059
[18] Lin K,Wang J F,Lv Y F,et al. Chinese Journal of Analysis Labo rato ry,2017,36(8):894林宽,王继芬,吕昱帆,等.分析试验室,2017,36(8):894
[19] Allen D L,Oliver J S. J. Anal Toxicol,2000,24(3):228
[20] Stimpfl T,Jurenitsch J,Vycudilik W. J Anal Toxicol,2001,25(2):125
[21] Pouliopoulos A,Tsakelidou E,Krokos A,et al. J Anal To x ico l,2018,42(5):337
[22] Qiu Y M,Zhu J,Chang J,et al. Forensic Science and Techno lo gy,2010,35(1):3邱玉敏,朱军,常靖,等.刑事技术,2010,35(1):3
[23] Lehmann S,Schulze B,Thomas A,et al. Forensic Toxico l,2018,36(2):320
[24] Lv Y F,Wang J F,Li C,et al. Chinese Journal of Analysis Labo rato ry,2018,37(9):998吕昱帆,王继芬,李超,等.分析试验室,2018,37(9):998
[25] Matuszew ski B,Constanzer M,Chavez-Eng C. Anal Chem,2003,75(13):3019
[26] Rook E J,Huitema A D R,Brink W V,et al. Clin Pharmaco kinet,2006,45(4):401
[27] Chen Y,Zhu J,Zhang Y F,et al. Chinese Journal of Forensic Medicine,2016,31(1):9陈跃,朱军,张云峰,等.中国法医学杂志,2016,31(1):9
[28] Yu X W,Wang M,Tan X H,et al. Chinese Journal of Forensic Medicine,2014,29(3):235于晓巍,王玫,谭晓辉,等.中国法医学杂志,2014,29(3):235