Article(id=1239250703600906490, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239250701583446236, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024.04.07, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1687449600000, receivedDateStr=2023-06-23, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773389991902, onlineDateStr=2026-03-13, pubDate=1714406400000, pubDateStr=2024-04-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773389991902, onlineIssueDateStr=2026-03-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773389991902, creator=13701087609, updateTime=1773389991902, updator=13701087609, issue=Issue{id=1239250701583446236, tenantId=1146029695717560320, journalId=1205117023404326918, year='2024', volume='44', issue='4', pageStart='553', pageEnd='736', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773389991422, creator=13701087609, updateTime=1773390513217, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1239252890213209050, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239250701583446236, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1239252890213209051, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239250701583446236, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=603, endPage=609, ext={EN=ArticleExt(id=1239250705148604680, articleId=1239250703600906490, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Detection of dextromethorphan and dextrorphan in hair by UPLC-MS/MS*, columnId=1239184757519602223, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Metabolism Analysis, runingTitle=null, highlight=null, articleAbstract=
Objective:

To establish a rapid and accurate ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the detection of dextromethorphan and one metabolite dextrorphan in hair.

Methods:

The hair sample containing dextromethorphan and dextrorphan were extracted with methanol containing internal standard proadifen hydrochloride(SKF525A). The extract was filtered with 0.22 μm organic filter membrane and detected by UPLC-MS/MS. All components were separated by an ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm), using a gradient elution procedure consisting of 0.2% formic acid (10 mmol·L-1 ammonium formate) and acetonitrile, at a flow rate of 0.3 mL·min-1, and the column temperature was room temperature. Positive electrospray ionization was performed using multiple reaction monitoring mode (MRM).

Results:

The linear relationships of dextromethorphan and dextrorphan were good in the range of 1-100 ng·mL-1. The linear equations were Y=1.349 49X-0.020 80 (r=0.998 8) and Y=0.775 10X-0.013 87 (r=0.999 1), respectively. The detection limit and quantitation limit were 0.010 ng·mL-1 and 0.025 ng·mL-1 respectively. Their recoveries ranged from 97.0%-104.8%. The intra-day RSD and inter-day RSD were 1.5%-3.9% and 2.1%-5.5%, respectively. The method was applied to cases, and the results showed that dextromethorphan and dextrorphan were detected in the hair of 6 abusers.

Conclusion:

This method is simple and sensitive enough to be applied to detect dextromethorphan and one metabolite dextrorphan in hair.

, correspAuthors=Li-chang GUAN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Nan ZHAO, Guo-liang ZHOU, Shu-he LI, Xu-dong ZHANG, Yuan TIAN, Li-chang GUAN), CN=ArticleExt(id=1239250705987465546, articleId=1239250703600906490, tenantId=1146029695717560320, journalId=1205117023404326918, language=CN, title=UPLC-MS/MS法检测人体毛发中右美沙芬及其代谢物去甲右美沙芬*, columnId=1239184757708345914, journalTitle=药物分析杂志, columnName=代谢分析, runingTitle=null, highlight=null, articleAbstract=
目的:

建立一种准确、快速检测毛发中右美沙芬及其代谢物去甲右美沙芬的超高效液相色谱-串联质谱(UPLC-MS/MS)方法。

方法:

用含内标双苯戊二氨酯(SKF525A)的甲醇溶液提取含有右美沙芬及其代谢物的毛发,经0.22 μm微孔有机滤膜过滤,UPLC-MS/MS检测。采用ACQUITY UPLC HSS T3超高液相色谱柱(100 mm×2.1 mm,1.8 μm),以0.2%甲酸(10 mmol·L-1甲酸铵)和乙腈为流动相进行梯度洗脱,流速0.3 mL·min-1,柱温为室温。采用电喷雾离子源,正离子多反应模式检测。

结果:

右美沙芬和去甲右美沙芬质量浓度均在1~100 ng·mL-1范围内线性响应良好,线性方程分别为Y=1.349 49X-0.020 80 (r=0.998 8)和Y=0.775 10X-0.013 87 (r=0.999 1),检测限和定量限均分别为0.01 ng·mL-1和0.025 ng·mL-1,平均回收率在97.0%~104.8%,日内精密度和日间精密度分别在1.5%~3.9%和2.1%~5.5%。将本方法应用于实际案例,结果在6例右美沙芬滥用者毛发中均检测出右美沙芬及其代谢物去甲右美沙芬。

结论:

该方法操作简便,检测灵敏度高,可用于毛发中右美沙芬及其代谢物去甲右美沙芬的检测。

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** Tel:18274898856;E-mail:
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articleTitle=null, refAbstract=null), Reference(id=1239250720621392827, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, doi=null, pmid=null, pmcid=null, year=2018, volume=null, issue=null, pageStart=5, pageEnd=null, url=null, language=null, rfNumber=[27], rfOrder=47, authorNames=null, journalName=null, refType=null, unstructuredReference=SF/Z JD0107025-2018 毛发中15种毒品及代谢物的液相色谱-串联质谱检验方法[S]. 2018:5, articleTitle=null, refAbstract=null), Reference(id=1239250720763999167, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, doi=null, pmid=null, pmcid=null, year=2018, volume=null, issue=null, pageStart=5, pageEnd=null, url=null, language=null, rfNumber=[27], rfOrder=48, authorNames=null, journalName=null, refType=null, unstructuredReference=SF/Z JD0107025-2018 Liquid Chromatography-Tandem Mass Spectrometry Method for the Detection of 15 Drugs and Its Metabolite in Hair[S]. 2018:5, articleTitle=null, 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AuthorCompanyExt(id=1239250706939572630, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, companyId=1239250706926989716, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.湖南警察学院侦查系,长沙 410138)]), AuthorCompany(id=1239250707036041628, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, xref=2., ext=[AuthorCompanyExt(id=1239250707044430238, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, companyId=1239250707036041628, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.Narcotics Detachment, Changsha Police Security Bureau, Changsha 410007, China), AuthorCompanyExt(id=1239250707052818848, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, companyId=1239250707036041628, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.长沙市公安局禁毒支队,长沙 410007)])], figs=[ArticleFig(id=1239250711725273755, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=EN, label=Fig.1, caption=MRM chromatograms of blank hair added with SKF525A(A),dextromethorphan(B) and dextrorphan(C), figureFileSmall=DdkH+CM2bYIH3Qq6OFBxFg==, figureFileBig=5AnagCSbVkGsbyfYv7PrEA==, tableContent=null), ArticleFig(id=1239250711842714274, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=CN, label=图1, caption=空白毛发提取液添加SKF525A(A)、右美沙芬(B)、去甲右美沙芬(C) MRM的色谱图, figureFileSmall=DdkH+CM2bYIH3Qq6OFBxFg==, figureFileBig=5AnagCSbVkGsbyfYv7PrEA==, tableContent=null), ArticleFig(id=1239250712065012402, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=EN, label=Tab.1, caption=

Linear gradient elution procedure

, figureFileSmall=null, figureFileBig=null, tableContent=
时间
(time)/min
流速
(flow rate)/(mL·min-1)
流动相比例(mobile phase ratio)/%
流动相
(mobile phase) A
流动相
(mobile phase) B
00.395.05.0
4.000.320.080.0
8.000.35.095.0
9.000.395.05.0
10.000.395.05.0
), ArticleFig(id=1239250712157287099, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=CN, label=表1, caption=

线性洗脱程序

, figureFileSmall=null, figureFileBig=null, tableContent=
时间
(time)/min
流速
(flow rate)/(mL·min-1)
流动相比例(mobile phase ratio)/%
流动相
(mobile phase) A
流动相
(mobile phase) B
00.395.05.0
4.000.320.080.0
8.000.35.095.0
9.000.395.05.0
10.000.395.05.0
), ArticleFig(id=1239250712249561791, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=EN, label=Tab.2, caption=

Qualitative ion pair,collision energy and retention time of SKF525A,dextromethorphan and dextrorphan

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
定性离子对
(qualitative ion pair) m/z
裂解电压
(fragmentor)/V
碰撞能量
(collision energy)/eV
保留时间
(retention time)/min
SKF525A354.2/209.245216.55
354.2/167.130
右美沙芬(dextromethorphan)272.3/213.360375.75
272.3/147.242
去甲右美沙芬(dextrorphan)258/15760455.18
258/13355
), ArticleFig(id=1239250712362808004, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=CN, label=表2, caption=

SKF525A、右美沙芬和去甲右美沙芬的定性离子对、裂解电压、碰撞能量和保留时间

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
定性离子对
(qualitative ion pair) m/z
裂解电压
(fragmentor)/V
碰撞能量
(collision energy)/eV
保留时间
(retention time)/min
SKF525A354.2/209.245216.55
354.2/167.130
右美沙芬(dextromethorphan)272.3/213.360375.75
272.3/147.242
去甲右美沙芬(dextrorphan)258/15760455.18
258/13355
), ArticleFig(id=1239250712467665611, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=EN, label=Tab.3, caption=

Recoveries and precisions

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
添加量
(added)/(ng·mg-1)
回收率
(recovery)/%
日内精密度
(intra-day RSD)/%
日间精密度
(inter-day RSD)/%
右美沙芬(dextromethorphan)0.198.41.53.3
0.597.61.52.1
2101.01.62.4
去甲右美沙芬(dextrorphan)0.1101.73.25.5
0.597.03.02.4
2104.83.92.7
), ArticleFig(id=1239250713965032148, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=CN, label=表3, caption=

回收率及精密度(n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
添加量
(added)/(ng·mg-1)
回收率
(recovery)/%
日内精密度
(intra-day RSD)/%
日间精密度
(inter-day RSD)/%
右美沙芬(dextromethorphan)0.198.41.53.3
0.597.61.52.1
2101.01.62.4
去甲右美沙芬(dextrorphan)0.1101.73.25.5
0.597.03.02.4
2104.83.92.7
), ArticleFig(id=1239250714074084055, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=EN, label=Tab.4, caption=

Contents of dextromethorphan and dextrorphanin in hair samples

, figureFileSmall=null, figureFileBig=null, tableContent=
序号
(No.)
右美沙芬(dextromethorphan)去甲右美沙芬(dextrorphan)
含量(content)/(ng·mg-1)RSD/%含量(content)/(ng·mg-1)RSD/%
K100
T12.111.40.451.0
T26.461.20.282.4
T35.862.20.032.0
T40.271.90.042.6
T57.152.70.722.2
T63.961.90.420.59
), ArticleFig(id=1239250714191524574, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239250703600906490, language=CN, label=表4, caption=

滥用者毛发样品中右美沙芬、去甲右美沙芬含量(n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
序号
(No.)
右美沙芬(dextromethorphan)去甲右美沙芬(dextrorphan)
含量(content)/(ng·mg-1)RSD/%含量(content)/(ng·mg-1)RSD/%
K100
T12.111.40.451.0
T26.461.20.282.4
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UPLC-MS/MS法检测人体毛发中右美沙芬及其代谢物去甲右美沙芬*
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赵楠 1 , 周国梁 2 , 黎书和 2 , 张旭东 2 , 田元 1 , 关力畅 1, **
药物分析杂志 | 代谢分析 2024,44(4): 603-609
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药物分析杂志 | 代谢分析 2024, 44(4): 603-609
UPLC-MS/MS法检测人体毛发中右美沙芬及其代谢物去甲右美沙芬*
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赵楠1 , 周国梁2, 黎书和2, 张旭东2, 田元1, 关力畅1, **
作者信息
  • 1.湖南警察学院侦查系,长沙 410138
  • 2.长沙市公安局禁毒支队,长沙 410007
  • Tel:15626259377;E-mail:

通讯作者:

** Tel:18274898856;E-mail:
Detection of dextromethorphan and dextrorphan in hair by UPLC-MS/MS*
Nan ZHAO1 , Guo-liang ZHOU2, Shu-he LI2, Xu-dong ZHANG2, Yuan TIAN1, Li-chang GUAN1, **
Affiliations
  • 1.Investigation Department, Hunan Police Academy, Changsha 410138, China
  • 2.Narcotics Detachment, Changsha Police Security Bureau, Changsha 410007, China
出版时间: 2024-04-30 doi: 10.16155/j.0254-1793.2024.04.07
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目的:

建立一种准确、快速检测毛发中右美沙芬及其代谢物去甲右美沙芬的超高效液相色谱-串联质谱(UPLC-MS/MS)方法。

方法:

用含内标双苯戊二氨酯(SKF525A)的甲醇溶液提取含有右美沙芬及其代谢物的毛发,经0.22 μm微孔有机滤膜过滤,UPLC-MS/MS检测。采用ACQUITY UPLC HSS T3超高液相色谱柱(100 mm×2.1 mm,1.8 μm),以0.2%甲酸(10 mmol·L-1甲酸铵)和乙腈为流动相进行梯度洗脱,流速0.3 mL·min-1,柱温为室温。采用电喷雾离子源,正离子多反应模式检测。

结果:

右美沙芬和去甲右美沙芬质量浓度均在1~100 ng·mL-1范围内线性响应良好,线性方程分别为Y=1.349 49X-0.020 80 (r=0.998 8)和Y=0.775 10X-0.013 87 (r=0.999 1),检测限和定量限均分别为0.01 ng·mL-1和0.025 ng·mL-1,平均回收率在97.0%~104.8%,日内精密度和日间精密度分别在1.5%~3.9%和2.1%~5.5%。将本方法应用于实际案例,结果在6例右美沙芬滥用者毛发中均检测出右美沙芬及其代谢物去甲右美沙芬。

结论:

该方法操作简便,检测灵敏度高,可用于毛发中右美沙芬及其代谢物去甲右美沙芬的检测。

超高效液相色谱-串联质谱(UPLC-MS/MS)  /  毛发  /  右美沙芬  /  去甲右美沙芬  /  法医毒物
Objective:

To establish a rapid and accurate ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the detection of dextromethorphan and one metabolite dextrorphan in hair.

Methods:

The hair sample containing dextromethorphan and dextrorphan were extracted with methanol containing internal standard proadifen hydrochloride(SKF525A). The extract was filtered with 0.22 μm organic filter membrane and detected by UPLC-MS/MS. All components were separated by an ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm), using a gradient elution procedure consisting of 0.2% formic acid (10 mmol·L-1 ammonium formate) and acetonitrile, at a flow rate of 0.3 mL·min-1, and the column temperature was room temperature. Positive electrospray ionization was performed using multiple reaction monitoring mode (MRM).

Results:

The linear relationships of dextromethorphan and dextrorphan were good in the range of 1-100 ng·mL-1. The linear equations were Y=1.349 49X-0.020 80 (r=0.998 8) and Y=0.775 10X-0.013 87 (r=0.999 1), respectively. The detection limit and quantitation limit were 0.010 ng·mL-1 and 0.025 ng·mL-1 respectively. Their recoveries ranged from 97.0%-104.8%. The intra-day RSD and inter-day RSD were 1.5%-3.9% and 2.1%-5.5%, respectively. The method was applied to cases, and the results showed that dextromethorphan and dextrorphan were detected in the hair of 6 abusers.

Conclusion:

This method is simple and sensitive enough to be applied to detect dextromethorphan and one metabolite dextrorphan in hair.

ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)  /  hair  /  dextromethorphan  /  dextrorphan  /  forensic toxicology
赵楠, 周国梁, 黎书和, 张旭东, 田元, 关力畅. UPLC-MS/MS法检测人体毛发中右美沙芬及其代谢物去甲右美沙芬*. 药物分析杂志, 2024 , 44 (4) : 603 -609 . DOI: 10.16155/j.0254-1793.2024.04.07
Nan ZHAO, Guo-liang ZHOU, Shu-he LI, Xu-dong ZHANG, Yuan TIAN, Li-chang GUAN. Detection of dextromethorphan and dextrorphan in hair by UPLC-MS/MS*[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (4) : 603 -609 . DOI: 10.16155/j.0254-1793.2024.04.07
右美沙芬(dextromethorphan,又名美沙芬)作用于脑干的咳嗽中枢,被作为止咳药物广泛使用[1]。右美沙芬是非竞争性的N-甲基-D-天冬氨酸(N-methyl-D-aspartate,NMDA)受体拮抗剂[2]。超剂量下可能产生与其他NMDA受体拮抗剂(苯环利定和氯胺酮)相似的临床症状,如欣快感和解离性幻觉,对周遭一切人事物极度满意、精神兴奋等[2-3],过量服用则会造成毒性反应[4],右美沙芬在人体内由细胞色素P450 2D6 (CYP2D6)酶代谢,生成去甲右美沙芬(dextrorphan)[2]
近年来,国内外报道了多起右美沙芬滥用及成瘾性事件[5-8],青少年群体中滥用右美沙芬来寻求精神放松的现象屡见不鲜[9]。滥用右美沙芬会出现兴奋、幻觉、轻快感、头晕等症状,长期超剂量服用则引起焦虑抑郁、暴躁冲动、幻觉妄想等精神障碍,产生心理依赖,耐药性增加,停药后出现戒断反应[10-12]。2021年底,由于滥用问题严重,国家药品监督管理局将右美沙芬由非处方药转为处方药。其左旋对映体左美沙芬已被列入麻醉药品目录。但由于右美沙芬仍未被我国列为管制类麻醉药品,常被作为毒品替代物进行吸食。
目前,关于右美沙芬及其代谢物去甲右美沙芬的检测方法较为成熟,常见的有高效液相色谱法(HPLC法)[13-15]和液相色谱串联质谱法(LC-MS/MS法)[16-18],多用于尿液及血浆等生物检材中。葛庆华等[19]与林凌云等[20]分别采用HPLC法检测血浆与尿液中的右美沙芬及去甲右美沙芬,然而其定量限较高(1~20 ng·mL-1)。徐丽云等[16]与刘丹等[21]采用LC-MS/MS法分别检测血液中的右美沙芬及去甲右美沙芬,定量限分别为0.98、0.2 ng·mL-1,与HPLC法相比更为灵敏,然而其回收率较低(87.16%±5.29%、70%~75%)。毛发因具有采集方便,易保存,覆盖用药的时限较长等优势,被广泛应用于违禁药品检测及毒情监测[22-23]。而目前尚缺乏关对于毛发中右美沙芬及其代谢物去甲右美沙芬检测技术的研究。
本研究建立了操作便捷,准确性较高的UPLC-MS/MS检测方法,用于检测毛发中右美沙芬及其代谢物去甲右美沙芬的含量。该方法结果稳定可靠,检出限较低可为今后司法实践中涉及右美沙芬滥用案件的处理提供有益参考。
高效液相色谱-串联质谱系统:ACQUITY UPLC HSS T3超高效液相色谱仪(Waters公司)串联SCIE QTRAP 6500质谱仪(AB SCIEX公司);EYELA高速振动器,上海爱朗仪器有限公司;AUW220D电子天平电子天平(精度0.01 mg),岛津公司;DAFT-01研磨仪,上海净信实业发展有限公司;F1-ClipTip可变容量单通道移液器,ThermoFisher公司。
右美沙芬标准溶液(1 mg·mL-1),来源于公安部三所,批号为FN11122001;去甲右美沙芬酒石酸盐(dextrorphantartrate,含量97.07%),购于TargetMol公司;盐酸双苯戊二氨酯(SKF525A,纯度95%),上海安谱实验科技公司。甲酸、乙腈(质谱纯)均购自麦克林试剂公司。空白毛发来自近期未服用过药物的志愿者,实验毛发由滥用右美沙芬人员自愿提供。
采用ACQUITY UPLC HSS T3超高效液相色谱柱(100 mm×2.1 mm,1.8 μm),以0.2%甲酸(10 mmol·L-1甲酸铵)为流动相A,乙腈为流动相B进行梯度洗脱(洗脱程序见表1),流速0.3 mL·min-1,进样量1 μL,柱温为室温。
离子源为点喷雾离子源,正离子模式(ESI+);离子源温度为550 ℃,离子喷雾电压(IS)为5 500 V;碰撞气(CAD)、气帘气(CUR)、雾化气(GS1)、辅助气(GS2)均为高纯氮气,使用前调节各气流流量以使质谱灵敏度达到检测要求;喷雾电压(IS)、去簇电压(DP)、碰撞能量(CE)等电压值优化至最佳灵敏度;检测方式为多反应检测模式(MRM)。每个化合物分别选择2对母离子/子离子对作为定性离子对,其定性离子对、定量离子对、裂解电压、碰撞能量和保留时间见表2
取毛发20 mg,加入甲醇1 800 μL,100 ng·mL-1 SKF525A标准溶液(取SKF525A 10 mg,溶解至甲醇9.5 mL中,再用甲醇稀释10 000倍,即得)200 μL,经研磨仪研磨5 min,频率为70 Hz,取研磨液经0.22 μm微孔有机滤膜过滤至1.5 mL进样瓶中,得到定性检验溶液,备检。
取甲醇2.8 mL,加入去甲右美沙芬酒石酸盐4.56 mg(去甲右美沙芬质量为2.8 mg),溶解,混合均匀后即得去甲右美沙芬标准溶液。分别取右美沙芬标准溶液、去甲右美沙芬标准溶液各100 μL,加入甲醇800 μL,漩涡混匀,得到0.1 mg·mL-1的混合标准溶液,置4 ℃冰箱待用。
取混合标准溶液200 μL,用甲醇定容至100 mL,制成右美沙芬、去甲右美沙芬质量浓度均为200 ng·mL-1的混合标准工作液。取空白毛发20 mg,加入混合标准工作液100 μL,按照“2.2”项方法操作处理检材,制成右美沙芬及去甲右美沙芬质量浓度均为10 ng·mL-1的溶液。按照“2.1”项下条件进行UPLC-MS/MS检测,在该色谱-质谱条件下,右美沙芬、去甲右美沙芬均得到分离,特异性良好,检材内源性杂质对检测目标物无干扰。SKF525A、右美沙芬、去甲右美沙芬出峰时间分别为6.55、5.75和5.18 min。色谱图见图1
取空白毛发6份,每份20 mg,分别加入右美沙芬、去甲右美沙芬混合标准工作液10、20、100、200、500、1 000 μL,按照“2.2”项方法操作,分别制成右美沙芬及去甲右美沙芬质量浓度均分别为1、2、10、20、50、100 ng·mL-1,SKF525A质量浓度均为10 ng·mL-1的溶液,总体积为2 mL。按照“2.1”项下条件进行HPLC-MS/MS检测,以右美沙芬、去甲右美沙芬色谱峰峰面积之比(Y)为纵坐标,右美沙芬、去甲右美沙芬浓度(X,ng·mL-1)为横坐标,进行线性回归。线性方程分别为
右美沙芬、去甲右美沙芬质量浓度在0.1~100 ng·mL-1范围内线性关系良好。
将检测限定为色谱峰峰高相当于基线噪音3倍时的浓度,将定量限定为色谱峰峰高相当于基线噪音10倍时的浓度。右美沙芬与去甲右美沙芬的检测限均为0.01 ng·mL-1,定量限均为0.025 ng·mL-1
取空白毛发,每份20 mg,分别加入右美沙芬、去甲右美沙芬混合标准工作液10、50、200 μL,按“2.2”项下方法制备毛发中右美沙芬及去甲右美沙芬含量分别为0.1、0.5、2 ng·mg-1的待测溶液各6份,按“2.1”项的条件进行UPLC-MS/MS检测;根据回归方程计算样品中右美沙芬及去甲右美沙芬浓度。以测得溶液中右美沙芬及去甲右美沙芬浓度与添加浓度之比来计算回收率。以不同浓度溶液同日内重复测定6次计算日内精密度,不同浓度溶液6日内重复测定6次计算日间精密度,结果见表3。右美沙芬与去甲右美沙芬低、中、高3个浓度的平均回收率分别为97.6%~101.0%和97.0%~104.8%,日内精密度和日间精密度分别为1.5%~3.9%和2.1%~5.5%。试验结果均符合2020年版《中华人民共和国药典》分析方法验证指导原则中关于回收率与精密度的要求[24]
6例滥用右美沙芬人员编号为T1、T2、T3、T4、T5、T6,分别取毛发3份,每份20 mg。取空白毛发1份,作为对照,编号为K1。按照“2.2”项下方法进行处理,按“2.1”项的条件进行UPLC-MS/MS检测。滥用右美沙芬人员的毛发样品中右美沙芬、去甲右美沙芬出现的色谱峰保留时间与“2.4.2”项中的色谱峰保留时间的相对误差分别为0.7%、1.36%,且特征碎片离子均出现。右美沙芬、去甲右美沙芬离子相对丰度比与“2.4.2”项中的离子相对丰度比的相对误差分别为12.5%、4.28%,符合中华人民共和国司法部鉴定管理局发布的司法鉴定技术规范要求[25-27]。综上可判断检测的毛发样品中存在右美沙芬及去甲右美沙芬化合物。滥用右美沙芬人员的毛发样品中右美沙芬及去甲右美沙芬含量检测结果见表4,空白对照组中未检出待测成分。滥用者毛发样品右美沙芬含量检测的精密度为1.2%~2.7%,去甲右美沙芬含量的检测精密度为0.59%~2.6%。
本研究在“毛发中15种毒品及代谢物的液相色谱-串联质谱检验方法”等司法鉴定技术规范基础上不断优化试验条件与方法,建立UPLC-MS/MS检测毛发中右美沙芬及去甲右美沙芬的方法。在色谱条件方面,本文考察了ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8 μm)、ACQUITY UPLC CSHTM C18(100 mm×2.1 mm,5 μm)2种色谱柱;在流动相方面,本文对比考察了甲醇、乙腈2种有机相,在甲酸水溶液中加入不同浓度的甲酸铵;流动相比例则考察了等度和梯度洗脱,测试了不同的流速,最终选择ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8 μm)色谱柱,0.2%甲酸(10 mmol·L-1甲酸铵)-乙腈流动相组合,梯度洗脱,0.3 mL·min-1流速等色谱条件,可以获得最佳色谱保留及峰形。在检材处理方面,本文对毛发研磨的频率、时间均进行了考察,最终确定频率为70 Hz,时间为5 min,可最大效率提取毛发中的目标成分。
与其他基于血液、尿液等生物检材的右美沙芬及去甲右美沙芬检测方法[1619-21]相比,本文建立了人体毛发中右美沙芬和去甲右美沙芬的UPLC-MS/MS检测方法。相较于其他生物检材,毛发中药物及其代谢物残留时间更长,通常作为一种重要的补充手段在司法实践中应用广泛。本研究中右美沙芬及去甲右美沙芬的检测限均为0.01 ng·mL-1,定量限均为0.025 ng·mL-1,回收率分别为101%~125%和95%~115%。相比较于已报道的HPLC法[19-20]和LC-MS/MS法[1621],检测限与定量限更低,稳定性更高,具有显著优势。
本方法对6例右美沙芬滥用者进行了毛发检测,得到了可靠验证。然而,从表4的实验结果可以看出,滥用者毛发中右美沙芬含量与其代谢物去甲右美沙芬含量并无明显的相关关系。例如:编号T5毛发中的右美沙芬含量最高,为7.15 ng·mg-1,其代谢物去甲右美沙芬含量也为最高0.72 ng·mg-1。然而编号T4毛发中的右美沙芬含量最低,为0.27 ng·mg-1,但是其代谢物去甲右美沙芬含量为0.04 ng·mg-1,并非最低。笔者认为这可能与个体吸食时间、代谢差异等因素有关。因此,具体的代谢规律有待进一步研究。此外,本研究也对医疗用途服用右美沙芬的患者毛发进行了检测,结果显示其毛发中的右美沙芬含量远低于滥用人员毛发中的含量。
综上,本研究成功建立了一种毛发中右美沙芬及其代谢物去甲右美沙芬的UPLC-MS/MS分析方法,操作简便快捷,检测结果稳定,准确性好,较之其他方法检出限更低,更为灵敏,可作为一种可靠方法应用于司法实践中毛发右美沙芬及其代谢物去甲右美沙芬的检测。
  • *湖南省社会科学成果评审委基金项目(XPS2023JYC034)
  • 湖南警察学院新型犯罪案件侦查理论与实践科研创新团队
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doi: 10.16155/j.0254-1793.2024.04.07
  • 接收时间:2023-06-23
  • 首发时间:2026-03-13
  • 出版时间:2024-04-30
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  • 收稿日期:2023-06-23
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*湖南省社会科学成果评审委基金项目(XPS2023JYC034)
湖南警察学院新型犯罪案件侦查理论与实践科研创新团队
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    1.湖南警察学院侦查系,长沙 410138
    2.长沙市公安局禁毒支队,长沙 410007

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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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