摘要
目的:建立同时测定毛发中6-单乙酰吗啡(6-AM)、吗啡(MOR)及可待因(COD)的高效液相色谱-串联质谱联用(HPLC-MS/MS)方法,并对吸毒者的毛发中3种海洛因代谢物浓度进行考察及阈值分析。方法:以6-AM-D3和MOR-D3为内标,毛发经甲醇常温超声后,采用RESTEK Ultra PFPP(2.1 mm×100 mm,5μm)色谱柱,流动相为乙腈-含0.1%(v/v)甲酸的20 mmol·L-1乙酸铵水溶液(70∶30, v/v),流速为0.3 mL·min-1;进样量为5μL。采用电喷雾离子源于正离子模式下进行多反应监测模式(MRM)进行检测。在该测定方法下,对188例海洛因吸毒者的毛发进行检测并对结果进行了统计。结果:6-AM、MOR及COD在定量范围内线性关系良好(r>0.999),检测下限分别0.002、0.004及0.002 ng·mg^-1,定量下限分别为0.008、0.01及0.008 ng·mg^-1,日内和日间精密度均<5%,低、中、高浓度回收率均>96%。将检出MOR作为金标准,绘制ROC曲线得出阈值为6-AM≥1 ng·mg^-1,曲线下面积为0.901,灵敏度74.4%,特异度87.3%;并以浓度大于检测下限为筛选条件,统计得出6-AM的均值为3.129 ng·mg^-1,最小值为0.056 ng·mg^-1,最大值为62.783 ng·mg^-1,中位数为1.506 ng·mg^-1,6-AM与MOR的比率的均值为2.503。结论:该检测方法灵敏度及专属性良好,可用于毛发中海洛因代谢物的检测。毛发中海洛因代谢物浓度结果受人种及发色的影响较大,当毛发中6-AM≥1 ng·mg^-1时,可判定该样本为海洛因长期使用情况。而6-AM及MOR的比率范围跨度较宽,可作为参考判断依据。
Objective:To establish an HPLC-MS/MS method for simultaneous determination of 6-acetylmorphie(6-AM),morphine(MOR)and codeine(COD)and to investigate cut-off of 6-AM,MOR and COD in heroin users’ hair.Methods:6-AM-D3 and MOR-D3 were used as internal standards.After the hair was sonicated in methanol at room temperature,the sample was separated on the RESTEK Ultra PFPP(2.1 mm×100 mm,5 μm)column.An injection volume of 5 μL were eluted with acetonitrile-20 mmol·L-1 ammonium acetate aqueous solution containing 0.1%(v/v)formic acid(70 ∶ 30,v/v)at 0.3 mL·min-1 flow rate,followed by detection in positive electrospray ionization by multiple reaction monitoring(MRM)mode.The concentration of 6-AM,MOR and COD from 188 heroin users were determined and analyzed.Results:The calibration curves of 6-AM,MOR and COD showed good linearity in the quantitative ranges(r>0.999).The limits of detection(LOD)were 0.002,0.004 and 0.002 ng·mg^-1,respectively,and the limits of quantification(LOQ)were 0.008,0.01 and 0.008 ng·mg^-1,respectively.Both intra-day and inter-day precision were less than 5%.Low,medium and high concentration recovery rates were higher than 96%.The cut-off value for 6-AM was found to be 1.01 ng·mg^-1 from the ROC curve for the simultaneous detection of MOR≥LOD.The AUC was 0.901,sensitivity and selectivity was 74.4% and 87.3%,respectively.Mean value of 6-AM was 3.129 ng·mg^-1, the minimum value was 0.056 ng·mg^-1,the maximum value was 62.783 ng·mg^-1,and the median was 1.506 ng·mg^-1.The mean ratio of 6-AM to MOR was 2.503.Conclusion:The method has good sensitivity and specificity and can be used for the detection of heroin metabolites in hair.The results of heroin metabolite concentration in hair are greatly affected by race and hair color.When the concentration of 6-AM in hair is higher than 1 ng·mg^-1,the sample was identified as heroin chronic user.The range of ratios of 6-AM and MOR is wide and can be used as judgement reference.
作者
张颖怡
欧军
陈深树
钟超群
杜娟
焦台风
徐代化
ZHANG Ying-yi;OU Jun;CHEN Shen-shu;ZHONG Chao-qun;DU Juan;JIAO Tai-feng;XU Dai-hua(Guangdong Nantian Institute of Forensic Science,Shenzhen 518003,China;Department of Hygiene Inspection and Quarantine Science,School of Public Health,Southern Medical University,Guangzhou 510515,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2020年第12期2157-2166,共10页
Chinese Journal of Pharmaceutical Analysis
基金
广东省深圳市科技创新委员会2017年技术创新计划技术攻关项目(JSGG2017041310065739)
广东省医学科研基金项目(A2018229)。
