阿片类药物的检测

阿片类物质主要包括吗啡、海洛因、可待因等阿片类生物碱,是常见的一类滥用药物.本文就近些年来对阿片类物质及代谢产物检测方法的研究作一介绍.     

阿片类成瘾者尿中吗啡的测定及代谢产物的鉴定

建立了阿片类生物碱系统的气相色谱一质谱分离分析方法。在所选色谱条件下八种阿片类生物碱均能得到良好的分离，并用所建方法对阿片类成瘾者尿样进行了吗啡的测定及代谢产物的鉴定，尿中检测出吗啡、可待因、６－单乙酰基吗啡及微量的乙酰可待因，从而确定该吸毒者吸食的毒品为海洛因粗提物。     

液相色谱-串联质谱法测定头发中11种阿片类生物碱

建立头发中海洛因、吗啡、单乙酰吗啡等11种阿片类生物碱的液相色谱-串联质谱测定方法,并考察海洛因滥用者头发中阿片类组分的存在情况。头发经冷冻研磨后加入硼酸缓冲液超声30 min,用氯仿-异丙醇(9∶1)提取。用Allure PFP丙基柱,以乙腈-乙酸铵(0.1%甲酸)梯度洗脱分离,采用二级质谱多反应监测模式(MRM)检测11种阿片类生物碱。头发中海洛因、吗啡、单乙酰吗啡等11种阿片类生物碱在对应质量浓度范围内线性良好(r>0.996 0);检测限(LOD)均小于0.05 ng.mg-1;回收率范围为47.2%～110%;日内精密度和日间精密度均小于14%。21例海洛因滥用者头发中均检出了海洛因、单乙酰吗啡、吗啡、可待因、乙酰可待因、氢可酮等主要组分。所建方法灵敏度高、选择性好,适用于同时分析头发中海洛因等11种阿片类生物碱组分,可有效鉴别海洛因滥用与阿片类药物或食品的摄取。     

LC/APCI/MS在血药浓度研究中的应用

目的：建立可用于血样检测的小分子药物代谢分析的高灵敏、简捷的液相色谱/大气压解离质谱联用(LC/APCI／MS)方法。方法：海洛因、6—乙酰吗啡、乙酰可待因和螺内酯分别于大鼠血浆中39℃孵育，定时取样，用LC/APCI／MS测定原药及代谢产物的浓度，绘制代谢曲线。结果：海洛因在5min内消失，经6—乙酞吗啡最终代谢为吗啡，30min后吗啡浓度达到峰值；6—乙酰吗啡在20min内消失，代谢产物吗啡的浓度随之达到峰值；乙酞可待因在血浆中代谢缓慢，60min后仅有部分代谢，生成可待因；螺内酯也以缓慢的速度代谢，60min后代谢产物坎利酮的浓度尚远离峰值。结论：LC/APCI／MS作为高灵敏废的简捷方法用于血药浓度研究具有广阔前景。     

生物检材中的常见滥用物质分析及比较

1常见滥用物质的分类及其相关代谢产物常见的滥用物质主要为麻醉药品和精神药品两大类。常见的麻醉药品包括阿片、吗啡、海洛因、可卡因、大麻等。常见的精神药品包括镇静催眠药(如苯二氮类、巴比妥类等)、苯丙胺、甲基苯丙胺(俗称“冰毒”)、3、4-二亚甲基二氧安非他明(MDMA,俗称“摇头丸”)、麦角酰二乙胺(LSD)等。其他滥用物质有烟草、酒精、挥发性有机溶剂艹卓吗啡分为精制吗啡、粗制吗啡。精制吗啡一般用于医疗用途。被滥用的吗啡一般为粗制吗啡。粗制吗啡是从阿片膏提取的,主要含有吗啡,也含有可待因、蒂巴因、罂粟碱和那可丁。精…     

零包海洛因的成分、含量以及海洛因滥用者尿液中吗啡/可待因比值

目的:探讨零包海洛因的成分、含量及海洛因滥用者尿液中吗啡/可待因比值。方法:采用气相色谱/质谱联用法(GC/MS)和气相色谱法(GC)检测105份海洛因滥用者的尿样及441件零包海洛因,并作描述性统计分析。结果:105份尿样中有9份检出海洛因原体(其中三份同时检出O6-单乙酰吗啡);30份仅检出吗啡和可待因,其中吗啡/可待因峰面积比值小于1的有13份。441件零包中海洛因含量的最大值是95.2%,最小值为0;乙酰可待因含量的最大值是89.3%,最小值是1.6%。乙酰可待因含量比海洛因含量大的零包有25例,占到5.7%。结论:零包中海洛因中的成分及含量差别大;嫌疑人的尿液中吗啡/可待因浓度比小于1时,不能排除有吸食海洛因的可能。     

尿液LC-MS/MS检测方法快速甄别海洛因滥用者

目的獉獉:建立快速甄别海洛因滥用者的液相色谱-串级质谱法(LC-MS/MS)定性方法。方法獉獉:采用LC-MS/MS检测两个试验组即服用复方甘草片组和服用可愈糖浆后于不同时间点留取的尿样及162例海洛因滥用者的尿样,并进行描述性统计分析。结果獉獉:两个试验组尿液中均检测出吗啡、可待因、3-β-D–葡萄糖醛酸吗啡(M3G)和6-葡萄糖醛酸可待因(C6G)等吗啡类药物成分,且约在0.5-3 h浓度达到高峰,但是尿液中总吗啡和总可待因比值(Mor/Cod)是不同的。服用复方甘草片组尿液总吗啡/总可待因峰面积比值(Mor/Cod):1.61±s 0.67≤Mor/Cod<3.07±s 1.85;服用可愈糖浆组0.09±s 0.07≤Mor/Cod<0.24±s 0.12;海洛因滥用者尿液中有的可测出6-MAM,有的还可检测出海洛因或乙酰可待因原形物质,162例尿样中9例尿样Mor/Cod比值小于1且9例均检测出6-MAM外,其余均大于1。结论獉獉:本方法用于快速甄别海洛因滥用与阿片类药物使用,是相对可行的。     

阿片类滥用对骨代谢的影响

吗啡、海洛因等阿片类药物的长期滥用可使个体产生强烈的精神依赖和躯体依赖，造成机体多系统功能的损害和改变。本文主要综述长期应用吗啡等阿片类物质对骨代谢影响。     

服用含罂粟壳药物尿液中阿片类物质的检测

部分止咳中成药物含有罂粟壳成分 ,罂粟壳进入人体后代谢产物的主要成分有可待因、吗啡、蒂巴因和罂粟碱等。其中的吗啡成分与吸食毒品海洛因后的最终代谢产物相同。吗啡胶体金检测技术主要应用于针对海洛因吸毒者的尿液中吗啡的检测 ,但也对服用止咳药物正常人产生阳性结果。罂粟壳和海洛因经人体代谢后的最终产物有差异 ,所以利用气相色谱技术检测罂粟壳代谢后的其它产物 ,可快速、准确地提供确实的客观证据。1材料与方法1 1实验材料盐酸吗啡标准样品 (公安部物证鉴定中心提供批号 :96 0 6 ) ;提取液为氯仿∶异丙醇 (3∶1) ;pH值为9 2的N…     

阿片滥用对甲状腺结构功能影响的研究进展

阿片类物质作用于阿片受体可以产生吗啡样作用,但因为其特殊的药理性质极易引起躯体、精神产生依赖并导致滥用。研究发现长期滥用阿片类物质会导致依赖者包括下丘脑-垂体-肾上腺、性腺、甲状腺轴在内的内分泌功能受到影响。阿片类物质对甲状腺结构、功能的影响尚未有系统的阐述,本文通过对阿片类物质作用下甲状腺形态结构、激素水平、甲状腺素结合蛋白等改变来阐述阿片对甲状腺的影响。     

Determination of morphine and codeine in urine by gas chromatography—mass spectrometry

GC—MS is one of the recommended analytical techniques for the identification and confirmation of opiates in urine. A method for the qualitative detection and quantitation of codeine and morphine in urine samples by this technique has been developed. This method is also suitable for the detection of their main metabolites in urine: norcodeine and normorphine. It also allows the identification of 6-monoacetylmorphine in urine, which can be used as a confirmatory marker of heroine abuse.

The derivatized compounds are separated by capillary gas chromatography (GC) and identified by mass spectrometry (MS) in the selective ion monitoring acquisition mode (SIM).

The recoveries from urine at concentrations of 1000 ng ml⁻¹ are 72% for codeine and 80% for morphine. The method is linear in the range studied (0–1000 ng ml⁻¹) for codeine and morphine.     

Comparison of the various opiate alkaloid contaminants and their metabolites found in illicit heroin with 6-monoacetyl morphine as indicators of heroin ingestion

In this study the use of the various opiate alkaloid contaminants as potential markers for illicit heroin ingestion were investigated. Urine samples (n = 227) taken from prisoners for routine drug screen, which were positive for opiates by immunoassay screening, were analyzed for contaminants in illicit heroin. A previously described method was used for the analysis; urines were extracted using mixed-mode solid-phase extraction; the extracts were derivatized using N-methyl-bistrifluoroacetamide and N-methyl-N-trimethylsilyltrifluoroactamide/trimethylchlorosilane. The derivatized extracts were subjected to electron impact gas chromatography-mass spectrometry. The extracts were injected in full scan mode followed by selected ion monitoring mode for target opiate alkaloids found as contaminants in illicit heroin. The opiate alkaloids and their metabolites specifically targeted included meconine, desmethylmeconine, hydrocotarnine, acetylcodeine, codeine, morphine, 6-monacetylmorphine (6-mam), papaverine, hydroxypapaverine, and dihydroxypapaverine. Of the 227 samples positive for opiates by immunoassay, using a cut-off of 300 ng/mL, 199 were confirmed positive for morphine and using a cut-off of 10 ng/mL, 28 were confirmed positive for 6-mam. Using the screening method described in the study, the following numbers of positives were found: 199 for morphine, 103 for codeine, 5 for meconine, 46 for desmethylmeconine, 18 for 6-mam, 136 for hydroxypapaverine, and 139 for dihydroxypapaverine. Acetylcodeine, hydrocotarnine, and papaverine were not detected in any of the samples. The results of this study show that analysis for papaverine metabolites is more sensitive than 6-mam as a way of demonstrating illicit heroin use.     

Confirmation of recent heroin abuse: Accepting the challenge

Confirmation or exclusion of recent heroin consumption is still one of the major challenges for forensic and clinical toxicologists. A great variety of biomarkers is available for heroin abuse confirmation, including various opium alkaloids (eg, morphine, codeine), street heroin impurities (eg, 6‐acetylcodeine [6‐AC], noscapine, papaverine) as well as associated metabolites (eg, 6‐monoacetylmorphine [6‐MAM], morphine glucuronides). However, the presence of most of these biomarkers cannot solely be attributed to a previous heroin administration but can, among other things, also be due to consumption of poppy seed products (‘poppy seed defense’), opium preparations or specific medications, respectively. A reliable allocation is of great importance in different contexts, for instance in the case of DUID (driving under the influence of drugs) investigations, in driving licence re‐granting processes, in workplace drug testing (WDT), as well as in post‐mortem identification of illicit opiate use. Additionally, differentiation between illicit street heroin abuse and pharmaceutical heroin administration is also important, especially within the frame of heroin‐assisted treatments. Therefore, analysis of multiple biomarkers is recommended when illicit opiate consumption is assumed to obtain the most reliable results possible. Beyond that, interpretation of positive opiate test results requires a profound insight into the great variety of biomarkers available and their validity regarding the alleged consumption. This paper aims to provide an overview of the wide variety of heroin abuse biomarkers described in the literature and to review them regarding their utility and reliability in daily routine analysis.     

Some aspects of the metabolism of morphine-N-oxide

After administration of morphine-N-oxide (MNO) to rats the opiates appearing in the urine were morphine (61%) and MNO (39%). After administration of morphine, the urinary opiates were morphine (80%) and normorphine (20%). When tacrine was given with morphine the urine also contained MNO (46% of total urinary opiates) and the amount of normorphine was much decreased (to 1%), the remainder being morphine (53%). Tacrine and amiphenazole inhibited demethylation of morphine and codeine by a rat liver fraction in vitro. MNO had weak inhibitory activity. Neither MNO nor codeine-N-oxide were demethylated in vitro.     

Simultaneous quantification of opiates and effect of pigmentation on its deposition in hair

In forensic toxicology, the abuse of various opiate preparations, such as raw opium and heroin, is of interest since the metabolic pathways of these opiates overlap. Although the pharmaco(toxico)kinetics in hair is not clearly understood, melanin is thought to play a key part in the incorporation and distribution of drugs and metabolites in hair. Therefore, in the present study, a simultaneous quantification method for the determination of codeine, morphine, norcodeine, normorphine and 6-acetylmorphine (6-AM) in hair was developed in order to analytically diagnose chronic users of opiates including morphine and codeine preparations, raw opium and heroin. Furthermore, the effect of hair pigmentation on the distribution of opiates in hair was investigated using lean Zucker rats with both dark grey and white hair on the same body. Opiates were extracted using 0.1 M hydrochloric acid followed by solid phase extraction. The extracts were derivatized with N-methyl-N-(trimethylsilyl)trifluoroacetamide and analyzed using gas chromatography/mass spectrometry (GC/MS). The method was fully validated and applied to the animal study. In conclusion, the current study demonstrates that codeine, morphine and their metabolites were successfully determined in both pigmented and non-pigmented hair. However, the melanin content plays an important role in the degree of incorporation of morphine, codeine and their metabolites into hair.     

Experience with a urine opiate screening and confirmation cutoff of 2000 ng/mL.

Until recently, most laboratories used an opiate immunoassay screening and confirmation cutoff value of 300 ng/mL for codeine and morphine detection by gas chromatography-mass spectrometry (GC-MS). The cutoff value for opiates was increased to 2000 ng/mL or higher in various laboratories because of concerns that small doses of codeine and foods containing poppy seeds would give a positive opiate-screening result. Workplace drug-testing programs in the U.S. raised the opiate cutoff value to 2000 ng/mL on 30 November 1998. The objective of this study is to describe the results of opiate testing of 8600 urine specimens collected over 24 months with a 2000-ng/mL screening and confirmation (codeine and morphine) cutoff value. Specimens were screened by the EMITdau opiate assay using an in-house 2000-ng/mL morphine calibrator. Presumptive positive findings (N = 621) were analyzed quantitatively by GC-MS for codeine and morphine. One hundred and eighty six urine specimens were positive for codeine and morphine (> 2000 ng/mL), 298 specimens were positive for codeine only (> 2000 ng/mL) and 26 specimens were positive for morphine only (> 2000 ng/mL). All remaining specimens had codeine and morphine values < 2000 ng/mL. The codeine and morphine confirmation rate in this program reduced from 7.1% in 1994-1996 (300-ng/mL cutoff) to 2.1% in 1997-1998 with a 2000-ng/mL cutoff value. The codeine-only confirmation rate lowered from 6.6% (300-ng/mL cutoff) to 3.4% (2000-ng/mL cutoff). It was concluded that increasing opiate screening and codeine and morphine confirmation cutoff values led to > 300% reduction in the confirmed-positive rate for codeine and morphine and a 47% reduction in codeine-only confirmations in a urine drug-testing program where codeine was the major opiate used.     

Analysis of morphine and codeine in samples adulterated with Stealth.

Stealth is an adulterant used to avoid detection of drug abuse. The product does have an effect on the ability to detect several drugs of abuse, including the opiates morphine and codeine. It has previously been shown that low concentration (2500 ng/mL morphine) samples adulterated with Stealth tested negative by both Roche OnLine and Microgenics CEDIA immunoassays, but those spiked with higher concentrations (6000 ng/mL of codeine and morphine glucuronide) were positive. Initial results showed confirmation analysis was also sometimes negatively impacted by this adulterant. Urine samples were spiked with 6000 ng/mL of codeine and/or morphine glucuronide to assess the effect of Stealth. Each individual sample was split into separate aliquots. One aliquot of each was adulterated with Stealth following package directions. The samples were then tested by immunoassay and gas chromatography-mass spectrometry (GC-MS). The control and adulterated aliquots were positive by both immunoassays. Results of GC-MS analysis of the Stealth-adulterated aliquots following standard procedures using deuterated internal standards proved unsuccessful in several cases. In 4 of 12 cases (33%), neither the drugs nor internal standards were recovered despite repeated attempts. In one other sample, recovery was dramatically reduced, making accurate quantitation impossible, whereas the unadulterated aliquots of the same samples posed no problem with recovery. Addition of sodium disulfite to the aliquots prior to extraction allowed recovery of the drugs and internal standards from all samples. Analysis of the samples showed the concentration of morphine and codeine decreased in some by as much as 17 and 30%, respectively. In other cases, there was essentially no difference in the concentration seen before and after adulteration, with or without disulfite treatment. Unless the initial concentration of opiate is near the cutoff, samples containing opiates are likely to be immunoassay positive, it is important to consider this procedure as an option for samples that screen positive but the opiates and their respective internal standards are not recovered for GC-MS analysis.