新型微流体液相色谱(micro-LC)串联四极杆质谱检测头发中5种苯二氮卓类药物

目的建立了新型微流体液相色谱(micro-LC)串联四级杆质谱检测头发中5种苯二氮卓类药物(三唑仑,α-羟基咪达唑仑,咪达唑仑,α-羟基阿普唑仑,普拉西泮)。方法采用液-液萃取法(LLE)提取毛发样品,微流体液相色谱串联四级杆质谱检测。结果在回归系数大于0.99的情况下,目标分析物的良好线性。精密度和萃取效率分别在2.7%~17.1%和66.5%~120.5%之间。每个分析物的检出限和定量限分别为0.001~0.08 pg·mg^-1和0.0125~0.25pg·mg^-1。结论本方法与传统的液相色谱串联质谱法(LC/MS-MS)相比灵敏度出高2-10倍。本研究展示了新型微流体分离系统串联质谱方法的实用性,并为法医毒理学领域关于头发样本中痕量药物检测提供了一种新的检测手段。     

固相萃取／LC-MS／MS测定尿液中吗啡类药物

目的 建立尿液中吗啡类药物的固相萃取／LC—MS／MS方法。方法采用OASIS MCX3cc（60mg）固相萃取柱进行提取,应用LC—MS／MS方法进行检测,运用保留时间和MRM方式对尿液中吗啡类药物及其代谢物进行定性定量分析。结果磷酸盐缓冲液pH4．0时,海洛因、6-MAM、可待因、吗啡、M3G的固相萃取回收率分别达64．33％-70．21％,96．95％～117．57％,83．60％～123．63％,68．82％～91．03％,94．64％～107．33％;最低检测限（LOD）分别为5、10、5、5、2pg,线性范围0．005～10μg／mL;相关系数分别为0．9998、0．9958、0．9992、0．9994、0．9997。结论本文所建方法,适用于尿液中吗啡类药物的分析。     

A novel method for the analysis of discharged smokeless powder residues

A novel method for the estimation of intermediate-long firing distance range is proposed. The method is based on the characterization and chemical analysis of the smokeless powder particles on the target. An adhesive lifter is applied to collect the suspected gunshot residues (GSRs) from the surface of an object, and a Modified Griess Test (MGT) is carried out after alkaline hydrolysis on the adhesive lifter. Visualized particles are removed from the adhesive lifter under a microscope. Two systems are used for the analysis of organic discharge residues from the smokeless powder: (1) gas chromatography/thermal energy analysis (GC/TEA) for the analysis of nitroglycerine (NG) and 2,4-dinitrotoluene (2,4-DNT), (2) gas chromatography/mass spectrometry (GC/MS) for the identification of organic components such as DNT, NG, and some stabilizers. By using this procedure and confirming that the suspected particles are indeed GSR, one can estimate the intermediate-long firing distance of c. 0.75-3 m in the presence of very few particles and provide information for the classification of ammunition type in casework.     

生物检材中苯丙胺类兴奋剂和氯胺酮的LC-MS/MS分析

目的建立生物检材中苯丙胺类兴奋剂和氯胺酮的液相色谱-串联质谱(LC-MS/MS)分析方法。方法生物检材包括血液、尿液和毛发,采用稀释法和液液提取的前处理方法,应用两个不同的液相柱,优化LC-MS/MS分析方法,并考察了血液和尿液基质的离子抑制作用。结果同时分析苯丙胺和MDA,液相1在3m in内完成,液相2可用于确认分析或复杂基质分离。尿液稀释法检材用量少,前处理简便快速。毛发中苯丙胺类兴奋剂和氯胺酮的最低检测限(LOD)为0.005~0.05ng/mg。对送检案例检材产妇头发和胎毛进行苯丙胺类兴奋剂和氯胺酮的分析。结论本方法可用于生物检材中苯丙胺类兴奋剂和氯胺酮的同时分析,血、尿等生物检材的离子抑制作用是影响本方法灵敏度的主要原因。     

固相萃取/LC-MS/MS测定血液中的白藜芦醇

目的建立血液中白藜芦醇的固相萃取/LC-MS/MS方法。方法自制固相萃取柱,在长约13cm的酸性滴定管中,底部填入约0.02g玻璃棉,依次加入柱填料PSA 0.15g,Florisil 0.3g,Silica 0.45g。对样本进行固相萃取,采用LC-MS/MS方法进行检测,运用保留时间和多反应监测(MRM)方式对血液中白藜芦醇进行定性定量分析。结果回收率为95.66%～98.58%,最低检测限(LOD)为1.79μg/mL,线性范围2.5～20μg/mL;相关系数为0.9997。结论本文所建方法,适用于血液中白藜芦醇的检测。     

Investigating the effect of changing ammunition on the composition of organic additives in gunshot residue (OGSR)

The measurement of the organic additives in smokeless gunpowder is an attractive approach for the detection of handgun use because it provides compositional information that can help associate residues and unfired gunpowder. We investigate several factors that will be required to advance the characterization of organic gunshot residue (OGSR) as a useful forensic tool, including evaluating residue contamination from previously fired ammunition, particle-to-particle compositional variability, and compositional features resulting from the type of firing primer used. Using ammunition loaded with known smokeless powders containing different stabilizers, a sequence of shots was fired from a .357 magnum revolver, and the muzzle exit residues were collected. Compositional analysis of the residues, both in bulk and as single particles, showed only a trace of the previously fired powder in the first shot and none in subsequent shots. In an additional experiment testing conventional leaded and the new lead-free firing primers, the OGSR composition was found not to depend on the primer type.     

Quantitative analysis of multiple illicit drugs in preserved oral fluid by solid-phase extraction and liquid chromatography–tandem mass spectrometry

We present a validated method for the simultaneous analysis of basic drugs which comprises a sample clean-up step, using mixed-mode solid-phase extraction (SPE), followed by LC–MS/MS analysis. Deuterated analogues for all of the analytes of interest were used for quantitation. The applied HPLC gradient ensured the elution of all the drugs examined within 14 min and produced chromatographic peaks of acceptable symmetry. Selectivity of the method was achieved by a combination of retention time, and two precursor-product ion transitions for the non-deuterated analogues. Oral fluid was collected with the Intercept^®, a FDA approved sampling device that is used on a large scale in the US for workplace drug testing. However, this collection system contains some ingredients (stabilizers and preservatives) that can cause substantial interferences, e.g. ion suppression or enhancement during LC–MS/MS analysis, in the absence of suitable sample pre-treatment. The use of the SPE was demonstrated to be highly effective and led to significant decreases in the interferences. Extraction was found to be both reproducible and efficient with recoveries >76% for all of the analytes. Furthermore, the processed samples were demonstrated to be stable for 48 h, except for cocaine and benzoylecgonine, where a slight negative trend was observed, but did not compromise the quantitation. In all cases the method was linear over the range investigated (2–200 μg/L) with an excellent intra-assay and inter-assay precision (coefficients of variation <10% in most cases) for QC samples spiked at a concentration of 4, 12 and 100 μg/L. Limits of quantitation were estimated to be at 2 μg/L with limits of detection ranging from 0.2 to 0.5 μg/L, which meets the requirements of SAMHSA for oral fluid testing in the workplace. The method was subsequently applied to the analysis of Intercept^® samples collected at the roadside by the police, and to determine MDMA and MDA levels in oral fluid samples from a controlled study.     

液相色谱-串联质谱法检测血液中的美西律

目的建立测定血液中美西律(mexiletine)的液相色谱-串联质谱联用法(LC-MS/MS)。方法采用简便的乙腈蛋白沉淀法对血液进行预处理,应用Allure PFP Propyl液相柱分离,用电喷雾正离子模式离子化,多反应监测模式对美西律进行分析。结果美西律与内标纳洛酮分离良好,在0.02～10.00μg/mL内线性关系良好,相关系数为0.9999,回归方程为y=0.0283x-0.0151,日内与日间精密度的RSD均小于15%,最低检测限为0.01μg/mL。结论建立的LC-MS/MS方法简单、灵敏、可靠,可同时适用于美西律临床药物监测和法医毒物分析的需要。     

生物检材中吗啡类生物碱的LC-MS／MS分析

目的针对滥用药物分析鉴定实践中亟待解决的问题,开展LC-MS/MS分析生物检材中吗啡类生物碱的应用研究。方法满足不同的鉴定需要,分别建立血液、尿液、唾液和头发等生物检材的样品前处理方法,确定同时分析海洛因、单乙酰吗啡、吗啡、可待因、乙酰可待因、二氢可待因酮和氢吗啡酮等吗啡类生物碱的LC-MS/MS方法。将方法应用于实际案例。结果所建立的方法对吗啡类生物碱分离良好。尿液稀释法、尿液提取法和头发中吗啡的最低检测限(LOD)分别为10ng/mL、0.01ng/mL和0.01ng/mg。结论所建立的方法简便、快速、特异性强、灵敏度高。目标物中加入二氢可待因酮和氢吗啡酮扩大了方法的实用范围。     

Quantitative analysis of multiple illicit drugs in preserved oral fluid by solid-phase extraction and liquid chromatography-tandem mass spectrometry

We present a validated method for the simultaneous analysis of basic drugs which comprises a sample clean-up step, using mixed-mode solid-phase extraction (SPE), followed by LC-MS/MS analysis. Deuterated analogues for all of the analytes of interest were used for quantitation. The applied HPLC gradient ensured the elution of all the drugs examined within 14 min and produced chromatographic peaks of acceptable symmetry. Selectivity of the method was achieved by a combination of retention time, and two precursor-product ion transitions for the non-deuterated analogues. Oral fluid was collected with the Intercept, a FDA approved sampling device that is used on a large scale in the US for workplace drug testing. However, this collection system contains some ingredients (stabilizers and preservatives) that can cause substantial interferences, e.g. ion suppression or enhancement during LC-MS/MS analysis, in the absence of suitable sample pre-treatment. The use of the SPE was demonstrated to be highly effective and led to significant decreases in the interferences. Extraction was found to be both reproducible and efficient with recoveries >76% for all of the analytes. Furthermore, the processed samples were demonstrated to be stable for 48 h, except for cocaine and benzoylecgonine, where a slight negative trend was observed, but did not compromise the quantitation. In all cases the method was linear over the range investigated (2-200 microg/L) with an excellent intra-assay and inter-assay precision (coefficients of variation <10% in most cases) for QC samples spiked at a concentration of 4, 12 and 100 microg/L. Limits of quantitation were estimated to be at 2 microg/L with limits of detection ranging from 0.2 to 0.5 microg/L, which meets the requirements of SAMHSA for oral fluid testing in the workplace. The method was subsequently applied to the analysis of Intercept samples collected at the roadside by the police, and to determine MDMA and MDA levels in oral fluid samples from a controlled study.     

LC-MS／MS法测定人血浆中盐酸洛哌丁胺

目的建立人血浆中盐酸洛哌丁胺的液相色谱-质谱联用测定法（LC-MS／MS）。方法血浆样品中盐酸洛哌丁胺与盐酸小檗碱（内标）经甲醇液．液提取后,采用ZORBAXSB—C18色谱柱（2．1mm×150mm×5μm）,柱温35℃,流动相为乙腈：0．1％甲酸（60：40,V／V）,流速为0．4mL／min,进样量10μL。电喷雾离子源（ESI）,正离子检测,以多反应监测（MRM）方式进行定量分析,用于监测的离子为m／z477→266（盐酸洛哌丁胺）和m／z366→292（内标）。结果盐酸洛哌丁胺的检测下限为0．2ng／mL（S／N=3）,在浓度0．5～500ng／mL范围内线性良好（r=0．9982）,低、中、高浓度（1ng／mL、20ng／mL、400ng／mL）的平均回收率分别为84．6％,88．5％和90．2％,日内与日问RSD分别小于6％与7％。结论LC—MS／MS法可用于盐酸洛哌丁胺的定性定量分析。     

LC-MS/MS-MRM筛选血液中22种有毒生物碱成分

目的建立液相色谱-串联质谱法对血液中的22种常见有毒生物碱成分进行筛选分析。方法血液以丁丙诺菲为内标经液液提取后,用液相色谱-串联质谱仪以电喷雾电离(ESI )、多反应监测(MRM)方式进行分析。结果以化合物的保留时间、两对母离子/子离子对定性,最低检出限为0.1～20ng/mL。结论该方法选择性佳、灵敏度高,适用于法医毒物分析和临床毒物分析中有毒生物碱的分析。     

血中安定及其代谢物的酶水解研究

目的考察木瓜蛋白酶水解血中安定及其代谢物蛋白结合物的酶解条件,提高安定的提取率。方法采用正交试验确定酶解的最优条件,检材经蛋白酶水解,固相萃取后,应用LC-MS／MS方法进行检测,运用保留时间和MRM（多离子反应监测）方式来对血中安定及其代谢物进行定性定量分析。结果安定、去甲安定、去甲异安定、羟基安定和去甲羟基安定的最佳酶解条件分别为55℃,2．5h,pH7．0,8000U;50℃,1h,pH7．5,8000U;50℃,1h,pH7．5,8000U;50℃,1．5h,pH7．5,8000U;50℃,1．5h,pH7．5,8000U。结论酶水解后的血液中安定及其代谢物检出量明显增加。     

LC-MS/MS同时分析尿液中15种安眠镇静药物及其代谢物

目的建立尿液中15种常见安眠镇静药物及代谢物的液相色谱-串联质谱分析方法。方法尿液经酶水解、固相萃取后,用C18液相柱分离,以含甲酸铵和甲酸的水、乙腈为流动相梯度洗脱,质谱采用电喷雾电离（ESI）-正负离子模式同时扫描,采用二级质谱多反应监测（MRM）模式检测目标化合物。结果以化合物的保留时间、两对母离子/子离子对定性,尿中常见安眠镇静药物的检测限为0.01～0.5ng/mL（ESI＋）和10ng/mL（ESI-）;相关系数r在0.994以上;日内及日间精密度均在18%以下;绝对回收率在64.80%～116.20%之间。结论方法快速、灵敏、简便、可靠,能同时分析尿液中的15种安眠镇静药物及其代谢物。     

纵火现场中汽油、煤油和柴油残留物的ATD／GC／MS法检测

目的为火场中汽油、煤油和柴油残留物的检测建立一种简便、高灵敏度的ATD／GC／MS检验方法。方法采用Tenax TA吸附管吸附富集检材中的汽油、煤油和柴油成分。然后用美国PE公司的ATD／GC／MS仪器进行全自动的解吸和分析检测。结果检材中汽油、煤油和柴油的检测极限分别达0．05、0.2、0．2pL／mL。结论该方法具解吸和分析检测过程自动化。操作简便快捷，检测灵敏度高。杂质干扰少等特点，可用于实际火场皆汽油、煤油和柴油残留物的检测。     

吗啡及其代谢物在藏毒致死者体内分布初探

目的采用固相萃取、液相色谱一串联质谱（LC-MS／MS）检验方法,考察吗啡和葡萄糖醛酸吗啡（M3G）在一例体内藏毒致急性死亡者体内分布情况。方法提取死者心血、尿、胃内容物、肝、肾、脑等15种检材,经Waters HLB小柱固相萃取后,C18色谱柱分离,采用电喷雾电离（ESI）、多反应监测模式（MRM）检测目标化合物。结果所建方法在0．0l～101μg／mL浓度范围内线性关系良好,提取回收率大于75％。结果显示总吗啡含量（游离态＋结合态）在胃内容物中最高,其次是尿、‘肾,在心血、胃组织、肺和腺体中居中,脑组织和心脏含量最低。结论本例检验结果验证了胃内容物、尿液和肾脏等是该类中毒案件的理想检材,其分布规律也可作为体内毒品分析实验依据。     

固相萃取-液相色谱-质谱法检验人全血中的地塞米松

目的采用固相萃取结合液相色谱-质谱法检验人全血中地塞米松。方法采用SPE提取全血,用UPLC-MS/MS方法测定,采用ESI离子源,MRM方式监测。结果地塞米松的检出限为0.05ng/mL,线性范围1ng/mL~100ng/mL,方法回收率大于78.1%,日内、日间精密度均小于15%。结论 SPE-UPLC/MS/MS检验人全血中地塞米松的方法简单、高效,可应用于人全血中地塞米松的检验。     

LC-MS/MS法快速测定全血中25种精神药物的研究

目的建立人全血中25种精神药物快速测定的LC-MS/MS方法,并应用于分析杭州地区药物影响下驾驶(driving under the inference of drugs,DUID)情况。方法以乙腈沉淀蛋白,离心后取上清液,氮气流下吹干,残渣以初始流动相复溶,离心后取上清分析;采用C18色谱柱(50mm×3.0mm,2.6μm)分离,流动相:0.1%甲酸水(A相),乙腈:甲醇=1:1(B相),梯度洗脱;质谱检测,采用串联质谱电喷雾离子源,正电离扫描,多反应监测(MRM)。结果 25种精神药物在0.05~20ng/m L范围内线性良好,R=0.994 4~0.999 6;定量下限为0.05ng/m L;提取回收率为83.0%~99.7%;方法回收率为80.2%~97.4%;日内精密度(RSD)为1.6%~14%;日间精密度(RSD)为3.1%~14%。以该法测定杭州市公安司法鉴定中心留存的全血样品3140例,25种精神药物至少一种的检出率为3.7%。结论本方法灵敏、快捷、准确,适用于全血中25种精神药物快速检测。     

Associating gunpowder and residues from commercial ammunition using compositional analysis

Qualitatively identifying and quantitatively determining the additives in smokeless gunpowder to calculate a numerical propellant to stabilizer (P/S) ratio is a new approach to associate handgun-fired organic gunshot residues (OGSR) with unfired powder. In past work, the P/S values of handgun OGSR and cartridges loaded with known gunpowders were evaluated. In this study, gunpowder and residue samples were obtained from seven boxes of commercial 38 caliber ammunition with the goals of associating cartridges within a box and matching residues to unfired powders, based on the P/S value and the qualitative identity of the additives. Gunpowder samples from four of the seven boxes of ammunition could be easily differentiated. When visual comparisons of the cartridge powders were considered in addition to composition, powder samples from all seven boxes of ammunition could be reliably differentiated. Handgun OGSR was also collected and evaluated in bulk as well as for individual particles. In some cases, residues could be reliably differentiated based on P/S and additive identity. It was instructive to evaluate the composition of individual unfired gunpowder and OGSR particles. We determined that both the numerical centroid and dispersity of the P/S measurements provide information for associations and exclusions. Associating measurements from residue particles with those of residue samples collected from a test firing of the same weapon and ammunition appears to be a useful approach to account for any changes in composition that occur during the firing process.     

Elemental Characterization and Discrimination of Nontoxic Ammunition Using Scanning Electron Microscopy with Energy Dispersive X‐Ray Analysis and Principal Components Analysis

Concerns over the toxic by‐products produced by traditional ammunition have led to an increase in popularity of nontoxic ammunition. In this work, the chemical composition of six brands of nontoxic ammunition was investigated and compared to that of a road flare, which served as an environmental source with similar composition. Five rounds of each brand were fired while a further five were disassembled and the primer alone was fired. Particles collected from all samples, including the road flare, were analyzed by scanning electron microscopy with energy dispersive X‐ray analysis. Common elements among the different ammunition brands included aluminum, potassium, silicon, calcium, and strontium. Spectra were then subjected to principal components analysis in which association of the primer to the intact ammunition sample was generally possible, with distinction among brands and from the road flare sample. Further, PCA loadings plots indicated the elements responsible for the association and discrimination observed.