Detection and quantification of psychotropic drug etaqualone in human hair using GC–MS/MS

In this study, sensitive analytical procedure for detection and quantification of etaqualone in human hair samples using gas chromatography tandem mass spectrometry (GC–MS/MS) was newly established, and applied it to authentic human samples obtained from an abuser. In this method, the hair samples were treated with hydrochloric acid and then extracted with ethyl ether. The ether layer was dried in a warm water bath, and the residue was reconstituted in ethyl acetate, followed by GC–MS/MS analysis. Multiple reaction monitoring (MRM) mode was used for data collection, and quantitative analysis was performed using internal standard method. Good linear relationship within the concentration range of 1–100 pg/mg were obtained in calibrators for the hair samples showing its correlation coefficient value was 0.9993. The lower limit of quantitation in this study was 1 pg/mg and the recovery rate examined ranged from 100.4% to 108.5%. The intra-day precision and accuracy were less than 5.0% and 5.8%, respectively. The inter-day precision and accuracy were lower than 6.4% and 4.6%, respectively. Using this established method, etaqualone could be detected in the hair sample obtained from a suspected user to be level of 65.2 pg/mg. It should be expected that the method established in this study would contribute to rapid detection and identification of psychotropic drug etaqualone among multiple fields including forensic investigation, clinical application and of course public health matters.     

Determination of dihydrocodeine in hair of opiate addicts by GC/MS

Summary After the examination of more than 300 hair samples of suspected heroin abusers, a large number of which proved positive, we can say that high concentrations of dihydrocodeine can be determined either in addition to, or in the place of, morphine and also frequently in combination with codeine. The opiates were extracted after dissolving the hair samples in NaOH and hydrolysis with HCI. The quantitative determination of dihydrocodeine was achieved by derivatisation with HFBA using GC/MS at m/u = 497. Dihydrocodeine is used in antitussive drugs. After the examination of individual hair samples, it was obvious that some heroin consumers had switched to dihydrocodeine. This may lead to the conclusion that dihydrocodeine itself is used either as an intoxicating drug or to reduce withdrawal symptoms. The increasing number of positive samples should be noted by the legal authorities.     

Development of a preparation method to produce a single sample that can be applied to both LC–MS/MS and GC–MS for the screening of postmortem specimens

Simple and efficient extraction methods have been developed for the screening of a wide array of drugs in postmortem autopsy specimens. Acidic and basic compounds were targeted with two extraction methods that can be applied to both GC–MS and LC–MS/MS instrumentation. The extractions were achieved by utilizing lipid-removal and solid-phase extraction cartridges while carefully monitoring the pH of the samples to ensure the adequate removal of interfering substances like lipids and amino acid derivatives. These methods were applied to actual autopsy cases, with 94 and 124 compounds detected by GC–MS and LC–MS/MS, respectively. The developed methods could easily be incorporated into a forensic laboratory’s daily routine for screening many different compounds from postmortem samples.     

LC/MS/MS法测定法罗培南血尿药浓度及药动学研究

目的研究法罗培南钠片在健康人体内的药动学及饮食对药动学的影响。方法药动学试验：采用低、中、高三个剂量组的平行试验设计；饮食对人体药动学的影响试验：采用自身对照的双周期交叉试验设计。血浆样品处理后采用LC／MS／MS测定血药浓度。结果空腹po100、200、400mg法罗培南片，主要药动学参数AUC和Cmax随剂量呈线性相关。进餐后影响药物的Tmax和Cmax，其他药动学参数无变化。给予法罗培南在100、200、400mg的剂量，受试者尿中0～24h原形药物的平均累积排泄率分别为4．52％、7．73％和3．28％。结论LC／MS／MS方法操作简单，结果准确，专属性强，灵敏度高，适用于法罗培南人体药动学研究。     

Interpretation of forensic cases based on empirical data

Over the course of the last years the importance of hair analysis increased obviously. For example work place testing, driving licence cases, so-called health checks, and especially in criminal acts the analysis of illicit substances is common. With the modern multiplex analytical methods (GC/MS, GC/MS/MS, LC/MS) the routine analytical probe of hair is in general unproblematic. But one of the major problems in hair analysis is the interpretation of the results. To solve this difficult and challenging task statistical data as a source of information could be a helpful tool. Examination of more than 1,000 hair segments within the past 2 years using gas chromatography/tandem-mass spectrometry (GC/MS/MS) as the preferred method for conducting the daily routine detection of the most prevalently abused drugs and metabolites, such as cannabinoids, cocaine, some synthetic drugs from the amphetamine group, and opiates serve as the basis for the statistics. The quantity of analytical results and additional essential facts have been documented in the publication to present our sources of information and supplementary data to prepare forensic reports. We hereby state that we do not have a significant financial interest or other relationship with any product manufacturer or provider of services discussed in this article.     

禁用药物β_2-激动剂的分析研究

采用灵敏度很高的气相色谱 /负离子化学电离质谱 (GC/NICIMS)对 9种国际奥委会禁用药物 β2 -激动剂进行分析研究 ,比较了不同衍生化方法得到的产物在NICIMS上的响应。确定了五氟丙酸酐衍生化产物有最佳的灵敏度 ,适合此类药物的检测要求。重点对克伦特罗尿样进行了检测 ,其结果符合实际工作的需要     

Determination of cocaine in human hair by gas chromatography/mass spectrometry

Summary A qualitative method for the determination of cocaine alone without its metabolites in human hair by gas chromatography/mass spectrometry (GC/MS) was developed. The assay used helium as carrier gas, a 30-m bonded phase fused silica OV-1 capillary column, and solid injection at 290°C evaporator temperature.The cocaine concentrations in hair were determined also by radio-immunoassay (RIA). The values obtained are the sum of cocaine and its metabolites.Both GC/MS and RIA meet the requirements for the determination of drug abuse by two different methods in forensic science.     

LC／MS／MS法测定中国健康志愿者血浆中瑞舒伐他汀浓度及药动学

目的 建立人血浆中瑞舒伐他汀的液相色谱-质谱-质谱联用测定方法，研究中国健康人体药代动力学。方法 以氢氯噻嗪为内标物，采用液相色谱-质谱-质谱联用法，电喷雾电离源选择性正离子峰检测。测30名健康男性志愿者单剂量口服瑞舒伐他汀钙片的体内血药浓度，获得药动学参数。结果 口服瑞舒伐他汀钙片5、10、20mg后的主要药代学参数：达峰时间Tmax分别为（3．10±0．99）、（2．00±0．82）、（3．40±1．58）h；峰值血浆浓度Cmax分别为（8．32±2．44）、（14．8±3、97）、（20．1±5．0）ng·ml^-1；药时曲线下面积AUC0-∞分别为（77．0±22．6）、（153±38）、（270±61）ng·h·ml^-1；AUCo．1分别为（75．8±22．0）、（151±37）、（258±62）ng·h·ml^-1；T1／2分别为（13．0±3．9）、（12．5±3．5）、（18．5±4．9）h。结论 受试者口服瑞舒伐他汀钙片后，在人体内表现为线性药代学特征。     

LC/MS/MS法测定家兔血浆和房水中咪哒酚衍生物含量及其药代动力学研究

目的建立准确、灵敏、可靠的LC/MS/MS方法,定量检测家兔血浆和房水中咪哒酚衍生物浓度,并用于家兔给予咪哒酚衍生物滴眼后血浆及房水中药代动力学研究。方法血浆及房水样品经蛋白沉淀前处理,内标选用Amms,液相色谱柱为TOSOHTSKgel Amide,流动相为含0.1%甲酸的甲醇∶水（80∶20,体积比）,仪器选用Finni-gan公司TSQ Quantum型液相色谱-质谱联用仪,采用电喷雾离子化电离源（ESI）,选择反应离子监测（SRM）方式进行正离子检测,用于定量分析的离子反应分别为m/z 381→308（咪哒酚衍生物）和m/z 285→138（Amms）。结果在本实验条件下,咪哒酚衍生物与内标Amms分离良好,保留时间分别为1.48和1.55 min,血浆和房水中药物浓度分别在5~2000和10~10 000 ng/ml范围内线性关系良好,相关系数（r2）分别为0.9971和0.9999,定量下限分别为5和10 ng/ml,回收率均大于80%,日内、日间精密度均小于15%（n=5）。结论该方法快速、准确、灵敏度高,专属性好,可用于咪哒酚衍生物在家兔体内的药物代谢动力学研究。     

GC／MS法测定人血浆中地芬尼多药物浓度及其在药代动力学研究中的应用

目的建立GC/MS法测定人血浆中地芬尼多的含量,用于药代动力学研究中血药浓度的测定。方法液-液萃取法进行血样前处理,分析采用（15 m×0.25 mm×0.25μm）DB-17MS毛细管柱,载气为氦气,载气流速为1.0 ml/m in;进样口温度250℃,炉温升程：100℃→280℃（按25℃/m in的速度升温,且在280℃保持1 min）再升至300℃（按50℃/min的速度升温,且在300℃保持1.6 m in）;MS源温度230℃,四级杆温度150℃,电子轰击能量为70 eV。结果地芬尼多及内标的保留时间分别为7.8 min及7.97 min,其定量碎片基峰分别为m/z98和138。地芬尼多在1300 ng/ml浓度范围内,浓度与响应呈良好的线性关系（r2=0.999）;定量下限为1 ng/ml,准确度在82%104%范围内;地芬尼多的提取回收率为72.7%90.0%;批内和批间RSD分别在2.7%和3.9%以内,准确度在101.5%114.1%范围内;地芬尼多人血浆样品在-20℃保存10 d及在室温放置24 h稳定;其甲醇溶液在4℃冰箱保存21 d稳定。结论本研究建立的GC/MS方法测定人血浆中地芬尼多浓度,特异性好、灵敏度高,适用于盐酸地芬尼多口崩片在健康受试者体内的药代动力学研究。     

LC／MS／MS测定比格犬血浆中丁螺环酮浓度及其在药代动力学研究中的应用

目的：建立LC／MS／MS测定比格犬血浆中丁螺环酮的方法,并应用于其缓释制剂药代动力学研究。方法：血浆样品经液-液萃取,内标选用曲马多,色谱柱采用Zorbax SB-C18柱（2．1mm×100mm,3．5μm）;流动相为甲醇-水-甲酸（40：60：0．1）,流速0．2ml／min;用美国Finnigan公司TSQ Quantum型液相色谱-质谱联用仪器,配有电喷雾离子化电离源（ESI）,正离子检测模式,扫描方式为选择反应监测（SRM）,用于定量分析的离子反应分别为m／z 386．2→121．97（丁螺环酮）和m／z 264．3→58．0（内标,曲马多）。结果：测定丁螺环酮的线性范围为0．025→7．5μg/L,相关系数为0．99893,提取回收率为72．2％,方法回收率为99．1％,日内日间精密度均小于10．7％,血浆中丁螺环酮的最低定量下限为0．025μg／L。结论：该方法简便、快速、准确,灵敏度高,专一性好,成功应用于丁螺环酮缓释制刺的药代动力学研究。     

液相色谱-串联质谱法测定人血浆中拉米夫定浓度

 目的 试图建立测定人血浆中拉米夫定浓度的液相色谱-串联质谱法.方法 以吉西他滨为内标,内标法定量.流动相:甲醇-10 mmol/L乙酸铵(12: 88,v/v);质谱采用离子喷雾离子源,扫描方式为多反应监测(MRM),用于定量分析的离子反应分别为m/z 230m/z 111.9(拉米夫定)和m/z 263.8m/z 111.9(吉西他滨);药动学参数采用DAS2.0软件处理获得.结果 拉米夫定和内标吉西他滨的保留时间分别为2.9 min和2.4 min;拉米夫定的线性范围为0.0100~2.0000 μg/ml ,r=0.9996,回归方程:Y=0.0032X+0.0003,最低检测浓度为0.0100 μg/ml;提取回收率为95%～105%范围内;日内日间精密度RSD<6%.结论 此法适合人体血浆拉米夫定浓度的监测及生物利用度研究,结果准确、可靠.     

Specific interpretation of hair concentrations in 2 fatal metformin intoxication cases

Hair analysis is very useful for toxicological investigations since, by providing a wider detection window, it gives the possibility to perform a retrospective study on the historical consumption of a substance. Unfortunately, there are no data available for hair concentrations in metformin-related deaths. In this study, the authors present 2 cases of fatal metformin intoxication in which, for the first time, hair analysis was performed using a specific GC–MS/MS method. Metformin was tested positive in femoral blood (112.3 mg/L and 64.7 mg/L respectively) and cardiac blood (226.9 and 203.2 mg/L) of the two subjects. For case 1, other samples were also tested positive, including vitreous humor (31.1 mg/L) and gastric contents (773.5 mg/L). In case 2, metformin was measured at 844.9 mg/L in urine. Metformin hair concentrations were 28.3–44.8 and 22.5 ng/mg for both cases, respectively. The concentrations found in the 2 fatal cases are clearly higher than those obtained in a previous study with subjects under treatment (0.3–3.8 ng/mg) or those found in 3 post-mortem cases where metformin death was excluded (0.6–1.4 ng/mg). Excessive sweating during the agonal phase due to fatal hypoglycemia could explain these elevated concentrations as sweat can have contaminated the hair.     

Simultaneous quantification by LC/ESI–MS/MS of chlorinated tyrosine derivatives in the autopsy sample of a victim of chlorine exposure

Direct detection and accurate quantification of chlorine in autopsy samples are difficult because of the volatility and rapid metabolism of chlorine. Here, we developed and validated a method for quantitative analysis of 3-chloro-l-tyrosine (Cl-Tyr) and 3,5-dichloro-l-tyrosine (DiCl-Tyr) as stable markers of chlorine exposure. Chemical derivatization followed by liquid chromatography coupled with electrospray ionization–tandem mass spectrometry (LC/ESI–MS/MS) enabled us to simultaneously analyze both Cl-Tyr and DiCl-Tyr in an autopsy sample from the victim of chlorine exposure. Cl-Tyr was detected in the heart blood (53.6 ng/mL), urine (9.5 ng/mL), and lung tissue (211.1 ng/g); however, DiCl-Tyr was detected only in the lung tissue (10.3 ng/g). In contrast, in autopsy samples obtained from cases without exposure to chlorine, DiCl-Tyr was not detected in any matrixes. Our result suggested that the simultaneous detection of Cl-Tyr and DiCl-Ty may provide a better appreciation of chlorine exposure. To our knowledge, this is the first time Cl-Tyr and DiCl-Tyr have been determined simultaneously in a real human autopsy sample from a victim of chlorine exposure.     

猪肉及其可食性组织中残留克伦特罗的GC/MS检测方法

目的 :建立猪肉及其可食性器官中克伦特罗的检测方法。方法 :肌肉或组织匀浆后经有机溶剂提取 ,提取液经碳 1 8小柱除去杂质 ,洗脱液在氮气下吹干后用MSTFA衍生化 ,衍生化液直接供GC/MS检测。结论 :利用此方法可以检测猪肉及其可食性器官中的克伦特罗 ,其检测限为 1ng/g。     

Direct detection of serum psilocin glucuronide by LC/MS and LC/MS/MS: time-courses of total and free (unconjugated) psilocin concentrations in serum specimens of a “magic mushroom” user

Psilocin glucuronide (PCG) was directly identified in serum specimens of a “magic mushroom” user by liquid chromatography/mass spectrometry (LC/MS) and LC/MS/MS, together with the free (unconjugated) psilocin (PC). A major part of serum PC existed in the conjugated form. To quantify the total (conjugated plus free) PC in serum, enzymatic hydrolysis conditions were optimized using the user’s urine as the source of PCG; PCG in serum could be completely hydrolyzed by Escherichia coli β-glucuronidase. Using the established procedure, both total and free PC in the serum specimens of the user collected at various intervals were quantified. For the first specimen collected 5h after magic mushroom ingestion, 71.0 ng/ml of total PC and 13.3 ng/ml of free PC were detected. The ratio of free PC to total PC decreased with time after ingestion. The β-glucuronidase treatment of serum was found to clearly extend the detectable period of the serum PC; PC could be detected even 52 h after ingestion of magic mushroom.     

Testing of skeletonized human remains using GC/MS – development of a personal environmental profile

ABSTRACT

Gas chromatography/mass spectrometry (GC/MS) is well used in forensic science for the identification of materials present in toxicology, pharmacology and arson investigation. However, GC/MS has been underused in human identification. In this study, highly commingled skeletonized remains from a mass fatality incident involving over 400 individuals were examined. Environmental compounds and those present in the remains themselves were extracted from osseous materials removed from the surface of the skeletal elements using two different solvents, dichloromethane and acetonitrile. Solvent extracts were concentrated and resuspended in methanol prior to injection on the GC/MS instrument. Compounds present from the environment surrounding the individual post-mortem, as well as biological materials present in the individual ante-mortem, are detectable. Traces generated from the GC/MS instrument provide distinctive images, and analysis of those materials shows patterns that are specific to the individual and may be used for individuation. Results indicate that GC/MS analysis of skeletonized remains may be a new tool for human identification.     

HPLC-MS/MS法测定人血浆中左西孟旦

目的建立快速、灵敏的HPLC-MS/MS法测定人血浆中的左西孟旦浓度。方法以氢氯噻嗪为内标,血浆样品经甲基叔丁基醚沉淀蛋白,以甲醇为流动相,采用C18色谱柱进行分离,电喷雾离子化源,以多重反应监测方式进行负离子检测。结果测定血浆中左西孟旦方法的线性范围为0.5～1200 ng/ml,定量下限可达0.5 ng/ml。结论本方法专属性好,准确、快速,适用于左西孟旦注射液的临床药代动力学研究。     

液质联用法测定龙胆水煎剂中三种环烯醚萜的含量

目的利用液质联用（LC／MS／MS）方法测定龙胆水煎剂中环烯醚萜类化合物龙胆苦苷、獐牙菜苦苷和马钱苷的含量。方法色谱柱为J’Sphere ODS-H80（150mm×2．0mm，4μm）；流动相为甲醇-10mmol·L^-1醋酸铵溶液（30：70）；流速为200μl／min。样品经电喷雾离子源（ESI）正离子化后，通过三级四极杆串联质谱仪，采用多反应离子监测方式测定样品中龙胆苦苷（m／z 357．2-195．3）、獐牙菜苦苷（m／z 392．1→195．2）和马钱苷（m／z 408．4→229．1）浓度。结果在该分析条件下，3种化合物能够同时进行检测。龙胆苦苷浓度在200～5000ng·ml^-1范围内线性关系良好，回归方程为Y=1．26×10^3X＋2．27×10^3，r=0．9983；獐牙菜苦苷浓度在20-500ng·ml^-1范围内线性关系良好，回归方程为Y=360X＋144，r=0．9991；马钱苷浓度在20-500ng·ml^-1范围内线性关系良好，回归方程为Y=782X＋42．6，r=0．9994。对照品峰面积精密度RSD分别为龙胆苦苷2．52％，獐牙菜苦苷2．77％和马钱苷2．89％；平均加样回收率分别为龙胆苦苷98．2％，獐牙菜苦苷99．1％和马钱苷99．7％。结论该方法灵敏、准确、特异性强。     

A fatal clomipramine intoxication case of a chronic alcoholic patient: application of postmortem hair analysis method of clomipramine and ethyl glucuronide using LC/APCI/MS

Toxicological investigations of postmortem specimens of a 26-year-old man were performed with the use of LC/APCI/MS. They revealed in the blood of the deceased clomipramine (9.49 microg/g) and its main metabolite norclomipramine (1.10 microg/g) at concentrations explaining the fatal outcome. The presence of these xenobiotics in a 12-cm-long strand of hair (clomipramine, 7.60 ng/mg in I segment; 4.19 ng/mg in II segment; 1.86 ng/mg in III segment; norclomipramine, 5.71 ng/mg in I segment; 9.71 ng/mg in II segment; 4.13 ng/mg in III segment) confirmed the fact obtained from the medical history that the deceased had been receiving clomipramine as an antidepressant for 1 year prior to his death. The analysis demonstrated ethanol in autopsy blood (2.5mg/ml) and urine (3.2mg/ml); ethyl glucuronide as a marker of chronic alcohol abuse was detected in the deceased's hair (0.44 ng/mg in I segment; 0.07 ng/mg in II segment; n.d. in III segment). These findings may suggest the contribution of alcohol in the mechanism of drug-ethanol interaction, which in consequence might have affected the biotransformation of clomipramine in the final period of his life and evoked the ultimate toxic effect.