乌头生物碱及其代谢物在家兔死后体内分布研究

目的研究乌头生物碱及6种代谢物在中毒家兔死后体内的分布规律,为乌头生物碱中毒案件中检材的选取提供参考依据。方法将生川乌粉碎制成水煎剂,以其中乌头碱的LD50计算,给予家兔灌胃,待其死亡后立即解剖取材,采用高效液相色谱-串联质谱测定检材中乌头生物碱及6种代谢物的含量。结果乌头生物碱及6种代谢物在各脏器及体液中的死后分布情况为:(1)乌头碱:尿液外周血、心血、胃、肺、肾、心、肝、肌、脾;(2)新乌头碱:尿液外周血、心血、肺、胃、肾、肝、肌、脾、心;(3)次乌头碱:尿液外周血、心血、胃、肺、肝、脾、肾、肌、心;(4)苯甲酰乌头原碱:尿液外周血心血胃、脾、肾、肺、肌、心、肝;(5)苯甲酰新乌头原碱:尿液外周血心血胃、脾、肾、肺、肌、肝、心;(6)苯甲酰次乌头原碱:尿液外周血心血、胃、肾、脾、肺、肌、肝、心;(7)乌头原碱:尿液外周血、心血、肾、胃、心、脾、肝、肌、肺;(8)新乌头原碱:尿液外周血、心血、肾、胃、肝、心、肺、脾、肌;(9)次乌头原碱:尿液外周血、心血、肝、肾、脾、胃、心、肺、肌。结论乌头生物碱及6种代谢物分布以血液和尿液中为主,在脏器以胃、肺脏、肾脏和肝脏等含量较高。     

秋水仙碱在急性中毒豚鼠体液和组织中的检测及分布

目的建立生物检材中秋水仙碱的液相色谱/串联质谱检测方法,研究秋水仙碱急性中毒在豚鼠体内的分布,为秋水仙碱中毒的法医学鉴定提供方法和评价依据。方法以4mg/kg剂量秋水仙碱给豚鼠灌胃染毒,2～8h死亡,用LC-MS/MS测定体液和组织中秋水仙碱的含量。结果秋水仙碱在豚鼠各体液和组织中含量从高到低依次为胆汁、尿液、胃、脾、肺、肾、心、胰、肝、肾上腺、睾丸,心血中秋水仙碱的含量最低。结论所建方法准确、灵敏,适用于体内痕量秋水仙碱的检测。胆汁和尿液是体内检测秋水仙碱的较佳检材。     

LC-MS~n分析新乌头碱在兔尿液中的代谢产物

乌头属(aconitum)中药临床应用广泛,治疗风湿痹痛等症疗效显著。但该属中药所含乌头类生物碱既是有效成分又是剧毒成分,其治疗量与中毒、致死量接近[1],因此服用过量、误服或投毒等都可引起乌头类生物碱中毒[2,3]。乌头碱中毒无特异性临床表现,且在体内迅速分解,给检测带来困难。在乌头属中药中新乌头碱(mesaconitine)多与乌头碱(aconi-tine)共存,且新乌头碱含量高于乌头碱。本文应用LC-MSn分析家兔灌服新乌头碱中毒死亡前尿液中的代谢产物,为检测新乌头碱的代谢产物提供新方法。1材料与方法1.1仪器与材料Finnigan公司LCQ液相色谱-质谱…     

乌头属植物生物碱高效液相色谱研究

目的建立高效液相色谱法测定多种乌头属植物生物碱的方法。方法XTerraTMRP185μm,(250mm×4.6mm)色谱柱;流动相为乙腈∶30mmol/L碳酸氢铵梯度洗脱;流速1.0ml/min;检测波长233nm。结果该方法可成功应用于5种乌头属植物12个样品中生物碱成分分析。结论本研究结果提示乌头属不同种植物中生物碱成分及含量差异显著,因此为确保该类中药材用药安全,提高临床疗效,必须建立严格的药材质量控制体系。     

乌头中剧毒生物碱的提取分离

目的从乌头属植物中提取分离剧毒生物碱乌头碱、中乌头碱、次乌头碱;方法酸性水溶液提取,碱化后中性氧化铝柱层析,IR、EI-MS确认结构;结果利用该方法提取以上3种生物碱得到的回收率与碱性苯溶液法相似。结论该方法所用试剂成本低、毒性小,适用于制备乌头碱、中乌头碱、次乌头碱对照品。     

乌头属植物生物碱高效液相色谱研究Ⅱ

目的 建立高效液相色谱法测定多种乌头属植物生物碱的方法 .方法 XTerraTMRP185μm,(250×4.6mm)色谱柱;流动相为乙腈:30mmol/L碳酸氢铵梯度洗脱;流速1.0ml/min;检测波长233nm.结果 该方法可成功应用于黄草乌、紫草乌、短距牛扁、保山乌头4种乌头属植物生物碱成分分析.结论 本研究结果提示乌头属植物中毒案件检验应注意是否含有8-O-乙酰基-14-茴香酰基生物碱.     

药酒中乌头碱、次乌头碱、新乌头碱的LC/MS/MS分析

目的应用高效液相色谱-质谱联用法对涉案药酒中有毒成分乌头碱、次乌头碱、新乌头碱快速定量分析。方法样品过膜后用乙腈稀释500倍,使用Waters ACQUITY UPLC BEN C18(100 mm×2.1 mm,1.7μm)色谱柱,以乙腈-10 mmol/L碳酸氢铵溶液(pH=10)为流动相梯度洗脱,多反应监测模式下测定涉案药酒中乌头生物碱的含量。设定方法下乌头碱、次乌头碱、新乌头碱的保留时间分别为5.10 min、5.64 min、4.58 min,用于定量分析的离子对(m/z)分别为646.6-586.6、616.6-556.6、632.7-572.6。在1~200 ng/mL内,乌头碱线性回归方程为Y=343.8X+38901(R2=0.9995);在100~1000 ng/mL内,次乌头碱线性回归方程为Y=3.817X-357.579(R2=0.9991);在0.5~10 ng/mL内,新乌头碱线性回归方程为Y=719.562X+128.748(R~2=0.9995)。用此方法检测出涉案药酒中乌头碱、次乌头碱、新乌头碱含量分别为71.85,415.86,1.49μg/mL。结论建立的高效液相色谱-质谱法可用于药酒中乌头碱、次乌头碱、新乌头碱的快速测定。     

氯胺酮在家兔死后体内的弥散研究

目的研究氯胺酮在大白兔体内死后弥散过程和再分布机制。方法 48只实验大白兔随机分为8组,采用缺氧处死后以150mg/kg氯胺酮灌胃,尸体仰卧位于室温下放置;在0~96h内分8个时间点各解剖1组,提取体液和脏器组织样品;采用GC/MS法定性结合GC-NPD法定量检测样品中氯胺酮含量,并计算心血/外周血中氯胺酮含量的比值。结果大白兔死后氯胺酮灌胃尸体放置96h内,脑、尿液、玻璃体液、左上/下肢肌肉样本中均未检测到氯胺酮,心血、外周血、心肌、脾、肾、肝、肺、胆汁中氯胺酮含量随死后时间呈动态升高的变化;其中距离胃较近的组织(如脾)较早检测到含量较高的氯胺酮,而距离较远的组织或体液中氯胺酮含量较低且较晚检测到;心血/外周血中氯胺酮含量比值为1.73。结论氯胺酮在家兔体内存在死后再分布,从胃到器官组织、心血顺浓度梯度弥散是主要机制。脑、玻璃体液、尿液、肢体肌肉不受死后弥散的影响,可作为生前服毒与死后染毒氯胺酮的鉴别依据。     

超高效液相色谱-质谱联用法测定生物检材中乌头碱

目的建立一种简单、快速测定生物检材(血液、尿液、胃内容物)乌头碱的超高效液相色谱-质谱联用法。方法样品采用乙腈沉淀处理。色谱柱为UPLC C18(2.1mm×50mm,1.7μm),流动相为乙腈-0.1%甲酸,梯度洗脱,流速为0.4m L/min,柱温40℃。采用ESI离子源,MRM模式检测。结果乌头碱在血液0.5~500ng/m L,尿液1~1000ng/m L浓度范围内线性良好(r>0.995)。乌头碱方法回收率在91.3%~110.2%之间;提取回收率在72.8%~83.5%之间;日内、日间RSD均小于14%。结论本方法简单、快速检测生物检材中乌头碱。     

短柄乌头急性中毒——乌头碱在大鼠体内分布的研究

本文用高效液相色谱法检测了大白鼠短柄乌头急性中毒后(LD_(50)剂量灌胃),乌头碱在心、肝、肾、血、脑内的含量分布。结果表明;体内乌头碱含量甚微或检不出。30例大鼠LD_(50)剂量灌胃后,16例2h内中毒死亡。其肝、肾内的乌头碱检出率明显高于心、血、脑,且中毒表现明显。另14例于2h整处死,其肝、肾、血中检出较高。16例2h内死亡组肝中乌头碱含量分析提示乌头碱在体内代谢很快。此结果为法医工作中乌头碱中毒案件调查、检材提取、毒物分析结果的评价提供了一定的依据。     

Aconitum alkaloid content and the high toxicity of aconite tincture

Although proprietary medicines and decoction of processed aconite roots are the most widely used, tincture accounts for the great majority of aconite poisoning cases in China, indicating that it is much more toxic than other formulations. Aconite tincture is often self-prepared at home and raw aconite plants or roots are often used. Even if processed aconite roots were used to make the tincture, the amount of Aconitum alkaloids is highly variable, depending on the adequacy of processing and quality control. Aconitum alkaloids dissolve efficiently in alcohol. For these reasons, tincture contains very high concentrations of Aconitum alkaloids. Despite its high intrinsic toxicity, overdose of aconite tincture by the users has been common. Severe aconite poisoning can be complicated by fatal ventricular tachyarrhythmias and asystole. The public should be repeatedly warned of the danger of taking aconite tincture by mouth.     

体液中微量士的宁的富集及同时检测

目的将分子印迹技术运用到法医毒物分析中,建立一种全新的测定体液中微量士的宁的方法。方法运用分子印迹原位聚合技术,在色谱柱中一步合成了对士的宁具有专属识别能力的分子印迹聚合物,以此聚合物为介质,装填一厘米的小柱,建立体液中微量士的宁的富集及同时检测方法。结果以士的宁分子印迹聚合物作为尿液和血浆中微量士的宁的富集和检测介质,方法的检出限为4.9ng,样品回收率均大于92%,相对标准偏差小于6.59%,工作曲线的线性相关系数分别为0.9991和0.9966。运用此方法对中毒家兔血浆和尿液中的士的宁进行了测定。结论建立的新方法可以实现对士的宁专属的、灵敏的检测,适合于法医学毒物分析运用。     

Seven cases of fatal aconite poisoning: forensic experience in China

This paper presents seven fatal cases of aconite poisoning encountered in the Tongji Center for Medicolegal Expertise in Hubei (TCMEH), China, from 1999 to 2008 retrospectively. In six of the cases, deaths occurred after drinking homemade medicated liquor containing aconite, and in one case death was due to ingestion of traditional Chinese medication containing aconite. Forensic autopsy and pathological examinations ruled out the presence of physical trauma or life-threatening diseases. Diagnosis of aconite poisoning was made after postmortem toxicological analysis. Animal experiment was performed in one case demonstrating that the medicated liquor could cause death rapidly. We present the autopsy and histopathological findings, toxicological analysis, and results of animal experiment done on samples from those seven cases. As an important herbal Chinese medicine, Aconitum species deserve special attention, especially because it contains poisonous alkaloids.     

γ-羟基丁酸在急性中毒大鼠体液和组织中的检测及分布

目的建立生物检材中γ-羟基丁酸(GHB)的检测方法,研究GHB急性中毒大鼠体内GHB的分布,为GHB中毒的鉴定提供方法和评价依据。方法用GC/MS法检测生物检材中的GHB;以1000mg/kg剂量给大鼠灌胃使其染毒,分别于1h和3h处死,测定体液和组织中GHB的含量。结果测组织中内源性GHB的线性范围是1～20μg/g,R2=0.9974;测组织中外源性GHB的线性范围为100～1500μg/g,R2=0.9958。相对回收率为98%～103%。体内内源性GHB的含量均≤10μg/mL或10μg/g。尿液中GHB含量为最高,其他依次为:胃、血液、肠、肾、肺、脾、心、肝和脑。结论所建方法准确、便捷,适用于GHB中毒的鉴定;尿液是体内检测GHB的最佳检材。     

Lithium intoxication. A case study

This study reports a fatal ingestion of lithium carbonate and its distribution in tissues and biological fluids.     

A fatal case of oral ingestion of toluene

A 51-year-old male ingested orally a large quantity of toluene and died about 30 min later. The presence of toluene in body fluids and tissues was confirmed by gas chromatography and gas chromatography/mass spectrometry. Tissue distribution of toluene showed that the liver detected the highest content of toluene (433.5 micrograms/g), except for the stomach contents, followed by pancreas (88.2 micrograms/g), brain (85.3 micrograms/g), heart (62.6 micrograms/g), blood (27.6 micrograms/g), fat (12.2 micrograms/g) and finally cerebrospinal fluid (11.1 micrograms/g).     

A fatal case of oral ingestion of methanol. Distribution in postmortem tissues and fluids including pericardial fluid and vitreous humor.

A 41-year-old man ingested orally a large quantity of methanol and was found dead at home. The presence of methanol in body fluids and tissues was determined by head-space gas chromatography. The blood ethanol and acetone were negative. Tissue distribution of methanol showed that the kidney presented the highest content of methanol (5.13 g/kg) followed by liver (4.18 g/kg), vitreous humor (3.96 g/l), heart (3.45 g/kg), urine (3.43 g/l), pericardial fluid (3.29 g/l), blood (2.84 g/l) and finally stomach content (2.21 g/l).     

Lethal poisoning by zipeprol in drug addicts

Two cases of lethal intoxication involving or due to oral ingestion of zipeprol are described. The two cases concerned abusers of the substance for nonmedical purposes. Data regarding the distribution of the unmodified drug in biological fluids and tissues are presented.     

Quantitative analysis of tropane alkaloids in biological materials by gas chromatography-mass spectrometry

A simple and rapid method for quantitation of tropane alkaloids in biological materials has been developed using an Extrelut column with gas chromatography-mass spectrometry (GC-MS). Biological materials (serum and urine) were mixed with a borate buffer and then applied to an Extrelut column. The adsorbed tropane alkaloids were eluted with dichloromethane before a GC-MS analysis. Atropine-d(3) was used as an internal standard. The extracted tropane alkaloids were converted to trimethylsilyl derivatives prior to GC analysis, to improve the instability of tropane alkaloids from heating and the property of them for a GC column. The recoveries of the compounds, which had been spiked to biological materials, were more than 80%. The GC separation of the derivatives from endogenous impurities was generally satisfactory with the use of a semi-polar capillary column. Tropane alkaloids showed excellent linearity in the range of 10-5000 ng/ml and the limit of detection was 5.0 ng/ml for biological materials. The present method is simple and more rapid than those previously reported, and was applied to a poisoning case. It is useful for the routine analysis of tropane alkaloids in cases of suspected tropane alkaloids poisoning.