Screening and determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry

Benzodiazepines are one of the most widely prescribed drugs for the treatment of a wide spectrum of clinical disorders. They are used as anticonvulsants, anxiolytics, hypnotics or muscle relaxants with different duration of action. In this paper, a simple and sensitive method for the determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry (GC/MS) is described. The drugs spiked in whole blood were extracted with an Oasis HLB solid-phase extraction cartridge (Waters), which contains a copolymer designed to have a hydrophilic-lipophilic balance. GC/MS analysis was performed using a Shimadzu QP-5000 equipped with a BPX5 capillary column (15 mx0.32 mm I.D., film thickness 0.25 microm, SGE). Nineteen benzodiazepines and two thienodiazepines were well separated from each other on their SIM chromatograms and also on the TIC with the exception of oxazolam to cloxazolam separation. The blank extract from whole blood gave no peaks that interfered with all benzodiazepines and thienodiazepines on the chromatogram. The calibration curves for selected benzodiazepines with fludiazepam as an internal standard showed excellent linearity over the concentration range 5-500 ng/ml blood with a correlation coefficients of >0.995. The detection limits ranged from 0.2 to 20 ng/ml blood. The method is simple and sensitive for the determination of benzodiazepines in whole blood and seems to be useful in the practice of forensic science.     

A sandwich enzyme immunoassay for human pancreatic elastase III

To develop a method for the determination of pancreas injuries using a pancreas-specific antigen as a marker, human elastase III was purified from the pancreas by chromatographic methods. A rabbit anti-human elastase III antibody was prepared, and this antibody was confirmed using immunoblotting to react only with elastase III among proteins from the pancreas. A sensitive sandwich enzyme immunoassay for human elastase III was developed. The detection limit for human elastase III was 0.3 pg (10 amol) per assay. Proteins extracted from the pancreas showed the strongest response, whereas reactions of the other organs were less than the detection limit. These results suggest that a sandwich enzyme immunoassay for human elastase III is useful for the determination of pancreas injury.     

体外培养FBs损伤模型的建立及细胞免疫组化研究

首次应用培养人胎肺成纤维细胞 (FBs)损伤模型 ,体外研究创缘FBs合成细胞型纤维连接蛋白 (cellu larfibronectins,cFn)的变化与损伤时间的关系。结果表明 ,损伤使处于静息状态的融合FBs成为具有运动能力和增殖能力的活跃细胞。应用免疫组化ABC法结合图像分析技术 ,观察伤后不同时间创缘FBs内cFn的含量变化。伤后 1h ,可以检测到cFn有变化 ,且在伤后 6h内 ,cFn逐渐增多 ,其变化与损伤时间呈正相关。这对今后应用cFn与损伤时间推断的研究提供了科学依据 ,同时为法医学损伤时间研究开辟了新的途径。     

大鼠肝细胞DNA含量与早期死亡时间关系的图像分析研究

实验选择 15只大鼠 ,处死后 ,在 2 4h内 ,每隔 1h取肝组织块进行细胞学涂片、福尔马林液固定、Feul gen染色、自动图像分析仪测量、统计学处理数据。结果表明 ,肝细胞DNA含量在死后 2 4h内 ,随死亡时间的延长而较有规律的下降。其中积分光密度、平均光密度、异形指数是用来研究死亡 2 4h内肝细胞DNA降解规律 ,并准确推断死亡时间的较好指标。     

应用40Hz听觉相关电位与听性脑干反应评估听阈

对 42例 74耳（正常听力组 32耳,检案中主诉听力下降 42耳）分别进行纯音测定,声导抗测定,听性脑干反应 (Auditory Brainstem Response,ABR)及 0.5～ 2kHz范围的 40Hz听觉相关电位 (40Hertz Auditory Event－ related Potential,40Hz AERP)测定,对其中 20耳进行睡眠及清醒两种状态的 40Hz AERP测定。将纯音听阈、 40Hz AERP反应阈、 ABR反应阈三者之间进行比较分析。结果表明,正常听力耳 40Hz AERP反应阈值较纯音测听阈值高,在不同频率的校正值 (差值 )不同, 0.5 kHz为 12.7± 6.4(dBnHL); 1kHz为 14.7± 6.3(dBnHL); 2 kHz为 15± 5.6(dBnHL)。 ABR阈值比行为听阈高,校正值为 8.9± 5.3(dBnHL)。睡眠状态时 40Hz AERP阈值较清醒状态阈值高,校正值为 9.7± 2.45(dBnHL)。检案中主诉听力下降者主、客观语音频率均值之间明显不相符 ,误差率为 61％。本研究表明单纯使用 ABR、纯音测听中任何一种方法估计语音频率听阈均有一定的误差。     

损伤对培养人胎肺FBs分泌Fn的影响及与损伤时间的关系

为了探讨损伤对FBs分泌Fn的影响及与损伤时间的关系 ,在本室建立的体外培养人FBs损伤模型的基础上 ,应用酶联免疫组织化学检测技术 (双抗体夹心法ELISA)对损伤后不同时间培养液中Fn的含量变化进行检测 ,结果表明 :伤后培养人胎肺FBs合成分泌的cFn的量 ,在伤后6h内与损伤时间呈正相关。     

涉及死亡非医疗过失纠纷的类型,死因和发生原因——27年医疗纠纷法医尸检回?

对1992～1998年间206例非医疗过失尸检资料进行回顾性研究,表明涉及死亡的非医疗纠纷中各类疾病死亡占53.88%,难以避免的并发症占39.81%,医疗意外占6.31%.发生原因中对死因怀疑占44.66%,怀疑输液(血)错误占15.05%,怀疑治疗出问题占9.71%,怀疑用药错误或诊治延误各占8.74%,其它13.11%与医护人员态度不好、医院管理有关.     

中毒致死者体内芬氟拉明的GC/NPD,GC/MS法测定

建立了生物检材中芬氟拉明的定性定量分析方法。体液及脏器组织经有机溶剂提取后 ,用GC/MS法进行药物筛选、定性 ,生物检材中的芬氟拉明浓度用4 -苯丁胺作内标、GC/NPD法测定。测得芬氟拉明中毒致死者的血液、尿液、肝等组织中浓度分别为7.8μg/ml、64.2μg/ml、31.3μg/g。并对尸体解剖所见及方法可行性进行讨论