全血中度冷丁的气—联用法测定及代谢动力学的研究

应用气－质联用选择离子法测定全血中度冷丁的浓度。方法线性范围为０．００２～１ｎｇ／ｍｌ（ｒ＝０．９９９７），变异系数小于１０％，最低检测限过１ｎｇ／ｍｌ，无杂质干扰。本法已用于肌注８０ｍｇ度冷丁后代谢动力学的研究。     

大麻的体内代谢物THC—COOH的分析

本文介绍了定性定量分析大麻的主要代谢物ＴＨＣ－ＣＯＯＨ的两种方法。所有试管硅烷化，尿样碱性水解。ＴＬＣ法快速、简便，适用于多份检材的同时定性分析，最低检测限为５０ｎｇ．采用衍生化、ＳＩＭ的ＧＣ／ＭＳ方法灵敏，选择性、专一性强．五氟丙酸酐衍生化方法线性范围为２０－２５０ｎｇ／ｍｌ，最低检测限１０ｎｇ／ｍｌ，变异系数为８．４％，提取回收率为５８．９％；甲基化方法线性范围１０～１００ｎｇ／ｍｌ，最低检测限５ｎｇ／ｍｌ，变异系数为５．６％，提取回收率为５３．２％．     

甲胺磷及其代谢产物检验的研究

本文利用ＧＣ／ＭＳ和ＧＣ／ＦＰＤ法测定了生物体内的甲胺磷（ＭＡＰ）及其代谢物Ｏ、Ｓ－二甲基磷酸酯（Ｏ、Ｓ－ＤＭＰ）．血中ＷＡＰ以乙、乙酸乙酯混合溶剂提取、以磷酸三丁酯为内标测定；尿液中Ｏ、Ｓ－二甲基磷酸酯（Ｏ、Ｓ－ＤＭＰ）以乙酸提取，以ＤＭＴＰ氘氢化合物为内标测定。ＭＡＰ最小检出量为１０ｎｇ／ｍｌ血，回收率为７５．１％，Ｏ、Ｓ－ＤＭＰ的最小检出量为２０ｎｇ／ｍｌ尿．     

RP-HPLC测定全血中三环类抗抑郁药的研究

本文建立了反相高效液相色谱法，测定全血中三环类抗抑郁药：阿米替林、多虑平、三甲丙咪嗪、氯丙咪嗪、丙咪嗪、去甲丙咪嗪。以乙酸苯胺为内标物。血样碱化后，用正己烷：异戊醇（９９∶１）提取，０．１ｍｏｌ／Ｌ硫酸及提。色谱柱为ＨｙｐｅｒｓｉｌＣｌｓ；流动相为乙腈：０．０１ｍｏｌ／Ｌ磷酸二氢钾（含０．０１ｍｏｌ／Ｌ二乙胺的水溶液并加磷酸调至ｐＨ３．０（３∶７）；２１０ｎｍ检测。线性范围为０．０１５６—２．０ｕｇ／ｍｌ，最低检出浓度为１０ｎｇ／ｍｌ（在全血中）。本法简便、快速，适用于法庭毒物分析及临床治疗药物监测。     

度冷丁滥用者尿中原体及其代谢产物的分析研究

本研究利用ＧＣ／ＭＳ（ＥＩ．ＰＣＩ）技术，在度冷丁滥用者尿中确认了６种代谢产物。并建立了ＧＣ／ＦＩＤ分析度冷丁及其代谢产物的方法。方法线性范围为０．１～２０μｇ／ｍｌ，变异系数小于１０％，最小检出量为５０ｎｇ／ｍｌ，适用于度冷丁成瘾者尿中含量的测定。     

度冷丁滥用者尿中原体及其代谢物产物的分析研究

本研究利用ＧＣ／ＭＳ（ＥＬＰＣＩ）技术，在度冷丁滥用者不中确认了６种代谢产物，并建立了ＧＣ／ＦＩＤ分析度冷丁及其代谢产物的方法，方法线性范围０．１～２０μｇ／ｍｌ，变异系数小于１０％，最小检出量为５０ｎｇ／ｍｌ，适用于度冷丁成瘾者尿中含量的测定。     

高效液相色谱法检测人脊髓听利多卡因

建立高效液相色谱法对人脊髓中利多卡因进行分析的方法，以扩大药（毒 ）物检测药围及检测手段，适应法医学鉴定及对特殊检材检验的需要，用利多卡因标准品及空白脊髓标准添加实验，对色谱条件、样品处理方法，回收率及方法的线性和精密度进行了系统考察。所建方法线性范围是２．０～２０．０μｇ／ｍｌ（ｒ＝０．９９９９），最低检测限为０．２μｇ／ｍｌ（Ｓ／Ｎ≥），加样回收率为８２．４％～９２．７％，该法选择性好、不受干     

大鼠闭合性脑损伤后血清髓鞘碱性蛋白研究

用ＥＬＩＳＡ双抗体夹心方法研究了大鼠闭合性弥漫性脑损伤后,血清髓鞘碱性蛋白（ＭＢＰ）含量的时序变化。正常大鼠血清中ＭＢＰ为６．１６３３±１．５３０１ｎｇ／ｍｌ（Ｘ±Ｓ）。伤后立即死亡组大鼠的血清ＭＢＰ为１１．３８１８±２．６５７４ｎｇ／ｍｌ,伤后１５ｍｉｎ,为１０．８３１９±２．３１３５ｎｇ／ｍｌ,此血清高ＭＢＰ水平一直持续到伤后３天。伤后第４天和第５天,血清ＭＢＰ基本恢复到正常水平。立即死亡组、伤后１５ｍｉｎ组和伤后３天之内各组的ＭＢＰ水平与正常组和伤后第４天和第５天的比较,Ｐ＜０．０１。认为可进一步探讨将血清ＭＢＰ含量的检测,作为脑震荡性脑损伤的辅助生化诊断指标之一     

酒精对乌头碱毒物动力学的影响

用反相高效液相色谱法测定生物样品中的乌头碱．给新西兰兔灌服１．５ｇ／ｋｇ短柄乌头后，乌头碱在免休内的毒物动力学过程为二室开放模型，其Ｋａ、Ａ、Ｂ值分别为１．１６２９±０．４０５３，０．６０４６±０．２５７４，１．１６０７±０．３７８１ｕｇ／ｍｌ．研究了灌服５．７５ｇ酒精对该动力学过程的影响，表明酒精不改变其模型类型，但可明显加速乌头碱的吸收和分布（Ｐ＜０．０１），对消除过程无明显影响，其Ｋａ、Ａ、Ｂ值分别为２．４０２６±０．５３７６，１．２０５１±０．５３２８，１．２０３７±０．４０９５μｇ／ｍｌ。说明酒精可增加短柄乌头的毒性。     

GC／MSD分析生物样品中的巴比妥类药物

本文建立了ＧＣ／ＭＳＤ分析生物样品中五种巴比妥类药物的测定方法。线性范围是２００～５０００ｎｇ／ｍｌ，方法的回收率：巴比妥８１．４１±４．５０；异戊巴比妥８３．５８±６．６５；戊巴比妥８２．７８±７．１７；司可巴比妥７９．０４±７．２０；苯巴比妥８２．１３±６．８１（ｎ＝６）。     

蛇毒检测方法的实验研究

目的建立系统的蛇毒检测方法。方法通过对3种常见蛇毒样品检测,确定用ELISA法检测蛇毒的最适条件。结果本研究所建立的ELISA蛇毒检测法灵敏度为3.9ng/ml;交叉实验结果显示,该方法特异性高,只有个别蛇毒存在交叉反应;样本从采集到检出时间在1h40min之内;动物实验显示,半致死量中毒家兔的血液蛇毒检测在中毒后48h内有效,中毒死亡家兔的血液蛇毒检测在死后72h内有效。结论所建立方法灵敏、准确、快速、简便,是一种可靠的蛇毒检测法。     

Rapid and simple GC-MS method for determination of psychotropic phenylalkylamine derivatives in nails using micro-pulverized extraction

A rapid and simple gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the simultaneous detection and quantification of five psychotropic phenylalkylamines (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, and norketamine) in toenails. After external decontamination, nail clippings were mechanically pulverized with a bead mill and then incubated in methanol under ultrasonication at 50°C for 1 h. The resulting solutions were evaporated to dryness, derivatized, and analyzed by GC-MS. The intra- and inter-day precisions were within 10.7% and 13.9%, respectively. The intra- and inter-day accuracies were -4.2% to 5.0% and -2.4% to 8.4%, respectively. Limits of detection and quantification for each analyte were lower than 0.024 and 0.08 ng/mg, respectively. The recoveries were in the range of 80.6-87.5%. The results indicated that the proposed method is a simple, rapid, accurate, and precise for quantification of five phenylalkylamines in nails. The method was successfully applied to the simultaneous detection and quantification of phenylalkylamines in nail samples of possible drug abusers.     

用ABC法鉴定眼镜蛇中毒死亡的实验研究

S-D 大白鼠皮下分别注射不同剂量的眼镜蛇粗毒,于死后不同时间取注射部位肌肉及肝、肾、心、肺等做冰冻切片和石蜡切片。用ABC 免疫组化法检测蛇毒,观察蛇毒的组织定位。在注射部位的骨骼肌内检出特异性染色的蛇毒。HE 染色各器官未见特征性改变。蛇毒量为2LD_(50) ,死亡24小时后取材仍有阳性反应。蛇毒多结合于注射部位的骨骼肌细胞膜上。注射11LD_(50) 的蛇毒后,发现肾小球毛细血管内皮细胞和肝细胞核上结合有蛇毒。家兔注射眼镜蛇毒或被眼镜蛇咬伤后,注射部位和咬伤部位肌肉中也可检出蛇毒。本实验首次使用ABC 免疫组化染色,取得了迄今为止直接在动物组织上对蛇毒定位检测的最好结果。     

Comparative studies on tissue distributions of organophosphorus, carbamate and organochlorine pesticides in decedents intoxicated with these chemicals

This paper describes the tissue distributions of dichlorvos, an organophosphate, chlorpyrifos-methyl, an organophosphorothioate, methomyl, a carbamate, and endrin, an organochlorine, in three individuals (Cases 1-3) who died after ingesting insecticidal preparations containing these chemicals. In Case 1 involving dichlorvos and chlorpyrifos-methyl, no dichlorvos was detected in most of the blood and tissue samples. Tiny amounts of dichlorvos (0.067 mg/L and 0.027 mg/L) were detected in the vitreous humor and cerebrospinal fluid, respectively. The chlorpyrifos-methyl concentrations in the blood samples were very site-dependent with a range of 0.615-2.24 mg/L. The tissue concentrations of chlorpyrifos-methyl were within the range 0.379-8.60 mg/kg. The total amounts of dichlorvos and chlorpyrifos-methyl in the stomach were 879 and 612 mg, respectively. The serum cholinesterase activity was 3 IU/L/37 degrees C. In Case 2 involving methomyl, the methomyl concentrations in the blood samples were very site-dependent with a range of 0.56-4.75 mg/L. The tissue concentrations of methomyl were 2.61 mg/kg or less, no methomyl being detected in the spleen, liver and kidney. The methomyl concentrations in the cerebrospinal fluid and vitreous humor were 5.37 and 4.75 mg/L, respectively. The stomach contained 85 mg methomyl. The serum cholinesterase activity was 73 IU/L/37 degrees C. In Case 3 involving endrin, the victim underwent medical treatment for 7 h after ingesting an endrin preparation. The differences in the endrin concentrations among the blood samples were small, with a range of 0.353-0.615 mg/L. The tissue concentrations of endrin were within the range 0.467-13.3 mg/kg. The endrin in the stomach (66 mg) was adsorbed almost completely on the activated charcoal that was administered for medical treatment.     

Species identification from dried snake venom

Illegal trade in snake parts has increased enormously. In spite of strict protection under wildlife act, a large number of snakes are being killed ruthlessly in India for venom and skin. Here, an interesting case involving confiscation of crystallized dried snake venom and subsequent DNA-based species identification is reported. The analysis using the universal primers for cytochrome b region of the mitochondrial DNA revealed that the venom was extracted from an Indian cobra (Naja naja). On the basis of this report, the forwarding authority booked a case in the court of law against the accused for illegal hunting of an endangered venomous snake and smuggling of snake venom. This approach thus has immense potential for rapid identification of snake species facing endangerment because of illegal trade. This is also the first report of DNA isolation from dried snake venom for species identification.     

超分子溶剂萃取-气相色谱-串联质谱法检测尿液中苯二氮䓬类药物

目的建立一种尿液中9种苯二氮?类药物的超分子溶剂样品气相色谱-串联质谱(gas chromatography-tandem mass spectrometry,GC-MS/MS)分析方法。方法含9种苯二氮?类药物对照品的尿液样品用四氢呋喃和1-己醇组成的超分子溶剂进行液液萃取,取溶剂层氮吹至干,残余物用甲醇复溶后进行GC-MS/MS分析,数据采集方式为多反应监测模式,采用内标法定量。结果尿液中地西泮、咪达唑仑、氟硝西泮和氯氮平质量浓度在1~100ng/mL,劳拉西泮和阿普唑仑质量浓度在5~100ng/mL,硝西泮和氯硝西泮质量浓度在2~100ng/mL,艾司唑仑在质量浓度0.2~100ng/mL范围内具有良好的线性关系,相关系数为0.9991~0.9999,定量下限为0.2~5ng/mL,提取回收率为81.12%~99.52%,日内精密度[相对标准偏差(relative standard deviation,RSD)]和准确度(偏倚)分别小于9.86%、9.51%;日间精密度(RSD)和准确度(偏倚)分别小于8.74%、9.98%。室温和-20℃条件下,尿液中9种药物在15d内具有良好的稳定性。8名志愿者单摄口服阿普唑仑片后,在8~72h内尿液中阿普唑仑的质量浓度为6.54~88.28ng/mL。结论本研究建立的尿液中9种苯二氮?类药物的超分子溶剂萃取-GC-MS/MS分析方法,简便、快速、准确、灵敏,可为临床治疗及司法鉴定中苯二氮?类药物中毒监测提供技术支持。     

High performance liquid chromatographic determination of alcohols with reference to body distribution of methanol.

A method has been developed on reverse phase high performance liquid chromatography for simultaneous determination of methyl, ethyl and isopropyl alcohols under refractive index detection using pure water as the mobile phase. A good separation has been achieved between these alcohols. Detector response was linear with a detection limit of 5 mg/100 ml. Recovery studies were performed by adding known amounts of methyl and ethyl alcohols to blood, lung and liver within the range 80-90%. The reproducibility of the results was always greater than 90%. The quantitative distribution of methyl alcohol in postmortem body tissues and fluids has been reported in three cases of poisoning.     

Concentration-time profiles of ethanol in capillary blood after ingestion of beer.

Blood ethanol profiles were determined in experiments with healthy volunteers after they had drunk beer. When 330 ml of light beer (1.8% w/v ethanol) was consumed in 5 min by four men and four women, the average peak blood-alcohol concentration (BAC) reached was 8 mg/100 ml (range 2-11). After nine men had drunk 660 ml of beer (3.0% w/v or 3.6% w/v ethanol) in 25 minutes on an empty stomach, the average peak BAC was 32 mg/100 ml (range 26-44) and 37 mg/100 ml (range 23-54) respectively. When the same two beers were consumed by another nine men together with a meal, the peak BAC was 24 mg/100 ml (range 20-29) and 28 mg/100 ml (range 20-39) respectively. The peak BAC occurred earlier when beer was ingested together with food; mean 32 min (range 30-50) compared with 41 min (range 30-70) with an empty stomach. The rate of disappearance of alcohol from blood (beta-slope) was 12 mg/100 ml/h in the fed state and 15 mg/100 ml/h when subjects were fasted. The apparent volume of distribution of ethanol (Vd) was 0.65 l/kg (SD 0.07) for the empty stomach condition but exceeded unity when beer was ingested together with food. It seems that part of the dose of alcohol when consumed with food never reaches the systemic circulation.     

Dynamic changes of serum protein in rats with acute intoxication of Chinese cobra snake venom by proteomic analysis

To elucidate the toxic mechanism of snake venom at the protein level, proteomics technology was applied to investigate the effect of venom on circulation in the mammalian body. Temporal proteomic analysis was performed to profile the dynamic changes in the sera of Sprague–Dawley rats administered with Chinese cobra venom or saline. Using 8-plex iTRAQ analysis, 392 and 636 serum proteins were identified to be linearly upregulated or downregulated over time in the low-dose group and high-dose group, respectively. These proteins were mainly associated with the acute phase response pathway, complement system, and liver X receptor (LXR)/retinoid X receptor (RXR) and farnesoid X receptor (FXR)/RXR activation pathways. Compared with the low-dose group, the immune response and integrin pathways were inhibited in the high-dose group, although no obvious effect was observed. With consistently higher or lower expression in the high-dose group compared to the low-dose group throughout the whole process of venom poisoning, two proteins, Kininogen-1 (KNG1) and orosomucoid 1 (ORM1), which are involved in metabolism and immune response, occupied a core position in the pathway network and are considered venom dose-dependent biomarker candidates.     

Improved gas chromatography-negative ion chemical ionization tandem mass spectrometric method for determination of 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid in hair using mechanical pulverization and bead-assisted liquid-liquid extraction

A gas chromatography-negative ion chemical ionization tandem mass spectrometric (GC-NCI-MS/MS) method was developed and validated for the determination of 11-nor-Δ(9)-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) in human hair. After decontamination, hair samples were weighed (25mg), mechanically pulverized with a bead mill, and incubated in 0.7 mL of 1.0M sodium hydroxide at 95 °C for 30 min. Bead-assisted liquid-liquid extraction was performed with n-hexane:ethyl acetate (9:1, v/v), a method developed in our laboratory. The extract was evaporated to dryness, derivatized with pentafluoropropanol and pentafluoropropionic anhydride, and analyzed by GC-MS/MS in the negative ion chemical ionization mode using methane as the reagent gas. The linear ranges were 0.05-10.0 pg/mg for THC-COOH with the coefficient of determination (r(2) = 0.9976). The intra-day and inter-day precisions were within 1.7 and 13.8%, respectively. The intra-day and inter-day accuracies were -4.8 to 10.0% and -3.9 to 3.8%, respectively. The limit of detection and quantification were 0.015 and 0.05 pg/mg, respectively. The recoveries were in the range of 79.4-87.1%. The results indicate that the proposed method is simple, rapid, accurate, and precise for determination of THC-COOH in hair. The method identified THC-COOH in hair specimens from suspected marijuana abusers.