Miniaturized sample preparation method for determination of amphetamines in urine

A simple and miniaturized sample preparation method for determination of amphetamines in urine was developed using on-column derivatization and gas chromatography-mass spectrometry (GC-MS). Urine was directly applied to the extraction column that was pre-packed with Extrelut and sodium carbonate. Amphetamine (AP) and methamphetamine (MA) in urine were adsorbed on the surface of Extrelut. AP and MA were then converted to a free base and derivatized to N-propoxycarbonyl derivatives using propylchloroformate on the column. Pentadeuterated MA was used as an internal standard. The recoveries of AP and MA from urine were 100 and 102%, respectively. The calibration curves showed linearity in the range of 0.50-50 microg/mL for AP and MA in urine. When urine samples containing two different concentrations (0.50 and 5.0 microg/mL) of AP and MA were determined, the intra-day and inter-day coefficients of variation were 1.4-7.7%. This method was applied to 14 medico-legal cases of MA intoxication. The results were compared and a good agreement was obtained with a HPLC method.     

Gas chromatographic determination of cresols in the biological fluids of a non-fatal case of cresol intoxication

A simple and rapid method for analysis of free and conjugated cresols in biological fluids was developed. Prior to and following freeing of the conjugated cresols by acid hydrolysis in a sealed ampoule, free cresols were extracted by Extrelut column extraction, determined by gas chromatography, and confirmed by gas chromatography-mass spectrometry. In a non-fatal case of cresol intoxication a 46-year-old male had ingested about 100 ml of a saponated cresol soap solution. The concentrations of xylenol (2,4- and/or 2,5-dimethylphenol) and p- and m-cresol in the serum sample collected on admission were 15.8 micrograms/g, 43.3 micrograms/g and 73.8 micrograms/g, respectively. The total cresol concentration of 117 micrograms/g in the serum is within the range of fatal concentrations, and it is suspected therefore that the patient's recovery was due to adequate therapy alone.     

Toxicological study on intravenous thiopental anesthesia--interrelation between rate of injection and distribution of thiopental

A simple, rapid and accurate method to extract and clean thiopental in biological materials rich in fat was developed by using a separation column packed with Extrelut and Florisil. A Quantitative determination of thiopental by means of a gas chromatograph equipped with a flame photometric sulfur detector was attempted. The calibration curve was linear over the range of 5-200 micrograms/ml. Extraction of replicate tissues involving 3 micrograms of thiopental resulted in a recovery of the 99.7-101.8% range and a coefficient of deviation of the 0.3-1.9% range. Replicate extraction of five tissues from rats treated with thiopental resulted in a coefficient of deviation of below 6%. In rats given 40 mg/kg of thiopental and sacrificed after 5 min, the thiopental levels in fat were found to be higher after a slow intravenous injection than after a quick injection. The same tendency, however, was not observed in other tissues. It seems that the rate of intravenous thiopental injections might be estimated by comparison with thiopental levels in fat tissues.     

On-column derivatization for determination of amphetamine and methamphetamine in human blood by gas chromatography-mass spectrometry

A simple determination method of amphetamine (AP) and methamphetamine (MA) in human blood was developed using on-column derivatization and gas chromatography-mass spectrometry (GC-MS). AP and MA were adsorbed on the surface of Extrelut and then derivatized the N-propoxycarbonyl derivatives using propylchloroformate. Pentadeuterated MA was used as an internal standards. The recoveries of AP and MA from the spiked blood were 89.7 and 90.3%, respectively. The calibration curves showed linearity in the range of 12.5-2000 ng/g for AP and MA in blood. The coefficients of variation of intraday and interday were 0.42-4.58%. Furthermore, this proposed method was applied to some medico-legal cases of MA intoxication. MA and its metabolite AP were detected in the blood samples, and the correlation of the blood level of amphetamines and the behaviors of the victims was in good agreement with the criteria proposed by Nagata [Jpn. J. Legal Med. 37 (1983) 513].     

Determination of disulfoton and its metabolites in the body fluids of a Di-Syston intoxication case

Disulfoton and its metabolites, two sulfoxides and two sulfones, in the body fluids of a patient who had ingested Di-Syston were analyzed by FPD-GC and GC/MS. After the chemicals in the extract (Fraction 1) obtained by Extrelut column extraction were analyzed, disulfoton and sulfoxides in Fraction 1 were oxidized into sulfones. The sulfones in the extract (Fraction 2) obtained by Extrelut column extraction were analyzed and the estimated concentrations of metabolite were calculated. The concentrations of disulfoton and the sum of the metabolites in the blood collected on admission were 0.093 nmol/g (25.4 ng/g) and 4.92 nmol/g (corresponding to 1.35 micrograms/g of disulfoton), respectively. These concentrations appear to indicate a severe level of disulfoton intoxication.     

固相微萃取-气相色谱质谱法测定血浆中的氯氮平浓度

目的建立固相微萃取-气相色谱质谱法测定人血浆中氯氮平浓度的方法。方法以固相微萃取法提取血浆中的氯氮平,萃取头为100μm聚二甲基硅氧烷,洛沙平作内标,用气相色谱质谱选择离子法进行检测。结果本文建立的方法在5~2000 ng/ml浓度范围内呈线性关系,检测限为0.1 ng/ml(信噪比>3),低、中、高浓度(100、500、1000 ng/ml)平均相对回收率分别为98.6%、94.6%和94.6%,日内、日间RSD分别小于7.4%和7.1%。结论本文建立的固相微萃取-气相色谱质谱法灵敏度高、准确度好、操作简便,适用于氯氮平急性中毒案件的检测。     

GC法检测血液和尿液中甲基苯丙胺和咖啡因

目的建立同时测定血、尿中甲基苯丙胺和咖啡因含量的方法。方法应用GC／NPD技术,以4-苯基丁胺为内标,直接碱化,用氯仿提取,三氟乙酸酐衍生化,8CB熔融石英毛细管柱（30m×0．25mm×0．25μm）分析。结果生物样品中甲基苯丙胺与咖啡因在0．012—7．5μg/mL浓度范围内线性关系良好,检测限（S／N=3）依次为1．2ng／mL,0．6ng／mL（血）;1．6ng／mL,0．8ng／mL（尿）。苯丙胺在0．017—10．0μg／mL浓度范围内线性关系良好,检测限为1．6mg／mL（血）,3．2ng／mL（尿）。所有样本回收率均大于85％。结论本方法准确、灵敏,适用于血、尿中甲基苯丙胺及其代谢物苯丙胺的三氟乙酸酐衍生化物和咖啡因的同时检测,为判定滥用毒品种类、追查毒品来源以及研究生物体内甲基苯丙胺和咖啡因的交互影响提供了检测手段。     

Headspace solid phase microextraction for the gas chromatographic analysis of methyl-parathion in post-mortem human samples. Application in a suicide case by intravenous injection

A simple and rapid procedure for the determination of methyl-parathion (m-p) in post-mortem biological samples was developed using headspace solid phase microextraction (SPME) and gas chromatography (GC) with nitrogen-phosphorous detection (NPD). Methyl-parathion was extracted on 85 microm polyacrylate SPME fiber. Salt addition, extraction temperature, and extraction time were optimized to enhance the sensitivity of the method. The linearity (y = 0.0473x - 0.0113, R2 = 0.9992) and the dynamic range (0.1-40 microg/ml) were found very satisfactory. The recoveries of methyl-parathion were found to be 46% in spiked human whole blood, 53% in spiked homogenized liver tissue, and 54% in spiked homogenized kidney tissue compared with samples prepared in water. The coefficients of variations for 2, 4, and 20 microg/ml of methyl-parathion in blood ranged from 0.9 to 5.1%, whereas the detection limit of the method was satisfactory (1 ng/ml in aqueous samples, 50 ng/ml in whole blood). The developed procedure was applied to post-mortem biological samples from a 21-year-old woman fatally poisoned (suicide) by intravenous injection of methyl-parathion. The intact insecticide was found in the post-mortem blood at a concentration of 24 microg/ml. No methyl-parathion was detected in the liver, kidneys, and gastric contents.     

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。     

Detection of S-methylfenitrothion, aminofenitrothion, aminofenitroxon and acetylaminofenitroxon in the urine of a fenitrothion intoxication case

A 23-year-old male attempted suicide by ingesting approximately 50 ml of 5% fenitrothion emulsion, and vomited soon afterwards. He was admitted to a hospital about 3 h after ingestion. He recovered and was discharged from hospital 3 days after admission. The serum cholinesterase activity (normal range: 175-440 I.U.) was only 29 at 3 h, 32 at 1 day, 59 at 2 days and 75 at 3 days after ingestion. Fenitrothion and its metabolites in the body fluids were extracted by an Extrelut column extraction method, detected by a gas chromatograph equipped with either a hydrogen flame ionization detector or a flame photometric detector, and confirmed by a gas chromatograph-mass spectrometer. Fenitrothion concentration in the blood was 169.5 ng/g at 3 h after ingestion. The half life of blood fenitrothion concentration was found to be about 4.5 h. Fenitrothion metabolites, 3-methyl-4-nitrophenol, aminofenitrothion, aminofenitroxon, acetylaminofenitroxon and S-methylfenitrothion, were detected in the urine samples. All of them except S-methylfenitrothion were detected in the urine samples collected up to 62 h after ingestion. It would appear therefore that fenitrothion poisoning can be determined by detection and analysis of the metabolites in urine even if fenitrothion has not been detected in the blood.     

体液中常见滥用药物的系统筛选分析

本文建立了体液中常见滥用药物的筛选分析体系.尿液或血液经固相萃取(SPE)或液提取(LLE)后,直接用GC/NPD分析或经TFA、BSTFA衍生化后用GC/MS分析.方法适用于同时分析甲基苯丙胺、MDMA、度冷丁、去甲度冷丁、曲马多、美沙酮、EDDP、可卡因、苯甲酰芽子碱、可待因、安定、氯丙嗪、吗啡、单乙酰吗啡等十四种常见滥用药物及代谢物.SPE法和LLE法回收率分别为66～102%和50～86%,最低检出限为2-5ng/ml尿.涉毒案件的鉴定应用表明该分析方法简便、快速、可靠.     

A comparative analysis of Cannabis material

Extracts of 100 plant-like or resinous materials were analyzed for CBD, CBC, delta 9-THC, and CBN by GC using two different column packings and by GC-MS. Our independent identification of these cannabinoids confirmed those of other forensic science analysts who used microscopic examination, the Duquenois-Levine color test, and TLC for their analyses of the same samples. The identifications of cannabinoids by forensic acience analysts using TLC were corroborated by GC-MS analysis of hexane extracts of appropriate chromatogram spots.     

用Oasis^TM固相柱研究检测腐败生物组织中的吗啡

目的建立一种适合大批量腐败生物检材的标准化分析测定方法。方法应用新型亲水亲酯的OasisTM固相柱,并使用Zymark全自动固相萃取仪,对腐败生物组织中的吗啡进行检测。结果与传统的C18固相柱相比较,OasisTM固相柱具有选择范围广、吸附能力强的特点,其方法操作简单、快速、重现性好及回收率高。结论该方法可应用于案件的检测工作。     

人血、尿中富马酸喹硫平的气相色谱分析

目的建立人血、尿中富马酸喹硫平的气相色谱分析方法。方法用乙醚提取血、尿中的富马酸喹硫平，直接对其进行定性、定量分析。以正常人血、尿为空白样本，分别添加标准富马酸喹硫平，确定检材的前处理方法、色谱分析条件、工作曲线、线性范围、方法的精密度、回收率等，并对1例大剂量服用富马酸喹硫平中毒死者的体液浓度进行测定。结果该方法分析血、尿中富马酸喹硫平的线性范围分别为8．0～800．0μg／ml和20．0—800．0μg/ml；最低检测限分别为0．04μg／ml和0．10μg／ml（S／N≥3），日内、日间精密度均小于4％，回收率在97．08％-101．42％之间。结论该分析方法操作便捷、实用、准确度高，适用于富马酸喹硫平的临床血药浓度快速监测和法医毒物鉴定。     

Practical GC/MS analysis of oxidation dye components in hair fiber as a forensic investigative procedure

The purpose of this study was to improve the reliability of discrimination (or identification) of dyed hair by analyzing chemical substances present in the hair, as an addition to the conventional macroscopical and microscopical examinations and ABO blood group examination. Oxidation hair-dye components such as p-phenylenediamine (PPDA), toluylene-2,5-diamine (T-2,5-DA), o-aminophenol (OAP), m-aminophenol (MAP), p-aminophenol (PAP) and p-amino-o-cresol (PAOC) were selected as the object of study. After alkaline-digestion, hair samples were adjusted to a pH of 12.6 to 12.8, and applied onto an Extrelut column. After 15 min, the components were simultaneously extracted and derivatized with n-hexane including 1% heptafluoro-n-butyryl (HFB) chloride. Their HFB derivatives within a condensed sample were diluted in ethyl acetate, and analyzed by gas chromatography-mass spectrometry (GC-MS) with full mass scanning or selected ion monitoring. For estimating their levels, 2,4,6-trimethylaniline was used as the internal standard. Standard curves obtained by preparing a 5 cm segment of control hair spiked with authentic substances were linear, ranging from 0.1 to 4.0 micrograms for PPDA and T-2,5-DA, and from 0.01 to 0.4 microgram for OAP, MAP, PAP and PAOC. The coefficient of variation of inter-day precisions (each n = 4) was below 16% for PPDA, below 20% for OAP and PAP and below 24% for T-2,5-DA, MAP and PAOC. These components were detectable even at 6 ng for PPDA, T-2,5-DA, MAP and PAP, 8 ng for OAP, and 4 ng for PAOC. Recovery percents using this procedure ranged from 54 to 86%. By using actual dyed hair samples this method was shown to provide high sensitivity for accurate detection.     

血尿肝中毒鼠强硅藻土提取GC／NPD检测法

目的建立硅藻土提取气相色谱测定血、尿、肝中毒鼠强的方法。方法原尿液、血液用水稀释、肝匀浆用6％高氯酸沉淀蛋白的上清液倒入硅藻土小柱中，血和尿用苯洗脱，肝用三氯甲烷洗脱，挥干洗脱液，用甲醇定容至0．1ml。结果血提取率98．4％，尿提取率95．6%，肝提取率98．1％。相对标准偏差低于3．2％，检出限低于20ng／ml（g）。结论该法简便、快速，提取率高，适合作为常规毒物分析方法。     

LC／MS／MS法测定全血中马钱子碱和士的宁的含量

目的建立定量分析人全血中马钱子碱和士的宁的高效液相色谱质谱联用法。方法应用Oasis^TM MCX小柱进行固相萃取法提取，XTerra^TM RP18色谱柱分离。结果在该条件下，人血中马钱子碱和士的宁的线性范围为0．01—5．0μg／ml，最小检出限为0．2ng／ml；马钱子碱和士的宁在0．01—5．0μg／ml浓度范围内的回收率均在80％以上。结论高效液相色谱质谱联用法可定量测定血中马钱子碱和士的宁。     

Fatal colchicine poisoning

In two cases, after i.v. injections of colchicine serum concentrations of 170 ng/ml and 240 ng/ml were found before death. Colchicine was extracted from serum with the aid of Extrelut extraction columns and analysed using HPLC. Urine was also analysed and the findings were confirmed by drawing a UV-curve for colchicine using the stop-flow method. In one case, colchicine was still detectable in urine 10 days after injection.     

One-step liquid-liquid extraction of cocaine from urine samples for gas chromatographic analysis

An improved technique for cocaine extraction from urine samples for gas chromatographic (GC) analysis is described. Employing a simple liquid-liquid extraction (LLE) of cocaine with a mixture of ethyl ether:isopropanol (9:1) the method presents a mean recovery of 74.49%. Limit of detection (LOD) and limit of quantification (LOQ) were 5 and 20 ng/ml, respectively. The method is highly precise (coefficient of variation (CV) <8%) and linear from 20 to 2000 ng/ml. It can he applied to detect the presence of cocaine in urine as a marker of its recent use in drug abuse treatment protocols.     

A fatal case of oleandrin poisoning

The study presents a case of fatal poisoning with oleander leaves in an adult diabetic male. After repeated vomiting, and gastrointestinal distress the patient was admitted at the hospital with cardiac symptoms 1h after the ingestion. Urine samples were assayed immunochemically and by GC-MS for drugs of abuse and for general toxicological screen. Blood was analyzed for alcohol and volatiles by static head space GC-MS. Blood and oleander leaves were analyzed by LC-MS/MS for oleandrin and related compounds, the main cardiac glycosides of Nerium oleander. Oleandrin was detected by LC-MS/MS in the blood sample at a concentration of approximately 10 ng/ml. Another cardiac glycoside with pseudo-molecular ion of m/z 577, a likely structural isomer of oleandrin, was also detected in the blood and oleander leaves. However, by using the response as a function of concentration for oleandrin, this cardiac glycoside was roughly estimated at a concentration of approximately 10 ng/ml in the deceased blood. This would give a total fatal blood concentration of cardiac glycosides of about approximately 20 ng/ml in the deceased blood.