Simple and simultaneous quantification of cyanide,ethanol, and 1-propanol in blood by headspace GC–MS/NPD with Deans switch dual detector system

Cyanide is a powerful and rapidly acting poison. In Japan, cyanide poisoning is rare, and regular cyanide testing can be costly and time consuming. In contrast, alcohol analysis is routinely performed in most forensic laboratories. In this study, we attempted to develop a method for the simultaneous quantification of cyanide and alcohols in blood using headspace gas chromatography (HS–GC). As nitrogen-phosphorus detection (NPD) is more sensitive to hydrogen cyanide than mass spectrometry (MS), a Deans switch was used to switch the detectors during a single run. The separation provided by three analytical columns, PoraBOND Q, CP-Sil 5 CB, and HP-INNOWax, was investigated, and PoraBOND Q was selected. The use of HS–GC–MS/NPD with a Deans switch enabled the simple and simultaneous quantification of cyanide, ethanol, and 1-propanol. Eighteen other volatile compounds were detected in the SIM/scan mode of the MS.     

HS-GC／FID分析血中的乙醚

目的建立顶空GC/FID分析血中乙醚的方法。方法用顶空GC/FID分析血中乙醚,并对萃取条件和色谱条件进行优化。结果方法的检出限为0.3ug/ml,回收率为81%,相对标准偏差为4.4%。结论该法简便、快速、灵敏,适合毒物分析检验的要求。     

Highly sensitive analysis of methamphetamine and amphetamine in human whole blood using headspace solid-phase microextraction and gas chromatography-mass spectrometry

A simple and highly sensitive method for analysis of derivatized methamphetamine (MA) and amphetamine (AM) in whole blood was developed using headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry electron impact ionization selected ion monitoring (GC-MS-EI-SIM). A whole blood sample, deuterated-MA (d(5)-MA), as an internal standard (IS), tri-n-propylamine and pentafluorobenzyl bromide were placed in a vial. The vial was heated and stirred at 90 degrees C for 30min. Then the extraction fiber of the SPME was exposed at 90 degrees C for 30min in the headspace of the vial while being stirred. The derivatives adsorbed on the fiber were desorbed by exposing the fiber in the injection port of a GC-MS. The calibration curves showed linearity in the range of 0.5-1000ng/g for both MA and AM. The time for analysis was about 80min per sample. In addition, this proposed method was applied to two autopsy cases where MA ingestion was suspected. In one case, MA and AM concentrations in the mixed left and right heart blood were 165 and 36.9ng/g, respectively. In the other case, MA and AM concentrations were 1.79 and 0.119 microg/g in the left heart blood, and 1.27 and 0.074 microg/g in the right heart blood, respectively.     

Fatal and non-fatal methomyl intoxication in an attempted double suicide

A 79-year-old man and his 73-year-old wife attempted double suicide by ingesting methomyl powder. The woman died 19 h after ingestion in spite of intensive care. At autopsy a large number of miliary hemorrhages were found in both thalami of the brain. Her husband, however, recovered after 10 days of treatment. Methomyl (CAS No. 16752-77-5, Lannate) in the biological materials was analyzed by gas chromatography-mass spectrometry. The methomyl concentration was 44 micrograms/g in the wife's serum sample collected 1 h after ingestion, and 0.2 microgram/g in the blood sample collected at autopsy. The methomyl concentration in the husband's blood sample collected 28 h after ingestion was from 0.01 to 0.1 microgram/g. It is suggested that prompt and adequate intensive care including a direct hemoperfusion is necessary to effect the recovery of patients with lethal blood levels of methomyl. The miliary hemorrhages found in the thalami of the brain are suspected to have been caused by asphyxia induced by methomyl intoxication.     

Detection of the synthetic drug 4-fluoroamphetamine (4-FA) in serum and urine

4-Fluoroamphetamine (4-FA) was detected in the blood and urine of two individuals suspected for driving under the influence (DUI). The test for amphetamines in urine subjected to immunoassay screening using the CEDIA DAU assay proved positive. Further investigations revealed a 4-FA cross-reactivity of about 6% in the CEDIA amphetamine assay. 4-FA was qualitatively detected in a general unknown screening for drugs using GC/MS in full scan mode. No other drugs or fluorinated phenethylamines were detected. A validated GC/MS method was established in SIM mode for serum analysis of 4-FA with a limit of detection (LOD) of 1 ng/mL and a lower limit of quantification (LLOQ) of 5 ng/mL. Intra-assay precision was approx. 4% and inter-assay precision approx. 8%. Applying this method, the 4-FA serum concentrations of the two subjects were determined to be 350 ng/mL and 475 ng/mL, respectively. Given the pharmacological data of amphetamine, 4-FA psychoactive effects are to be expected at these serum levels. Both subjects exhibited sympathomimetic effects and psychostimulant-like impairment accordingly.     

GC/MS法联合分析生物检材中的尼美舒利、二氧丙嗪

目的建立生物检材中尼美舒利、二氧丙嗪的GC/MS检验方法。方法采用浸取法提取,GC/MS定性检验,内标法选择离子检测（SIM）模式定量检验。结果最低回收率为69%,尼美舒利的线性相关系数为0.998,克咳敏的线性相关系数为0.9973。结论该方法操作简便快速,提取率高,可用于实际案件中尼美舒利、二氧丙嗪的检验。     

Accidental autoerotic death by volatile substance abuse or nonsexually motivated accidents?

In contrast to typical autoerotic fatalities, when death is due to asphyxia mostly by mechanical compression of the neck, atypical autoerotic accidental deaths (AADs) involve sexual self-stimulation by other means such as electrocution or inhalation of chemical agents. Especially in lethal cases of volatile substance abuse (VSA), a differentiation between suicide or sexually or nonsexually motivated accident is often complicated in practical casework. Considering the small number of AADs involving chemical substance abuse reported in the literature, the number of unreported cases seems to be very high. We report about 5 lethal cases of VSA; analysis was performed using headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC/MS). When headspace sampling is not performed at autopsy, the analysis of volatile substances can be very complicated. In 2 cases, an AAD was diagnosed considering findings at the scene, reconstruction of the event, and discussion of the circumstances of the death. These findings demonstrate the importance of VSA in atypical autoerotic asphyxia. Therefore, in cases of suspected lethal inhalational intoxications, as a matter of principle, headspace asservation should be performed at autopsy and an autoerotic motivational background should be taken into consideration for differential diagnosis.     

顶空GC／MS法检验血液中的硫化氢

目的建立血液中硫化氢的气相色谱质谱联用分析方法。方法取心血3mL-5mL,置于20mL顶空瓶中,加入1g氯化钠,加3mL-5mL蒸馏水,加入2mol／L盐酸1mL,加盖密封,混匀后于80℃水浴中加热20min。取液上气体0．5mL进样分析。结果在中毒死亡者的心血中检出硫化氢,保留时间参考值为3．6min。结论该方法可用于刑事案件中硫化氢的快速分析。     

氟乙酸五氟苄基酯的气相色谱分析

目的 对氟乙酸根阴离子的衍生物-氟乙酸五氟苄基酯(PFB-MFA)进行了气相色谱分析，以满足氟乙酰胺类鼠药化验工作的需要。方法 使用GC／ECD、GC／MS Scan、GC／MS SIM对该化合物的标准溶液进行了分析，得到了各种分析方法的校正曲线及检测限。结果 三种方法测定PFB-MFA的线性范围分别为GC／ECD 0.01～0.1ng／μl、GC／MS SCAN 1～100ng／μl、GC／MS SIM 5×10~(-3)～1ng／μl。三种方法对氟乙酸根阴离子的检测限分别为：GC／ECD1.31×10~(-4)ng／μl、GC／MS SCAN 0.13ng／μl、GC／MS SIM 1.76×10~(-4)ng／μl。结论 氟乙酸根阴离子的衍生物氟乙酸五氟苄基酯在GC／ECD及GC／MS上具有一定的线性范围，有非常高的灵敏度，可用于法庭科学的定性定量分析。     

Evaluation of the Abbott TDx-radiative energy attenuation (REA) ethanol assay in a study of 1105 forensic whole blood specimens

In a preliminary study to determine the applicability of the Abbott radiative energy attenuation (REA) method for the quantification of ethanol in whole blood specimens it was concluded that a larger number of samples was required to evaluate the method, particularly for use in forensic toxicology applications. In this study, 573 blood specimens from suspected driving while intoxicated individuals (DWI blood) and 532 postmortem blood specimens (PM blood) were analyzed by the REA method and a headspace gas chromatographic method (GC) currently used in this laboratory. "Negative" specimens (less than 10 mg/dL by GC) and "positive" specimens (greater than or equal to 10 mg/dL by GC) in each category were analyzed. Linear regression analysis comparing the REA values with the GC values was performed for each type of blood specimen. The equation obtained for DWI blood specimens was REA = 0.943 GC + 1.54; the equation for PM blood specimens was REA = 0.980 GC + 2.76. The correlation coefficient for each group was greater than 0.99. The data suggested that a limit of detection of 10 mg/dL could be applied for DWI blood specimens, while 20 mg/dL would be recommended as the limit of detection for PM blood specimens.     

GC／MS检测尿中4种苯氧羧酸类除草剂

目的建立尿中2，4-D、2，4-DP、MCPA、MCPP等4种苯氧羧酸类除草剂的分析方法；方法液液提取分离，乙醚为提取液，DCPA为内标，硫酸正丁醇酯化衍生化，气质联用分析法分析；结果尿中4种除草剂添加样品的相对回收率在80％以上，检测限都在5ng／ml以下，对中毒兔尿样进行了分析；结论对实际发生的中毒案件分析有足够的灵敏度。     

Examination of the effect of dose-death interval on detection of meperidine exposure in decomposed skeletal tissues using microwave-assisted extraction

The effect of dose-death interval and tissue distribution on the detection of meperidine in selected skeletal tissues was examined using a rapid microwave-assisted extraction (MAE) methodology. Rats (n=14) were dosed with 0 (n=2) or 30 mg/kg (n=12) meperidine (i.p.). Drug-positive rats were sacrificed with CO(2) after 20, 30, 90 and 150 min (n=3 per group). Heart blood was collected immediately after death. Tibiae were excised and frozen for further analysis. The remaining carcasses were allowed to decompose outside in secured cages to the point of complete skeletonization in a rural Northern Ontario location during the late summer months. Vertebrae and pelvi were collected for each animal. Tibial marrow was homogenized in 3 mL PB6 (phosphate buffer, 0.1M, pH 6). Fresh tibiae, and decomposed vertebrae and pelvi were cleaned in PB8.5 (phosphate buffer, 0.1M, pH 8.5) and sonicated to remove remaining soft tissue. Samples of dried, ground bone (0.5-1g) suspended in 2 mL PB6 were then irradiated in a domestic microwave oven (1100 W) at atmospheric pressure for 15 min. Samples of vertebral bone (1g) were also extracted by passive incubation in methanol (3 mL, 50°C, 72 h). All supernatants then underwent solid-phase extraction and analysis by GC/MS, using electron impact ionization in the Selected Ion Monitoring (SIM) mode. Mean GC/MS responses for each tissue type were negatively correlated with dose-death interval, with correlation coefficients ranging from -0.32 to -0.87. Analysis of variance showed dose-death interval to be a main effect (p<0.05) with respect to GC/MS response for blood, marrow, tibial epiphyses prepared by MAE, and vertebral bone prepared by passive extraction, but not for tibial diaphyses, pelvi or vertebrae prepared by MAE. Overall, MAE is advantageous as a rapid extraction tool for screening purposes in skeletal tissues, but assignment of significance to quantitative expressions of skeletal drug concentrations is complex and should be approached with caution.     

Quantitative analysis of chlorpromazine by electron spin resonance (ESR) spectroscopy.

A simple and sensitive method is described for quantitative analysis of chlorpromazine in blood, serum, urine and tissue homogenate. The chlorpromazine cation radical produced by adding perchloric acid and 2,3-dichloro-5,6-dicyano-p-benzoquinone to the sample can be detected by the ESR method at room temperature. The sensitivity limit is 10 ng, that is, 20 microliters of the solution containing 0.5 microgram chlorpromazine/ml. The time needed for the measurement is within 10 min. The chlorpromazine radical thus produced is very stable; for example, 95% of the radical was observed after 24 h. The advantage of this method is discussed by comparing with the ordinary spectrophotometry which requires the purification of the sample.     

分散固相萃取-GC/MS法测定人血中啶虫脒的含量

目的建立人血中新型农药啶虫脒的检验方法。方法用分散固相萃取法提取人血中啶虫脒,GC/MS/SIM法进行测定。结果在0.02μg/mL～2μg/mL添加水平范围内,线性关系良好,啶虫脒的平均添加回收率在76.4%～89.8%之间,相对标准偏差在1.3%～8.3%之间,啶虫脒的检出限为0.0075μg/mL。结论该方法操作简便,结果准确,适用于人血中啶虫脒的检验。     

气质联用SIM技术在毒物检测中的应用

目的　介绍气质联用SIM技术在毒物检测中的应用方法。方法　通过两个案例对质谱的两种扫描方法fullscan(全扫描 )和SIM (选择离子检验 )进行了比较。结果　SIM技术检验灵敏度远远高于Full Scan技术。结论　SIM技术是最理想的微量毒物检测技术之一。     

尿中苯丙胺、甲基苯丙胺、MDA和MDMA的固相微萃取和GC/MS/SIM测定

目的研究固相微萃取(SPME)用于尿中苯丙胺(AMP)、甲基苯丙胺(MET)、3,4-亚甲二氧基苯丙胺(MDA)和3,4-亚甲二氧基甲基苯丙胺(MDMA)的提取。方法样品调节至碱性和用盐饱和后用顶空SPME,内标为MET-d5。萃取纤维为100μm聚二甲基硅氧烷(PDMS)。用气质联用选择离子检测(GC/MS/SIM)。结果0.2μg/ml加标尿样,AMP、MET、MDA和MDMA的富集倍数分别为22,60,13和47。检出限(S/N=3)为0.4～9.5ng/ml。线性范围为0.05～1μg/ml。0.2、0.5和1.0μg/ml加标尿样,相对回收率77.9%～112.4%,变异系数2.7%～18.0%(n=5)。用该方法分析5个案件样品,和常规液液萃取结果接近。结论顶空SPME法用于尿中AMP、MET、MDA和MDMA等化合物的分析,无需有机溶剂,富集效率高,提取-富集-进样一体化,简单方便实用。     

A New Approach for the Carbon Monoxide (CO) Exposure Diagnosis: Measurement of Total CO in Human Blood Versus Carboxyhemoglobin (HbCO)

The aim of the study is to present the application of a headspace–gas chromatography–mass spectrometry (HS‐GC‐MS) method for the determination of the carbon monoxide (CO) blood concentration and to compare it with carboxyhemoglobin (HbCO) saturation. In postmortem cases, the HbCO measured by spectrophotometry frequently leads to inaccurate results due to inadequate samples or analyses. The true role of CO intoxication in the death of a person could be misclassified. The estimation of HbCO from HS‐GC‐MS CO measurements provides helpful information by determining the total CO levels (CO linked to hemoglobin (HbCO) and CO dissociated from hemoglobin). The CO concentrations were converted in HbCO saturation levels to define cutoff blood CO values. CO limits were defined as less than 1 μmol/mL for living persons, less than 1.5 μmol/mL for dead persons without CO exposure, and greater than 3 μmol/mL for dead persons with clear CO poisoning.     

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.     

Screening and detection of amphetamine derivatives in biological stains

A GC/MS-method is described for the screening, detection and determination of the commonly used drugs amphetamine, methamphetamine, MDA, MDMA and MBDB in small blood samples and bloodstains using solid phase SPE columns and a pipetting robot (Gilson Aspec XL). The detection limits are in the order of 0.03 to 0.08 microgram/L and the correlation factors between 0.9982 and 0.9998. Furthermore the stability was investigated covering a storage time of 64 days. The method has proven useful in forensic cases with only small sample volumes or bloodstains.     

Simple and simultaneous analysis of fenfluramine, amphetamine and methamphetamine in whole blood by gas chromatography-mass spectrometry after headspace-solid phase microextraction and derivatization

A simple and sensitive method for the simultaneous analysis of fenfluramine, amphetamine and methamphetamine in whole blood was developed using a headspace-solid phase microextraction (SPME) and derivatization. A 0.5 g whole blood sample, 5 microl d(5)-methamphetamine (50 micrig/ml) as an internal standard, and 0.5 ml sodium hydroxide (1 M) were placed into a 12 ml vial, and sealed rapidly with a silicone septum and an aluminum cap. Immediately after the vial was heated to 70 degrees C in an aluminium block heater, the needle of the SPME device was inserted through the septum of the vial, and the extraction fiber was exposed in the headspace for 15 min. First, heptafluorobutyric anhydride was injected into the injection port of the GC-MS, and the compounds extracted by the fiber were then desorbed and derivatized simultaneously by exposing the fiber in the injection port. The calibration curves, using an internal standard method, demonstrated good linearity throughout the concentration range from 0.01 to 1.0 microg/g. The detection limits of this method were 5.0 ng/g for fenfluramine and methamphetamine, and 10 ng/g for amphetamine. No interferences were found, and the time for analysis was about 30 min for one sample. This method was applied to a suicide case in which the victim ingested fenfluramine. Fenfluramine was detected in the blood sample collected from the victim at the concentration of 7.7 microg/g.