阿片类成瘾者血清、尿中吗啡TLCS分析

建立阿片类成瘾者血清、尿液中吗啡的薄层色谱扫描 (TCLS)定量检测方法。样品经酸、碱性水解后调至pH9,氯仿 /异丙醇 ( 9∶1)萃取及GDX 40 3柱固相萃取 ,在紫外区可见光区薄层扫描。测得 3种萃取方法吗啡回收率分别为 75 3 %± 4 9% ,80 9%± 3 2 %和 79 4%± 3 5 % ,血清、尿中吗啡最低检出浓度分别为 0 1μg·ml-1,0 0 5 μg·ml-1(信噪比≥ 3 )。本法可用于阿片类药物成瘾者或中毒者血、尿中吗啡的检测。     

尿中吗啡的氮磷检测——气相色谱分析法

目的建立尿中吗啡的简便快速、灵敏可靠的GC/NPD分析方法。方法样品尿加内标烯丙吗啡,酶或酸催化水解,氯仿-异丙醇(9:1)液液提取或GDX403树脂固相提取,BSA衍生化,HP-5柱和氮磷检测器进行分析。结果 提取率62％～85％,检出限1.2～3.1ng/ml,线性范围20～2000ng/ml,回收率(97％～99％)±(6％~9％)(Mean±cv,N=5)。结论 方法适合于实际案件中尿样的检验。     

Detection and quantification of low levels of benzoylecgonine in equine urine

Cocaine (COC) is a highly addictive plant alkaloid expressing strong psychostimulatory effect. It has no medical use in equine veterinary practice. The contamination of the environment with cocaine such as its presence on the US paper currency has been reported few times. There are anecdotal reports of low benzoylecgonine (BE) concentrations (usually much less than 100 ng/mL) being found in urine of race horses. In order to protect horsemen against harsh penalties associated with the presence of trace amounts of BE in horse urine as a result of environmental contamination, in February 2005 the Illinois Racing Board issued new medication rules that established the threshold level of 150 ng/mL for BE in equine urine. The penalties associated with this rule provide for increasing fines ($250, $500, $1000) with successive positive reports against a trainer for levels of BE below 150 ng/mL. A total of 19,315 urine samples were collected over the 2-year period of time from winning horses (both harness and thoroughbred) at race tracks in Illinois for routine drug screening (ELISA). The presence of BE was confirmed by GC/MS in 28 urine samples (0.14%). The concentration range for BE in harness horses (21 detections) was < 5-91 ng/mL, and for thoroughbred (seven detections) was 7-52 ng/mL. To date, the laboratory has not reported concentrations of BE that exceed the established threshold concentration of 150 ng/mL.     

Gas chromatographic/mass spectrometric analysis of morphine and codeine in human urine of poppy seed eaters

In this study, poppy seeds were examined for a natural constituent that might serve as a maker for the seeds' ingestion as opposed to opiate abuse. Thebaine was selected as possible marker, since it was found to be a component of all poppy seeds examined and was not a natural component of different heroin samples. During the course of this investigation, a new extraction and cleanup procedure was developed for the gas chromatographic/nitrogen phosphorus detection (GC/NPD) and gas chromatographic/mass spectrometric (GC/MS) analysis of morphine and codeine in urine. A linear response, over a concentration range of 25 to 600 ng/mL, was obtained for codeine and morphine (r = 0.9982 and 0.9947, respectively). The minimum detectable level (LOD) and limit of quantitation (LOQ) for morphine were 10 and 30 ng/mL, respectively; whereas LOD and LOQ for codeine were 2 and 8 ng/mL, respectively. The coefficients of variance (CV, n = 6) for morphine and codeine analyses at the 100-ng/mL level were 13.3 and 4.6%, respectively. This procedure was used for the analysis of urine samples from five poppy seed eaters who each ingested 200 g of poppy seed cake. Results indicated that significant amounts of morphine and codeine are excreted in urine and that in all subjects, at least at one point in time, the apparent morphine concentration as determined by radioimmunoassay (RIA) analysis exceeded the cutoff value (300 ng/mL) established for screening. Thebaine was not detected in urine specimens collected following poppy seeds ingestion and thus could not be used as a marker.     

Opiate levels in hair

By means of radioimmunoassay-technique, hair samples of users, drug related fatalities, carcinoma patients receiving morphine and of experimental guinea pigs receiving codeine were investigated for opiates. The RIA-investigations require a minimum of material; our routine procedures need only 50 mg of hair. No correlation existed between administered doses of opiates and their concentrations in hair of both human and experimental animals. By sectioning the hair, the approximate period of drug use in man could be detected. However, these findings could not be confirmed by the animal experiments. The growth rate of the hair, diffusion and adhesion processes may influence the transport of drugs along the hair. External contaminations and washing procedures were shown to increase or diminish the drug concentration of the samples.     

Application of solvent microextraction to the analysis of amphetamines and phencyclidine in urine

A fast and simple method to detect some commonly abused illicit drugs, amphetamine, methamphetamine, 3,4-methylendioxy-amphetamine (MDA), 3,4-methylendioxy-methamphetamine (MDMA), 3,4-methylendioxy-N-ethylamphetamine (MDEA) and phencyclidine (PCP) in urine using solvent microextraction (SME) combined with gas chromatography (GC) analysis has been developed. The extraction is conducted by suspending a 2 μl drop of chloroform in a 2 ml urine sample. Following 8 min of extraction, the organic solvent is withdrawn into the syringe and injected into a GC with a pulsed discharge helium ionization detector (PDHID).The effects of different extraction solvents and times, pH and sample preparation were studied. The optimized method was capable of detecting drugs in urine at concentrations below Substance Abuse and Mental Health Services Administration (SAMHSA) established cut-off values for preliminary testing. Good linearity and reproducibility of extraction were obtained. The limits of detection were 0.5 μg/ml for amphetamine, 0.1 μg/ml for methamphetamine and MDA, 0.05 μg/ml for MDMA, 0.025 μg/ml for MDEA and 0.015 μg/ml for PCP. Relative standard deviation (R.S.D.) values ranged between 5 and 20% for the studied drugs.     

The unreliability of using a urine ethanol concentration to predict a blood ethanol concentration

Of approximately 5,000 forensic cases with a positive ethanol result, over 1,000 were available in which both blood and urine were present for comparison of ethanol content. Data were examined for calculation of the urine to blood ethanol concentration ratio, with the intent of evaluating the validity of predicting a blood ethanol level given a urine ethanol level. The overall urine to blood ethanol concentration ratio was 1.57:1 with a range of 0.7 to 21.0:1. The extremely wide range of values implies that a large degree of error would be introduced if a mean ratio was used when predicting a blood ethanol level from a urine ethanol level.     

内标法固相提取-GC/MS检测海洛因吸食者尿液中代谢物

本文建立了以SKF-525A作内标、GDX－301作固相提取剂、用GC／MS方法测定海洛因吸食者尿液中代谢物浓度的方法。系统考查了吗啡、单乙酰吗啡的提取回收率、线性关系等指标。所建立的方法对吗啡和单乙酰吗啡的回收率均大于80％;最小检测浓度小于0.1μg／ml;用代谢物特征离子与内标物特征离子峰高比法测定吗啡与单乙酰吗啡的浓度,在每ml尿添加0．25μg～250μg范围内,二者均有良好的提取线性关系。并对几例海洛因滥用者的尿液进行了检测。     

Roadside detection of impairment under the influence of ketamine--evaluation of ketamine impairment symptoms with reference to its concentration in oral fluid and urine

Although there are many roadside testing devices available for the screening of abused drugs, none of them can be used for the detection of ketamine, a popular abused drug in Hong Kong. In connection to local drug driving legislation, effective roadside detection of ketamine in suspected drug-impaired drivers has to be established. According to the drug evaluation and classification program (DEC), ketamine is classified in the phencyclidine (PCP) category. However, no study has been performed regarding the signs and symptoms exhibited by users under the influence of ketamine. In a study to develop a protocol for effective roadside detection of drug-impaired drivers, 62 volunteers exiting from discos were assessed using field impairment tests (FIT) that included measurements of three vital signs (i.e. body temperature, pulse rate and blood pressure), three eye examinations [pupil size, lack of convergence (LOC) and horizontal gaze nystagmus (HGN)] and four divided attention tests (Romberg, one-leg stand, finger-to-nose and walk-and-turn tests). Subsequent laboratory analysis of oral fluid and urine samples from the participants revealed the presence of common abused drugs in both the urine and oral fluid samples of 55 subjects. The remaining 7 subjects with no drug in their oral fluid samples were used as drug-free subjects. In addition, 10 volunteers from the laboratory who were regarded as drug-free subjects were also assessed using the same FIT. Among the 62 volunteers, 39 of them were detected with ketamine in their oral fluid. Of these ketamine users, 21 of them (54%) with only ketamine found in their oral fluid samples while the rest (18 subjects) of them had other drugs (i.e. MA, MDMA, benzodiazepines and/or THC) in addition to ketamine. Of the 21 ketamine-only users, 15 of them (71%) were successfully identified by FIT. It was found that when salivary ketamine concentrations were greater than 300 ng/mL, signs of impairment became evident, with over 90% detection rate using the FIT. By comparing the FIT observations on the 21 ketamine-only users with the drug-free subjects, the typical signs and symptoms observable for subjects under the influence of ketamine included LOC, HGN, elevated pulse rate and in general, failing the divided attention tests, especially the walk-and-turn and one-leg stand.     

尿中地西泮及其代谢物的气相色谱电子捕获检测法分析

目的　建立尿中地西泮及其代谢物替马西泮、去甲西泮和奥沙西泮的GC/ECD分析方法。方法　尿样经β 葡萄糖醛酸酶水解后 ,用有机溶剂提取 ,再以N ,O 双三甲硅烷基三氟乙酰胺进行衍生化 ,衍生物用GC/ECD法分析。结果　检材中分析物的回收率 70 %以上 ,检出限 5ng/ml以下。结论　本法可进行口服地西泮 10mg人体 48h内尿液中地西泮代谢物的分析。     

固相萃取同时提取尿中的39种药毒物

目的建立固相萃取方法同时提取尿中的39种药毒物并用高效液相色谱法进行分析。方法以多沙普仑为内标,1ml尿样经Oasis小柱固相萃取,用HPLC进行分析。结果39种药毒物可同时从尿中提出,内源性物质不干扰测定。其绝对回收率除吗啡外均大于70％;天内及天间精密度均小于10％;检测限1-15ng／ml;线性相关系数在0.9977以上。结论本法快速、简便、重现性好,空白干扰小,可用于实际案例的药物毒物筛选。     

A high selective screening test for methamphetamine in human urine

A high selective screening test (Ad-Tip method) for methamphetamine in human urine has been devised. The method involves a brief extraction from a urine sample with an Ad-Tip (ODS-silica minicolumn), washing, eluting with modified Simon's reagent, and coloration with carbonate buffer. Detection limit of methamphetamine in urine is 1 microgram/ml and the test takes within 3 min/sample. The results of the Ad-Tip method were almost identical to those of laboratory analyses.     

高效薄层色谱法检测尿中硝西泮的代谢物7-氨基硝西泮

目的 建立用高效薄层色谱法定性及半定量测定人尿中硝西泮的代谢物7-氨基硝西泮(7ANIZ)含量的方法。方法 人口服治疗量10 mg硝西泮后,在pH 9条件下用乙醚进行提取,分析物斑点用氟罗里丝进行荧光显色,紫外灯下(366nm)观察荧光斑点;根据斑点荧光显色情况及强度进行7ANIZ定性及半定量检测。结果 尿中硝西泮代谢物7ANIZ检出限为5 ng/ml,测量限为15 ng/ml。结论 人口服治疗量10 mg硝西泮,用高效薄层色谱法可定性及半定量测定48 h内排泄尿中的7ANIZ。     

Morphine to codeine concentration ratio in blood and urine as a marker of illicit heroin use in forensic autopsy samples

A morphine to codeine ratio greater than unity (M/C>1) has been suggested as an indicator of heroin use in living individuals. The aim of this study was to examine the morphine to codeine ratio in a large population (N=2438) of forensically examined autopsy cases positive for 6-monoacetylmorphine (6-MAM) and/or morphine in blood and/or urine. Blood and urine concentrations of 6-MAM, morphine and codeine were examined using GC-MS and LC-MS/MS methods. In 6-MAM positive samples, the M/C ratio was greater than unity in 98% (N=917) of the blood samples and 96% (N=665) of the urine samples. Stratification of 6-MAM negative cases by M/C above or below unity revealed similarities in morphine and codeine concentrations in cases where M/C>1 and 6-MAM positive cases. Median blood and urine morphine concentrations were 8-10 times greater than codeine for both groups. Similarly to 6-MAM positive cases, 25-44 year-old men prevailed in the M/C>1 group. In comparison to cases where M/C ≤ 1, the M/C ratio was a hundred times higher in both 6-MAM positive and M/C>1 cases. The range of morphine concentration between the lowest and the highest quintile of codeine in M/C>1 cases was similar to that in 6-MAM positive cases. This range was much higher than for M/C ≤ 1 cases. Moreover, linear regression analyses, adjusted for age and gender, revealed a strong positive association between morphine and codeine in 6-MAM positive and M/C>1 cases. The M/C ratio appeared to be a good marker of heroin use in post-mortem cases. Both blood and urine M/C>1 can be used to separate heroin users from other cases positive for morphine and codeine.     

DPX-GC/MS法分析尿液中新精神活性物质二苯基乙基哌啶

目的 建立一种枪头式分散固相微萃取-气相色谱质谱法,分析尿液中的二苯基乙基哌啶.方法 对影响萃取效果的吸附树脂种类、淋洗液种类、洗脱液种类和洗脱液用量进行优化考察.结果 在最佳实验条件下,尿液中二苯基乙基哌啶的线性范围为0.02～1.0 mg/L,相关系数r2>0.99,检出限为0.005mg/L,回收率在76.7％～...     

多虑平SPE-HPLC分析方法的建立及其应用

目的　建立尿样和全血中多虑平的固相萃取 高效液相色谱 (SPE HPLC)分析方法。方法　以多沙普仑为内标 ,1ml尿样或 0 5ml全血用Oasis小柱固相萃取后进Lichrospher 10 0RP 18e ( 2 5 0mm× 4mm ,5 μm)分析柱进行分析 ,2 3 0、 2 5 0nm同时进行检测。结果　尿样和全血中的检测限均 2ng/ml,线性相关系数r≥ 0 9992 ,天内和天间精密度均小于 6 75 % ,绝对回收率大于 85 % ,内源性物质不干扰测定。结论　本法快速、简便、准确 ,可用于实际案例的检测。     

Simultaneous analysis of six amphetamines and analogues in hair, blood and urine by LC-ESI-MS/MS: Application to the determination of MDMA after low Ecstasy intake

A rapid and sensitive method using LC-MS/MS triple stage quadrupole for the determination of traces of amphetamine (AP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, “ecstasy”), 3,4-methylenedioxyethamphetamine (MDEA), and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB) in hair, blood and urine has been developed and validated. Chromatography was carried out on an Uptisphere ODB C₁₈ 5 μm, 2.1 mm × 150 mm column (Interchim, France) with a gradient of acetonitrile and formate 2 mM pH 3.0 buffer. Urine and blood were extracted with Toxitube A^® (Varian, France). Segmented scalp hair was treated by incubation 15 min at 80 °C in NaOH 1 M before liquid–liquid extraction with hexane/ethyl acetate (2/1, v/v). The limits of quantification (LOQ) in blood and urine were at 0.1 ng/mL for all analytes. In hair, LOQ was <5 pg/mg for MA, MDMA, MDEA and MBDB, at 14.7 pg/mg for AP and 15.7 pg/mg for MDA. Calibration curves were linear in the range 0.1–50 ng/mL in blood and urine; in the range 5–500 pg/mg for MA, MDMA, MDEA and MBDB, and 20–500 pg/mg for AP and MDA. Inter-day precisions were <13% for all analytes in all matrices. Accuracy was <20% in blood and urine at 1 and 50 ng/mL and <10% in hair at 20 and 250 pg/mg. This method was applied to the determination of MDMA in a forensic case of single administration of ecstasy to a 16-year-old female without her knowledge during a party. She suffered from hyperactivity, sweating and agitation. A first sample of urine was collected a few hours after (T + 12 h) and tested positive to amphetamines by immunoassay by a clinical laboratory. Blood and urine were sampled for forensic purposes at day 8 (D + 8) and scalp hair at day 60 (D + 60). No MDMA was detected in blood, but urine and hair were tested positive, respectively at 0.42 ng/mL and at 22 pg/mg in hair only in the segment corresponding to the period of the offence, while no MDA was detectable. This method allows the detection of MDMA up to 8 days in urine after single intake.     

SPE-GC／MS法检测大鼠尿中尼美西泮及其代谢物

目的检测尼美西痒阳性大鼠尿液中的代谢物。方法2只Wistar大鼠分别喂食半粒尼美西泮药丸，收集24h内尿液，用B葡萄糖醛酸酶水解，Oasis@HLB柱固相提取，以DB．35Ms柱为分离柱，气相色谱质谱联用法检测。结果从大鼠尿液中检出尼美西泮的4种代谢物：7-乙酰氨基尼美西泮、7-乙酰氨基硝基安定、7-氨基尼美西泮和2-氨基-5-硝基苯基苯甲酮及少量尼美西泮原体。结论尼美西泮在大鼠体内易代谢，在尿液中的主要代谢物为7-乙酰氨基尼美西泮和7-乙酰氨基硝基安定。     

人血、尿中碘解磷定的高效液相色谱分析

目的建立人血、尿中碘解磷定的高效液相色谱分析方法,用于法医学鉴定和指导临床合理用药。方法以空白人血浆和尿液分别添加标准碘解磷定对样品的前处理方法、仪器测试条件、定性、定量分析方法进行考察和优化。对疑似有机磷中毒抢救或死亡者血、尿中的碘解磷定浓度进行测定。结果血、尿中碘解磷定浓度的线性范围是0.5~8.0μg/m l;定量检测的浓度限为0.5μg/m l;日内、日间精密度RSD≤5.3%(n=5);回收率:血≥(96.7±2.9)%;尿≥(93.7±3.8)%。结论所建分析方法快速、准确,适应于法医学鉴定和临床救治中碘解磷定血、尿浓度监测等相关研究。     

Hair and urine testing to assess drugs of abuse consumption in couples undergoing assisted reproductive technology (ART)

For the first time in Europe hair and urine testing have been applied to assess drugs of abuse consumption in couples undergoing assisted reproductive technology and the eventual association of toxic habits with other lifestyle, health status and sociodemographic factors was also investigated. Couples attending five assisted reproduction centers in Rome were invited to join the study. When they presented at the Centre for the visit, they were asked to answer a structured questionnaire concerning sociodemographic characteristics and lifestyle habits, and at the same time to provide hair and urine samples. Hair and urine testing for drugs of abuse, urinary profile of principal endogenous steroids involved in fertility process (testosterone, epitestosterone, androsterone, etiocholanolone and dehydroepiandrosterone) and of alcohol and tobacco smoke biomarkers were performed with validated methodologies. Of the 594 enrolled individuals (297 couples), 352 (164 couples and 24 single individuals from the couple) completed the questionnaire and gave both hair and urine samples, apart from 3 bald men, who only gave urine samples. Urine testing showed an overall 4.8% (17 individuals) positivity to drugs of abuse: 4.2% to cannabinoids, 1.4% to cocaine and 0.85% to both drugs. Results of 4cm segment hair samples testing matched those from urine samples. Thus, taking together, results of urine and hair testing confirmed repeated use of cannabis, cocaine and both drugs in 3.7, 0.85 and 0.57% examined individuals, respectively. Drug consumers were in a statistically higher percentage active smokers and alcohol drinkers, less prone to physical activity and with a trend towards higher weight than non consumers. Finally, repeated drug consumption was associated with significant lower concentration of urinary testosterone in males and of urinary dehydroepiandrosterone in females. The findings of the present study confirm the suitability of urine testing to disclose recent drugs of abuse consumption and of hair analysis to verify repeated consumption. Association between different toxic habits and sedentary lifestyle is also substantiated by the obtained results in our cohort of couples attending assisted reproduction centers.