气相色谱/燃烧炉/同位素比值质谱方法对尿中睾酮来源的检测

目的 :建立检测尿中睾酮来源的方法 ,以确证睾酮阳性。方法 :采用薄层层析方法对尿样中内源性类固醇激素进行分离 ,再经气相色谱 /燃烧炉 /同位素比值质谱方法检测睾酮及其它内源性类固醇激素的δ值。结果 :薄层层析方法可以对尿样中内源性类固醇激素进行初步分离。样品中内源性类固醇激素δ值的比值为An ./PT:0 97;Etio./PT:1 0 7;T/PT :1 3 0 ;5α -diol/PT :1 1 1。结论 :利用此方法可以检测尿中睾酮的来源     

同位素比质谱法检测尿中类固醇来源

目的:采用同位素比质谱方法对使用某些类固醇制剂(本文选用雄烯二酮)后的尿样中的代谢物进行同位素比值测定,以检测尿中类固醇的来源,区分是自然代谢物还是使用类固醇制剂后的代谢物,并确定使用制剂后用此方法检测到阳性结果的时间。方法:采用酶解、高效液相色谱(HPLC)等方法对尿样中内源性类固醇激素进行酶解游离型、分离,再经气相色谱/燃烧炉/同位素比值质谱方法检测代谢物及内源性类固醇参照物13C与12C同位素含量的相对比值(δ值)。将被检测物与参照物同位素比值的差值与判别标准值比较,进行来源判定,并用正常人尿样进行对照。结果:使用雄烯二酮制剂后尿样中内源性类固醇激素代谢物δ值低于未使用的正常值,提示有外源性类固醇摄入。用同位素比质谱方法可在4或5天之内检测到阳性结果。结论:利用同位素比质谱法可在一定时间内检测尿中类固醇代谢物的来源。     

运动员尿中类固醇激素水平的研究

目的:对1217名运动员尿中8种类固醇激素的含量进行测定及统计分析,以考察我国运动员内源性类固醇激素水平.方法:对尿样进行预提取、水解、萃取、衍生化等前处理,采用GC/MS方法分离检测,测定内源性类固醇激素含量.结果:我国男运动员尿中类固醇水平一般高于女性,但都普遍低于国外统计数据,尤其是睾酮和5β雄烷二醇的水平低于国外数据4～7倍.某些类固醇含量的比值(如An/Etio、T/ET、5α-diol/5β-diol)较为恒定,且受性别因素影响较小.     

高效液相色谱/质谱法检测人尿中的糖皮质激素及其它兴奋剂

采用液相色谱/质谱(LC/MS)方法检测糖皮质激素、甾体和β2激动剂。本文采用Zorbax 30 mm×2.1mm,1.8μm短柱,对12种糖皮质激素、3种甾体和2种β2激动剂的液相色谱/质谱的测定方法进行了研究,建立了6分钟内快速检测人尿中多种糖皮质激素、甾体和β2激动剂的测定方法。检测限低于5ng/ml。该方法适用于兴奋剂的筛选及确证。     

营养补充品中刺激剂的分析测定

介绍营养补充品中刺激剂的毛细管气相色谱和气相色谱质谱联用检测方法.试样采用碱提,液-液分配提取,以HP-5毛细管柱为分离柱,用氮磷检测器测定.实验结果表明:该方法操作简便,分析速度快,结果可靠.添加平均回收率为75.4%～143.1%,相对标准偏差为0.27%～5.5%,检出限为5～40μg/L.     

氨溴特罗片在健康人体的药代动力学研究

目的研究单剂量及多剂量口服氨溴特罗片在中国健康成年志愿者人体药代动力学。方法 9名健康成年受试者单次口服氨溴特罗片2片（含盐酸氨溴索60mg,盐酸克仑特罗40μg）,单次给药试验结束后进行多次口服给药,每日早晨给药2片,共给药9d,达稳态后进行多次给药临床药代动力学研究,采用液相色谱-串联质谱联用分别测定给药后氨溴索及克仑特罗的血药浓度,应用DASver2.0软件进行房室拟合及参数计算。结果血中氨溴索和克仑特罗的血药浓度-时间数据均符合二室药物动力学模型,氨溴索的单次与多次给药的药代动力学参数无统计学差异,而克仑特罗多次口服达稳态后的清除率（CL/F）和表观分布容积（V/F）降低,峰浓度（Cmax）、曲线下面积（AUC（0-t）、AUC（0-∞））、消除速率常数K10、平均滞留时间（MRT）均明显增加。男女受试者的主要药动学参数无显著性差异。试验期间未发现明显不良反应。结论受试者连续口服氨溴特罗片,氨溴索在体内无药物蓄积作用,而克仑特罗有明显药物蓄积。主要代谢参数在我国健康成人中无性别差异。所建立的检测方法灵敏、准确、可靠、特异性强,可满足氨溴索和克仑特罗临床药动学试验要求。     

兴奋剂19-去甲雄酮和19-去甲本胆烷醇酮的检测研究

目的:19-去甲雄酮(19-Norandrosterone,19NA)和19-去甲本胆烷醇酮(19-Noretiocholanolone,19NE)是竞技体育中禁用的内源性类固醇兴奋剂,可由诺龙代谢而来。本研究的目的是采用同位素比质谱方法对尿中19NA和19NE进行检测规律研究。方法:征集2名男性志愿者。口服20 mg诺龙,并于服药前后收集尿样。对尿样中19NA和19NE的浓度及同位素比进行测定。结果:服药后尿样中19NA和19NE的浓度迅速上升,其同位素比降低至接近受试品的同位素比值。结论:摄入诺龙不影响尿中孕烷二醇、雄酮、本胆烷醇酮的同位素比值。尿样在服药后27小时内提示尿样为阳性。     

液相色谱-质谱法同时测定大鼠血浆中游离型神经甾体

目的建立同时测定SD大鼠血浆中脱氢表雄酮(DHEA)、孕烯醇酮(PREG)和别孕烯醇酮(AP)的高效液相色谱质谱法。方法选择甲睾酮(MT)作为内标。雄性SD大鼠的血浆使用液液萃取后,再经固相萃取、衍生化后,以高效液相色谱质谱检测器测定DHEA、PREG和AP的浓度。采用大气压化学离子源(HPLC/APCIMS),选择离子检测方式。结果血浆中DHEA、PREG和AP线性范围分别为:0.030～2.00ng·ml-1,0.025～2.00ng·ml-1,相关系数分别为0.9988、0.9996和0.9988。低、中、高浓度血样的提取回收率在70.4%～92.4%之间,相对回收率为:92.6%～105.6%,日内、日间RSD均小于15%。正常雄性SD大鼠的血浆PREG和AP分别为0.29±0.24ng·ml-1、0.19±0.20ng·ml-1,每ml血浆DHEA含量均低于定量限。结论本实验方法具有灵敏度高、准确度好及变异较小的特点,适合测定雄性SD大鼠血浆中3种游离型神经甾体的含量。     

普卢利沙星片在健康人体的药代动力学研究

目的研究单剂量口服普卢利沙星片在中国健康志愿者体内的药代动力学。方法12名健康成年受试者按3×3拉丁方随机分组,分别单次口服100、200、400mg普卢利沙星片,采用LC/MS/MS法测定给药后普卢利沙星代谢产物NM394的血药浓度,应用DAS ver2.0软件进行房室拟合及参数计算。结果血样中未检测到普卢利沙星,只能测定其代谢产物NM394。低、中、高3个剂量组均符合二室模型,在人体内的药动学过程符合一级动力学。男女受试者的主要药动学参数无显著性差异。试验期间未发现明显不良反应。结论普卢利沙星片单次3剂量口服给药后,其代谢产物NM394的Cmax和AUC呈良好的剂量依赖性,主要代谢参数在我国健康成人中无性别差异。检测方法灵敏、准确、可靠、特异性强,可满足普卢利沙星临床药动学试验要求。     

固相萃取-气相色谱-串联质谱法检测粮食作物中的T-2与HT-2毒素

目的建立粮食作物中T-2与HT-2毒素的高灵敏度气相色谱-串联质谱(GC-MS/MS)检测方法。方法染毒的粮食样品以柱淋洗方式用甲醇提取后,采用C18固相萃取柱净化富集,再经七氟丁酰咪唑衍生化后,在气相色谱-串联质谱仪上采用选择反应监测(SRM)模式进行检测。结果在染毒大米、小米、糯米和玉米样品中,T-2与HT-2毒素在0.5～100μg/kg范围内均呈现良好的线性关系。在最优分析条件下,两种毒素的最低检出限均在0.05～0.5μg/kg。在2、30、60μg/kg 3个加标浓度下,T-2与HT-2毒素的方法回收率均在47.2%～110.8%。结论该方法操作简便,灵敏度高,适用于各类粮食样品中T-2与HT-2毒素的检测。     

Specific interpretation of hair concentrations in 2 fatal metformin intoxication cases

Hair analysis is very useful for toxicological investigations since, by providing a wider detection window, it gives the possibility to perform a retrospective study on the historical consumption of a substance. Unfortunately, there are no data available for hair concentrations in metformin-related deaths. In this study, the authors present 2 cases of fatal metformin intoxication in which, for the first time, hair analysis was performed using a specific GC–MS/MS method. Metformin was tested positive in femoral blood (112.3 mg/L and 64.7 mg/L respectively) and cardiac blood (226.9 and 203.2 mg/L) of the two subjects. For case 1, other samples were also tested positive, including vitreous humor (31.1 mg/L) and gastric contents (773.5 mg/L). In case 2, metformin was measured at 844.9 mg/L in urine. Metformin hair concentrations were 28.3–44.8 and 22.5 ng/mg for both cases, respectively. The concentrations found in the 2 fatal cases are clearly higher than those obtained in a previous study with subjects under treatment (0.3–3.8 ng/mg) or those found in 3 post-mortem cases where metformin death was excluded (0.6–1.4 ng/mg). Excessive sweating during the agonal phase due to fatal hypoglycemia could explain these elevated concentrations as sweat can have contaminated the hair.     

Dose-concentration relationships of methadone and EDDP in hair of patients on a methadone-maintenance program

After controlled oral administration of d,l-methadone solution (15–260 mg/day) in the context of a methadone-maintenance program, concentrations of methadone and 2-ethylidine-1,5-dimethyl-3,3-diphenyl-l-pyrrolidine (EDDP), in head hair were determined (N=41), using a fully automated headspace solid-phase microextraction procedure in combination with gas chromatography and mass spectrometry (HS-SPME/GC/MS). Methadone was present in all samples in concentrations ranging from 0.25 to 13.29 ng/mg (mean 2.69±0.45 ng/mg). EDDP was also present in every sample in concentrations ranging from 0.05 to 2.17 ng/mg (mean 0.43±0.08). The concentration ratio methadone/EDDP was 7.5±5.7 in the proximal segments, but decreased to 4.8±1.4 in the distal segments. A statistically significant correlation between the intake dose and the methadone and EDDP concentrations in the subjects’ hair could be established only in the proximal segments (r=0.913 for methadone and r=0.901 for EDDP), but not in the distal segments. In all, 131 segments analyzed, the correlation coefficient was r=0.760 for methadone and r=0.738 for EDDP. In comparison to the dose-concentration relationship reported in the literature, we found a better correlation with higher correlation coefficients especially in the proximal segments. However, owing to a broad distribution in the correlation between dosage and concentration, the determination of methadone and EDDP in hair holds only limited information about prior methadone administration.     

Detection of imidacloprid in biological fluids in a case of fatal insecticide intoxication

Here, we describe a high-performance liquid chromatography/photodiode array detector method for the detection of imidacloprid in biological fluids in a case of suicide by ingestion of liquor mixed with Admire® Flowable insecticide (containing 20% imidacloprid). A plastic bottle containing a cloudy liquid (concentration of ethanol in the liquid was 150 mg/ml and that of imidacloprid was 50 mg/ml) was found near the decedent. The biological fluids collected at autopsy were prepared by deproteinization with acetonitrile. Zolpidem was used as an internal standard. The concentrations of imidacloprid in femoral blood and cerebrospinal fluid were 105 and 58.5 μg/ml, respectively. Ethanol was also detected in the samples, with concentrations of 1.0 mg/ml in femoral blood and 1.4 mg/ml in cerebrospinal fluid.     

Toxicological analysis of drugs and poisons in formalin-fixed organ tissues

Diethylether, chloroform and toluene were administered by inhalation and ethanol intravenously to rabbits. As soon as possible after death, tissue specimens were collected from the brain, lung, liver, kidney and skeletal muscle and fixed in non-buffered 10% formalin at room temperature (10–20° C) for 4 different periods (1, 2, 5 and 14 days). The volatile substances were analyzed and identified by gas chromatography/mass spectrometry (GC-MS). The measured concentrations of ethanol, diethylether, chloroform and toluene in the brain tissue 1 day after fixation decreased to 8, 23, 73 and 84% respectively compared with those in the non-fixed brain tissue (100%). The rank order of the rate of decrease in the fixed state was: ethanol > diethylether chloroform > toluene. These volatile substances could be detected clearly in all the tissue specimens, even after a 14-day fixation period. These results provide useful toxicological information that will help to differentiate whether volatile substances have been administered antemortem or postmortem.     

硒缺乏和训练对雄性大鼠血清睾酮的影响

通过建立硒缺乏运动模型,结合游泳训练,观察硒缺乏和运动训练对雄性大鼠血清睾酮的影响。结果显示：①用低硒饲料喂养７周的大鼠的血清硒含量明显低于用补硒饲料喂养的大鼠。②缺硒安静大鼠的睾丸硒含量低于补硒安静大鼠,但降低的幅度比血清硒降低的幅度小,训练使缺硒大鼠的睾丸硒含量明显升高。③硒缺乏和训练均使大鼠的血清睾酮和游离睾酮水平降低,这种降低与血清的促性腺激素（ｒＬＨ和ｒＦＳＨ）水平无关,与睾丸Ｌｅｙｄｉｇ细胞膜上的ＬＨ／ｈＣＧ受体的亲和性及结合容量无关。④硒缺乏和训练均不影响血清的皮质酮水平,睾酮／皮质酮比值和游离睾酮／皮质酮比值的降低,主要是因为睾酮和游离睾酮的降低而引起的     

GC／MS法测定人血浆中地芬尼多药物浓度及其在药代动力学研究中的应用

目的建立GC/MS法测定人血浆中地芬尼多的含量,用于药代动力学研究中血药浓度的测定。方法液-液萃取法进行血样前处理,分析采用（15 m×0.25 mm×0.25μm）DB-17MS毛细管柱,载气为氦气,载气流速为1.0 ml/m in;进样口温度250℃,炉温升程：100℃→280℃（按25℃/m in的速度升温,且在280℃保持1 min）再升至300℃（按50℃/min的速度升温,且在300℃保持1.6 m in）;MS源温度230℃,四级杆温度150℃,电子轰击能量为70 eV。结果地芬尼多及内标的保留时间分别为7.8 min及7.97 min,其定量碎片基峰分别为m/z98和138。地芬尼多在1300 ng/ml浓度范围内,浓度与响应呈良好的线性关系（r2=0.999）;定量下限为1 ng/ml,准确度在82%104%范围内;地芬尼多的提取回收率为72.7%90.0%;批内和批间RSD分别在2.7%和3.9%以内,准确度在101.5%114.1%范围内;地芬尼多人血浆样品在-20℃保存10 d及在室温放置24 h稳定;其甲醇溶液在4℃冰箱保存21 d稳定。结论本研究建立的GC/MS方法测定人血浆中地芬尼多浓度,特异性好、灵敏度高,适用于盐酸地芬尼多口崩片在健康受试者体内的药代动力学研究。     

Effects of blood sampling technique and exercise on plasma androgens in female rats.

Plasma concentrations of testosterone, androstenedione, and dihydrotestosterone rise during exercise in women. The purpose of this study was to evaluate exercise-induced changes in these three androgens in female rats to see if the rat may be used as a model for studying these responses. Because some androgen originates in the adrenal cortex, and the adrenocortical axis is stress-responsive, blood samples taken by cardiac puncture under halothane anesthesia, a potentially stressful technique, were compared with those taken by decapitation, which should not elicit a stress response. Blood was collected from sedentary rats at rest and from exercise-trained rats after 30 min of running. Half the samples were taken by each technique. Sampling technique did not affect androgen levels. Combining data from both techniques, testosterone was similar in rats at rest (503 pmol.l-1 +/- 62 (SE), N = 20) and post-exercise (500 +/- 31, N = 17). Androstenedione was higher post-exercise (1648 pmol.l-1 +/- 248, N = 17) than at rest (866 +/- 115, N = 18; P less than 0.009). Dihydrotestosterone was also higher post-exercise (452 pmol.l-1 +/- 31, N = 17) than at rest (324 +/- 41, N = 20; P less than 0.05). It is concluded that: 1) cardiac puncture with halothane anesthesia is acceptable for studying androgens in female rats, and 2) the female rat may be useful for studying the androstenedione and dihydrotestosterone responses to exercise, but is not appropriate for studying testosterone exercise responses.     

禁用药物β_2-激动剂的分析研究

采用灵敏度很高的气相色谱 /负离子化学电离质谱 (GC/NICIMS)对 9种国际奥委会禁用药物 β2 -激动剂进行分析研究 ,比较了不同衍生化方法得到的产物在NICIMS上的响应。确定了五氟丙酸酐衍生化产物有最佳的灵敏度 ,适合此类药物的检测要求。重点对克伦特罗尿样进行了检测 ,其结果符合实际工作的需要     

Determination of dihydrocodeine in hair of opiate addicts by GC/MS

Summary After the examination of more than 300 hair samples of suspected heroin abusers, a large number of which proved positive, we can say that high concentrations of dihydrocodeine can be determined either in addition to, or in the place of, morphine and also frequently in combination with codeine. The opiates were extracted after dissolving the hair samples in NaOH and hydrolysis with HCI. The quantitative determination of dihydrocodeine was achieved by derivatisation with HFBA using GC/MS at m/u = 497. Dihydrocodeine is used in antitussive drugs. After the examination of individual hair samples, it was obvious that some heroin consumers had switched to dihydrocodeine. This may lead to the conclusion that dihydrocodeine itself is used either as an intoxicating drug or to reduce withdrawal symptoms. The increasing number of positive samples should be noted by the legal authorities.     

A fatal poisoning case of acetone cyanohydrin and citalopram

Acetone cyanohydrin (ACH) is a readily available source of cyanide and is widely used in basic and applied sciences. In toxicology, ACH is classified as extremely hazardous as it readily decomposes on contact with water, with the potential rapid release of highly toxic hydrogen cyanide (HCN). We report the case of a young woman found dead from the intentional ingestion of ACH and citalopram, an antidepressant of the selective serotonin reuptake inhibitor class. The autopsy findings included bright reddish-purple hypostasis and mild pulmonary edema. As ACH can decompose to acetone and HCN, we quantified the concentration of each compound and thiocyanate separately in various body fluids and organs and determined their whole-body distributions by using gas chromatography–mass spectrometry (GC–MS). We observed high concentrations of both acetone and cyanide in the blood (0.63 mg/mL and 17.99 mM, respectively) and gastric contents (9.76 mg/mL and 472.44 mM). The whole-body distributions of acetone and cyanide were similar (i.e., the concentration of each compound was the highest in the lung, followed by the heart, and then the liver). Our results suggest that not only the route of administration but also the dose taken could greatly affect the body distributions of cyanide in humans. In addition, as toxicological screening detected citalopram, which was not prescribed to the deceased, we performed a chiral analysis by using liquid chromatography–tandem mass spectrometry (LC–MS/MS). We determined that only (S)-citalopram was ingested antemortem; its concentration was 0.36 μg/mL, which is in the toxic range.