血液、尿液中氯胺酮及其代谢物去甲氯胺酮的HPLC分析

目的 建立血液、尿液中氯胺酮及其代谢物去甲氯胺酮的高效液相色谱(HPLC)分析方法.方法 以非那西丁为内标,检材加入10%的氢氧化钠溶液调节pH值为14,用甲苯提取,离心后取有机层,水浴下吹干,乙腈定容后进HPLC仪分析.结果 检测血液中氯胺酮和去甲氯胺酮的线性范围均是0.05～10μg/mL(r2＞0.999 3),检测尿液中氯胺酮和去甲氯胺酮的线性范围均是0.01～50 μg/mL(r2＞0.999 5).氯胺酮和去甲氯胺酮在血液和尿液中的检测限分别是0.006 μg/mL和0.003 μg/mL.血液和尿液中氯胺酮和去甲氯胺酮的回收率不低于82.4%.检测血液和尿液中氯胺酮和去甲氯胺酮的日内精密度和日间精密度均小于10.0%.将所建的方法应用于给大鼠氯胺酮后的血液和尿液中的氯胺酮和去甲氯胺酮的测定,得到了氯胺酮和去甲氯胺酮在大鼠的药时曲线和尿排药速率曲线. 结论本方法简便、快捷,适用于血液、尿液中氯胺酮及其代谢物去甲氯胺酮的分析.     

生物检材中氯胺酮及其代谢物的检测

近年来氯胺酮的滥用越来越普遍,建立快速、准确的检测方法越来越重要。氯胺酮在生物体内的代谢物主要有去甲氯胺酮、脱氢去甲氯胺酮等。目前,常用的生物检材有血液、尿液、毛发等。常用的检测方法有气相色谱法、气相色谱-质谱联用法、高效液相色谱法、液相色谱-质谱联用法、高效毛细管电泳法等。本文参考近年来的相关文献对生物检材中氯胺酮及其代谢物的检测方法作一综述,为法医毒物分析等相关领域提供参考。     

QTRAP LC-MS/MS方法分析头发中氟胺酮及其代谢物的研究

目的建立头发中氟胺酮及其代谢物的液相色谱-四极杆/线性离子阱质谱(QTRAP LC-MS/MS)检测方法并分析头发样本中氟胺酮含量范围。方法将洗净的20mg头发样本加入2mL提取液研磨后超声提取,离心取上清液过滤膜后,采用多反应监测模式测定氟胺酮及其代谢物,并以该方法分析了50例样本中氟胺酮的含量。基于氟胺酮、氯胺酮结构的相似性,参考氯胺酮代谢物去甲氯胺酮的质谱裂解途径,对氟胺酮主要代谢物进行推断。结果氟胺酮在浓度范围0.004ng/mg～2ng/mg内线性良好;方法检出限为0.001ng/mg;在0.05、0.20、1.00 ng/mg 3个添加水平的回收率为90.2%～94.4%。50例阳性样本中氟胺酮含量在0.2ng/mg以上46例占比92%,含量最高值92.56ng/mg、平均值15.32ng/mg、中位值5.34ng/mg,反映了氟胺酮较为严重的滥用形势。结论本方法简便、高效、可靠,适用于头发中氟胺酮及其代谢物的鉴定。样本数据为氟胺酮列管后鉴定及阈值确定提供了参考。     

家兔血液和尿液中氯胺酮浓度的相关性

目的研究家兔尿液中氯胺酮及代谢物去甲氯胺酮浓度与血药浓度的动态相关性。方法实验家兔分为氯胺酮灌胃组、静脉注射组和对照组,分别于染毒前和染毒后不同时间点收集尿液和血液。气相色谱/质谱联用(GC/MS)全扫描定性、气相色谱(GC)定量分析血液和尿液样品中氯胺酮及去甲氯胺酮的浓度。采用双变量Pearson相关分析研究尿液中药物浓度和血药浓度的相关性。结果氯胺酮灌胃组和静脉注射组给药后各时间点氯胺酮及去甲氯胺酮在尿液和血液中的浓度相关系数范围在0.11～0.69之间。结论氯胺酮及去甲氯胺酮在尿液和血液中的浓度相关性较差,尿液药物浓度并不能直接反映血药浓度,因此用尿液中氯胺酮浓度推断血药浓度时应慎重考虑。     

氯胺酮在家兔体内的毒物代谢动力学

目的研究氯胺酮及其代谢物去甲氯胺酮在家兔体内的毒物代谢动力学特征。方法家兔以氯胺酮0.15g/kg剂量灌胃,分别于给药前和给药后不同时间点收集血液和尿液,血清和尿液中氯胺酮及代谢物用GC-MS法定性、GC-NPD法定量检测,WinNorLin软件拟合房室模型并计算毒物代谢动力学参数。全程记录实验动物主要生命体征变化。结果氯胺酮和代谢物去甲氯胺酮在家兔体内的毒物代谢动力学过程均呈一级动力学特征,符合二室开放模型,氯胺酮毒物代谢动力学方程为ρt=121.760e-0.025t+0.980e-0.002t+4.579 e-0.021t,去甲氯胺酮毒物代谢动力学方程为ρt=640.919 e-0.03t+1.023 e-0.001t+9.784 e-0.031t。血液中氯胺酮质量浓度达峰时间为(40.950±12.098)min,血峰质量浓度为(9.015±1.344)μg/mL,消除半衰期为(430.370±28.436)min。给药后30～240 min内氯胺酮在血清和尿液中的质量浓度之间具有动态平衡的中度相关性。家兔给药后30min出现中毒症状,120min后渐恢复正常。结论建立的氯胺酮毒物代谢动力学方程和参数...     

液相色谱-串联质谱同时分析尿液中的大麻酚类和△^9-四氢大麻酸

目的建立LC/MS-MS同时检测尿液中Δ9-四氢大麻酚(THC)、大麻酚(CBN)、大麻二酚(CBD)和大麻主要代谢物Δ9-四氢大麻酸(THC-COOH)的方法.方法屎液样本经碱水解,加入氘代四氢大麻酸Δ9-d9-THC-COOH)内标,经V(正己烷)V(乙酸乙酯)=91提取,吹干,以100μL乙腈定容,利用LC/MS-MS方法进行分析.结果THC-COOH、CBN、THC和CBD的最低检测出质量浓度为0.2、0.4、1.0和2.0ng/mL;在阳性尿液中检出THC-COOH成分,质量浓度为335.9 ng/mL.结论所建立的方法简便快速、灵敏度高、专属性强,可满足检测尿液中THC、CBN、CBD以及大麻主要代谢物THC-COOH的要求.     

GC/MS、GC/NPD法检测血液中氯胺酮

目的利用GC/MS、GC/NPD与固相萃取（SPE）技术相结合,建立血液中氯胺酮的定性定量分析方法。方法选择4-苯基丁胺为内标,采用Bond-Elut Certify固相柱萃取、二氯甲烷：异丙醇：氨水（78∶20∶2,v/v/v）洗脱的固相萃取分离技术,比较不同pH体系、洗脱溶剂对回收率的影响,建立血液中氯胺酮的GC/MS、GC/NPD定性定量分析方法。结果以GC/NPD分析氯胺酮在6.0～5000ng/mL范围内线性关系良好,GC/MS-Scan定性检测限为20.0ng/mL。方法平均回收率达96.9%,标准偏差小于5%。结论此方法可满足氯胺酮毒品滥用者血液定性定量分析。     

家兔唾液与血液中氯胺酮浓度的相关性

目的研究家兔唾液中氯胺酮及代谢物去甲氯胺酮浓度与血药浓度的相关性。方法实验家兔分为氯胺酮灌胃组（6只）、静脉注射组（6只）和对照组（6只）,分别于染毒前、后不同时间点收集唾液和血液。采用气相色谱/质谱联用（GC/MS）全扫描定性、气相色谱（GC）定量分析样品中氯胺酮及去甲氯胺酮的浓度。采用双变量Pearson相关分析研究唾液中药物浓度和血药浓度的相关性。结果氯胺酮灌胃组和静脉注射组给药后各时间点氯胺酮及去甲氯胺酮在唾液和血液中的浓度相关系数（r）范围为0.80-0.95。结论氯胺酮及去甲氯胺酮在唾液和血液中的浓度均有良好的相关性,根据唾液药物浓度推断血药浓度可用于氯胺酮滥用的法医学鉴定。     

顶空固相微萃取气相色谱法快速检测尿液中氯胺酮

目的建立快速检测吸毒人员尿液中氯胺酮的顶空固相微萃取(HS/SPME)-GC/NPD的方法。方法样品瓶中加入尿液、氢氧化钠溶液、氯化钠,在95℃下加热搅拌,用聚二甲基硅氧烷SPME萃取头(100μm)顶空萃取20min,气相色谱氮磷检测器检测,外标法定量。结果尿液中氯胺酮浓度在0.2~20.0μg/ml范围内呈现线性关系(r2=0.9965),尿液添加1.0μg氯胺酮,平均回收率102.6%,RSD=3.39%(n=7),检测限1.13ng/ml(S/N=3)。结论建立的方法简单、快速、灵敏、准确,十分适合尿液等生物检材中氯胺酮的快速定性定量分析。     

SPE-HPLC/MS/MS法检测人唾液中地西泮及其代谢物

目的采用固相萃取-高效液相色谱-串联质谱法(SPE-HPLC/MS/MS)检测人唾液中地西泮及其代谢物。方法采用固相萃取法(SPE)处理唾液,HPLC/MS/MS法检测,MRM记录方式,保留时间和定性离子对定性,内标法和标准曲线法定量。结果地西泮及其代谢物去甲地西泮、去甲羟基西泮、去甲羟基地西泮葡萄糖醛酸苷(OG)、羟基地西泮葡萄糖醛酸苷(TG)的检测限在0.01ng/m L~0.5ng/m L之间,线性范围0.1ng/m L或0.5ng/m L~100ng/m L,回收率为84.9%~106%。口服5mg地西泮后15d内唾液中可检出地西泮及去甲西泮,但检出时间有个体差异,但去甲羟基西泮、TG和OG则不能检出。结论 SPE-HPLC/MS/MS检测法可应用于人唾液中地西泮及其代谢物的检测。人口服常量地西泮后唾液中可检出地西泮和去甲西泮,且检测窗口期较宽,但存在个体差异。     

Use of SPE and LC/TIS/MS/MS for rapid detection and quantitation of ketamine and its metabolite, norketamine, in urine

Ketamine (K) has become more and more popular for drug abuse in recent years. A lot of pre-treatment work such as extraction and derivatizing increase difficulties in the tests for ketamine in biological specimens. A rapid method to detect and quantitate ketamine and its metabolite norketamine in urine used deuterated dilution followed by solid phase extraction and liquid chromatography/TurboIonSpray/tandem mass spectrometry (LC/TIS/MS/MS) is described. Control recovery for both low and high concentrations can reach to 90%. Ten ketamine positive urines were examinated by this method. Concentrations ranged from 114 to 2925 ng/mL and from 453 to 9805 ng/mL for norketamine. The method was sensitive, specific, accurate and provided easy operation to detect and quantitate ketamine and its metabolites in urine.     

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。     

Assessment of age and sex by means of DXA bone densitometry: application in forensic anthropology

An effective way to reveal the history of drug abuse is to determine the parental drug and its metabolites in hair. Here, a quantitative HPLC-Chip-MS/MS method was developed for simultaneous measurement of ketamine and its metabolite norketamine in human hair. Ketamine and norketamine were extracted from hair by acid hydrolysis, and then enriched by organic solvent extraction. The chromatographic separation was achieved in 15 min, with the drug identification and quantification by a tandem mass spectrometer. The linear regression analysis was calibrated by deuterated internal standards with a R(2) of over 0.996. The limit of detection (LOD) and the limit of quantification (LOQ) for ketamine and norketamine were 0.5 and 1 pg/mg of hair, respectively. The standard curves were linear from the value of LOQ up to 100 pg/mg of hair. The validation parameters including selectivity, accuracy, precision, stability and matrix effect were also determined. In conclusion, this method was able to reveal the present of ketamine and norketamine with less hair from the drug abusers, and which had the sensitivity of ～1000-fold higher than the conventional method. In addition, the amount of ketamine and norketamine being detected in different hair segments would be useful in revealing the historical record of ketamine uptake in the drug abusers.     

尿中氯胺酮及其代谢物盘鉴和GC/MS/SIM测定

目的 研究尿中氯胺酮(KET)及其代谢物去甲基氯胺酮(NKET)的盘鉴(Disk SPE)。方法 用含有化学键合C18和强酸型强阳离子交换(SCX)基团的萃取柱SPEC.C18 AR/MP3萃取,加入萃取柱前的尿样用0.1mol/L磷酸盐缓冲溶液(pH 6)稀释,洗脱溶剂为含2％(v/v)氨水的乙酸乙酯;以2,4,6-三硝基甲苯(TNT)为色谱内标,GC/MS/SIM检测。结果 在加标量为0.5μg/mL、2μg/mL和6μg/mL的控制尿样中,KET和NKET的平均回收率分别为91.5％和79.9％,6次测定的RSD均为8.7％;线性范围0.02-8μg/mL,线性相关系数分别为0.9819和0.9964;检出限(S/N=3)分别为6ng/mL和4ng/mL;总离子色谱图背景低,杂质少。同一根萃取柱重复使用8次以上未见性能下降;嫌疑尿样中检出KET和/或NKET,和常规的液液萃取结果相符。结论 该方法适用于尿中KET和NKET的同时测定。     

Analysis of ketamine and norketamine in urine by automatic solid-phase extraction (SPE) and positive ion chemical ionization-gas chromatography-mass spectrometry (PCI-GC-MS)

Ketamine (KT) is widely abused for hallucination and also misused as a "date-rape" drug in recent years. An analytical method using positive ion chemical ionization-gas chromatography-mass spectrometry (PCI-GC-MS) with an automatic solid-phase extraction (SPE) apparatus was studied for the determination of KT and its major metabolite, norketamine (NK), in urine. Six ketamine suspected urine samples were provided by the police. For the research of KT metabolism, KT was administered to SD rats by i.p. at a single dose of 5, 10 and 20mg/kg, respectively, and urine samples were collected 24, 48 and 72 h after administration. For the detection of KT and NK, urine samples were extracted on an automatic SPE apparatus (RapidTrace, Zymark) with mixed mode type cartridge, Drug-Clean (200 mg, Alltech). The identification of KT and NK was by PCI-GC-MS. m/z238 (M+1), 220 for KT, m/z 224 (M+1), 207 for NK and m/z307 (M+1) for Cocaine-D(3) as internal standard were extracted from the full-scan mass spectrum and the underlined ions were used for quantitation. Extracted calibration curves were linear from 50 to 1000 ng/mL for KT and NK with correlation coefficients exceeding 0.99. The limit of detection (LOD) was 25 ng/mL for KT and NK. The limit of quantitation (LOQ) was 50 ng/mL for KT and NK. The recoveries of KT and NK at three different concentrations (86, 430 and 860 ng/mL) were 53.1 to 79.7% and 45.7 to 83.0%, respectively. The intra- and inter-day run precisions (CV) for KT and NK were less than 15.0%, and the accuracies (bias) for KT and NK were also less than 15% at the three different concentration levels (86, 430 and 860 ng/mL). The analytical method was also applied to real six KT suspected urine specimens and KT administered rat urines, and the concentrations of KT and NK were determined. Dehydronorketamine (DHNK) was also confirmed in these urine samples, however the concentration of DHNK was not calculated. SPE is simple, and needs less organic solvent than liquid-liquid extraction (LLE), and PCI-GC-MS can offer both qualitative and quantitative information for urinalysis of KT in forensic analysis.     

尸体脏器中安定、利眠宁及其代谢降解产物的毛细柱GC/MS鉴定

本文描述了尸体脏器中安定、利眠宁及其代谢降解产物二苯甲酮类、N-去甲安定、舒宁和脱氧利眠宁的毛细柱GC/MS鉴定方法,讨论了热解和代谢过程。     

尿中MDMA及其代谢物的GC和GC／MS分析

考察MDMA在人体内的代谢以及建立尿中MDMA和体内主要代谢物MDA的分析方法。尿样水解后经液-液提取处理,用GC／MS（EI、PCI）和GC／FID法分析。人摄入MDMA后尿中MDA和原体MDMA比约为0．10～0．14。GC／MS／SIM和GC／FID法的最低检出限为2ng／ml和50ng／ml,回收率大于85％,变异系数小于10％。该法简便快速、灵敏度高、结果可靠,可用于MDMA滥用者的尿样鉴定。MDA／MDMA浓度比可作为评判毒分结果的参考指标。