固相提取在毒物分析中的应用研究

应用GC、GC/MS研究了尿、肝、胃组织用液- 液提取法制样引入样品中的有机组分;考查了十几种树脂稳定性、吸脱性能.选用的GDX-403树脂作吸附剂具有稳定性好、吸附回收率高的特点,当苯巴比妥浓度为2.96mg/ml,吸附平衡回收率达98.8%.净化巴比妥类药物具有样品图谱清晰、无干扰、吸脱率再现性好的优点.     

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。     

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。     

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.     

体液中氟乙酰胺SPE-GC/MS检测

目的　利用GC/MS与固相萃取 (SPE)技术相结合 ,开发血和尿样中氟乙酰胺鼠药的GC/MS定量分析新方法 ,并用于实际案例检测。方法　选择乙酰胺为内标 ,通过比较不同固相柱的萃取效率和不同条件对回收率的影响 ,优化用于血和尿样中氟乙酰胺萃取的固相柱和提取条件 ,利用氟乙酰胺与乙酰胺质谱图的分子离子峰面积之比与氟乙酰胺浓度的定量关系 ,建立血和尿样中氟乙酰胺鼠药的GC/MS定量分析新方法。结果　用硅胶柱萃取 ,峰面积之比与氟乙酰胺浓度在 5 0～ 90 μg/ml范围呈线性关系 ,检测限为 1 0 μg/ml。血样中氟乙酰胺检测的平均回收率达 91 6% ,标准偏差小于 7 3 %。结论　此法对实际样品的测定证明可满足氟乙酰胺鼠药中毒的定性定量要求。     

吗啡及其代谢物在藏毒致死者体内分布初探

目的采用固相萃取、液相色谱一串联质谱（LC-MS／MS）检验方法,考察吗啡和葡萄糖醛酸吗啡（M3G）在一例体内藏毒致急性死亡者体内分布情况。方法提取死者心血、尿、胃内容物、肝、肾、脑等15种检材,经Waters HLB小柱固相萃取后,C18色谱柱分离,采用电喷雾电离（ESI）、多反应监测模式（MRM）检测目标化合物。结果所建方法在0．0l～101μg／mL浓度范围内线性关系良好,提取回收率大于75％。结果显示总吗啡含量（游离态＋结合态）在胃内容物中最高,其次是尿、‘肾,在心血、胃组织、肺和腺体中居中,脑组织和心脏含量最低。结论本例检验结果验证了胃内容物、尿液和肾脏等是该类中毒案件的理想检材,其分布规律也可作为体内毒品分析实验依据。     

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。     

固相吸附-GC／MS法检验鱼塘水中农药氰戊菊酯

目的建立鱼类中毒案件鱼塘水中微量农药的分析方法。方法取一定量的吸附剂GDX-403，加入到取出的一定量的鱼塘水中，经振荡吸附，分离出GDX-403，取有机溶剂解析、浓缩，进行GC／MS检验。结果回收率达90％，相对偏差59／5，1000mL水中农药氰戊菊酯检测限浓度在0．2ppb左右。结论该方法操作简便，准确、灵敏度高、可靠性好，可以进行实战推广。     

固相萃取技术分析生物检材中的鸦片成份

建立用国产GDX－403固相柱、GC／NPD毛细管气相色谱法，提取、净化生物检材中鸦片的方法。在1ml全血中添加1～4μg混合药物，回收率为79.2％～89.3％，RSD为2．61％～7．50％；在2g肝中添加2．5～10μg混合药物，回收率为72．9％～86．9％，RSD为2．27％～9．20％。该法回收率高，可用于检测人体内的微量鸦片成份。     

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.     

Detection of drugs using XAD-2 resin. I:Choice of resin, chromatographic conditions, and recovery studies

Amberlite XAD-2, a nonionic polystyrene divinylbenzene resin, was first used for the analysis of drugs in urine and a number of reports have described the development at optimal conditions for extraction, including type of resin columns, pH conditions, and eluting solvents. XAD-4 and XAD-7 resins were compared to the similarly structured XAD-2 resin and no significant advantage over the XAD-2 resin for drug screening was observed. A quantity of 5 to 6 g of resin was found to have sufficient capacity for the extraction of 200 ml of pentobarbital solution (1 mg/100ml). A column flow rate of approximately 15 ml/min (gravitational flow) was sufficient for analysis and slower rates were not more efficient. A mixture of ethyl acetate and 1,2-dichloroethane (3:2) was found to give best overall recovery (66 to 94%) of drugs, the resulting extracts being reasonably free of interfering substances. A pH value of 8.5 is recommended as optimum for comprehensive analysis of acidic and basic drugs. Recovery studies were conducted on spiked samples to determine drug losses occuring during various steps in the XAD-2 extraction procedure for four acidic (amobarbital, secobarbital, pentobarbital, and phenobarbital) and four basic (morphine, codeine, meperidine, and methadone) drugs. A relatively small amount (0 to 5%) of the drugs was not adsorbed by the resin and amounts varying from 6 to 40% failed to be desorbed by the eluting solvent. Additional losses occurred during the removal and analysis of TLC spots. Recovery of drugs from aqueous solutions analyzed with the XAD-2 resin were compared to recoveries reported in the literature with other XAD-2 resin methods for the extraction of drugs from urine. Recovery of phenobarbital, morphine, and codeine improved by 4 to 23% while recoveries of amobarbital, pentobarbital, secobarbital, methadone, and meperidine were 4 to 28% less efficient when compared to literature data.     

柱切换HPLC法分析尿中吗啡和O^6—单乙酰吗啡的研究

目的建立用柱切换HPLC技术分析尿中吗啡和06-单乙酰吗啡的方法.方法尿样用硼砂缓冲液(pH9.2)稀释后进入预处理柱(YWG-C18,33mm×5.0mm,10μm),用H2O洗去杂质,再用CH3OHH2O(6040)将被分析组分洗脱进入分析柱(Lichrospher(R)100CN,125mm×4.0mm,5μm),分析流动相为CH3OH磷酸盐缓冲液(pH6.86)=2278.紫外检测器波长为286nm.结果尿中吗啡和06--单乙酰吗啡的线性范围分别为50～1 600n/ml和100～1 600n/ml.吗啡和O6--单乙酰吗啡的精密度均小于4%.吗啡和O6-单乙酰吗啡的检测限均为40n/ml.结论用CSHPLC测定尿中吗啡和O6-单乙酰吗啡,方法准确、灵敏、快速、简便.     

SPME结合衍生化检测尿液中吗啡成份

目的　建立尿液中吗啡检验的方法。方法　利用固相微萃取头富集尿液中吗啡 ,经醋酸酐衍生后进行GC/MS分析。结果　利用此方法可检测到吗啡含量为 5ug/ml的尿样。结论　该方法可用于吸毒人员尿液中吗啡的快速检验。     

固相萃取／LC-MS／MS测定尿液中吗啡类药物

目的 建立尿液中吗啡类药物的固相萃取／LC—MS／MS方法。方法采用OASIS MCX3cc（60mg）固相萃取柱进行提取,应用LC—MS／MS方法进行检测,运用保留时间和MRM方式对尿液中吗啡类药物及其代谢物进行定性定量分析。结果磷酸盐缓冲液pH4．0时,海洛因、6-MAM、可待因、吗啡、M3G的固相萃取回收率分别达64．33％-70．21％,96．95％～117．57％,83．60％～123．63％,68．82％～91．03％,94．64％～107．33％;最低检测限（LOD）分别为5、10、5、5、2pg,线性范围0．005～10μg／mL;相关系数分别为0．9998、0．9958、0．9992、0．9994、0．9997。结论本文所建方法,适用于尿液中吗啡类药物的分析。     

生物试样中巴比妥类药物固相提取柱上衍生化GC/MS分析

建立生物试样中常见巴比妥类药物的固相提取和柱上衍生化GC／MS分析法。将预制的血或肝分别在pH6．0和pH2.2的条件下过预活化的GDX－403吸附小柱，再用缓冲浪和蒸馏水各4ml顺序洗柱。最后用4ml丙酮／氯仿（1：1）溶剂洗脱样品，离心弃除水相，80℃挥至近干，用50μl乙醇定溶、取净化的样品2～4μl挥至近干，加20μl0．2molTMAH衍生化试剂，直接进样0.5μl，柱上衍生化GC／MS（GC）分析。在试验条件下，当血和肝分别添加2．0μg和5．0μg混合药物，回收率≥80％，相对标准差（RSD）优于±10％，检测限优于5ng（信／噪比≥2）。该法能有效地排除类脂物和组胺的干扰，可用于治疗量级药物分析和婴幼儿中毒案检验。     

尿中氯喹定性定量分析方法的研究

建立了人尿中氯喹的定性定量分析方法,2ml尿样用2ml×2环己烷：乙酸乙酯（8：2）提取净化后,60℃水浴室气吹干,残留物定容溶解后,气相色谱分析,氯喹的保留时间为9．44min。方法最低检测限为200ng／ml,回收率为87．0％,RSD＝7．9％（n＝5）,在0～50μg／ml浓度范围内,有良好的线性关系：A＝1778．9＋13686C,r＝0．999。方法同时可用于血中氯喹的分析。附一例应用报告,测得尿中氯喹的含量为0．745mg／ml,血中氯喹的含量为3．68μg／ml。尿液中同时检出氯喹的N－去单已基代谢物。定性结果经质谱法验证。     

静松灵的检验

本实验建立了用固相萃取(SPE)、薄层色谱和气—质联用法(GC-MS)分析血、尿、肝中静松灵的方法,在1g尿、血、肝中添加0.01mg药物、回收率分别为96.1%、90%、67%.该方法简单快速准确,经动物实验及案件鉴定验证,效果较好.     

GC/MS法测定血液中的米氮平

目的采用液液提取-GC/MS法检测血液中米氮平。方法用0.1mol/L NaOH溶液调节血液pH值为8～9,以二氯甲烷∶环己烷(4∶6)混合溶剂作为提取溶剂,液液提取法提取血液中米氮平,外标法气相色谱/质谱联用仪定性定量分析。在2mL血中分别添加标准米氮平1、0.5、0.1μg,进行回收率测定。结果血液中添加米氮平的平均回收率为91.5%,方法的工作曲线为Y=1E+06X-16206,相关系数r=0.999 7,线性范围0.5～10μg/mL,最小检出限0.1μg/mL(S/N=46)。结论本文方法操作简便,适合于血液中米氮平的定性、定量分析,可在实际捡案中选择使用。     

Validation of a high performance liquid chromatographic method for alpha amanitin determination in urine

The objective of the present study was to develop and validate a liquid chromatographic method with electrochemical detection to measure alpha amanitin concentrations in urine after sample pretreatment with double mechanism (reversed phase/cation exchange) solid-phase extraction cartridges. The urine samples (10 ml) were purified and concentrated to 1 ml with elimination of matrix contaminants. The extracts were then separated by isocratic reversed-phase chromatography using a C18 column (4.6 mm×25 cm) with a mobile phase composed of 0.005 M phosphate buffer (pH 7.2) and acetonitrile (90:10). Coulometric detection was performed by applying an oxidation potential of +500 mV to a porous graphite electrode in a low-volume analytical cell. The limit of quantitation was 10 ng/ml with a signal-to-noise ratio=25. The linearity studied on spiked urine was satisfactory (r=0.9966) from 10 ng/ml to 200 ng/ml. The average extraction recovery of alpha amanitin was 78%, determined using spiked urine samples ranging from 10–300 ng/ml. The intra-assay precision was checked at 10, 50 and 100 ng/ml levels (n=10) in spiked urine samples, with resulting coefficients of variation of 3.6%, 2% and 1.5%, respectively.     

SPE-LC-MS/MS法测定尿液与血液中海洛因3种代谢物

目的采用SPE-LC-MS/MS方法,同时检测尿液与血液中海洛因主要代谢物3-β-D-葡萄糖醛酸吗啡(M3G)、吗啡和O6-单乙酰吗啡(O6)。方法采用BAKERBONDTMspe Octadecyl(C18)进行提取,应用LC-MS/MS方法检测并通过MRM及内标法进行量化。结果尿液中M3G、吗啡、O6-单乙酰吗啡的最低检测限(LOD)分别为1.24pg、6.71pg、0.47pg;回收率依次为82.25±12.25%、93.75±13.25%、88.70±11.90%。血液中M3G、吗啡、O6-单乙酰吗啡的最低检测限分别为1.50pg、8.21pg、0.52pg。回收率依次为89.85±21.15%、73.70±17.90%、90.10±3.90%。结论本文所建方法同时适用于尿液与血液中海洛因主要代谢物M3G、吗啡、O6-单乙酰吗啡的提取、净化、分析。