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。     

液相色谱-质谱联用法测定人全血中灭多威

目的建立一种简便的测定人全血中灭多威的液相色谱-质谱联用法。方法样品处理采用液-液乙酸乙酯萃取方法。色谱柱为Zorbax SB-C18(2.1mm×50mm,5μm),流动相为乙腈-0.1%甲酸,梯度洗脱,流速为0.5mL/min,柱温40℃。采用ESI离子源,MRM离子方式监测。结果灭多威在0.05~2.0μg/mL浓度范围内线性良好(r0.995)。灭多威的方法回收率均在90%~108%的范围内,日内、日间RSD均小于15%。结论本方法可简单、高效地检测全血中灭多威浓度。     

超声辅助固相萃取-HPLC法测定不同地区缴获鸦片样品中生物碱

目的本文建立了超声辅助固相萃取-HPLC法定量测定缴获鸦片样品中吗啡、可待因、蒂巴因、罂粟碱、那可汀5种生物碱并同时检出6种未知化合物的方法。采用主成分分析法区分不同地区缴获的鸦片样品。方法采用超声辅助固相萃取法提取生物碱,XDB-C18（5μm×4.6mm×250mm）色谱柱,流动相为10mM的1-庚烷磺酸钠（pH 3.2）和乙腈,梯度洗脱,紫外检测器检测。结果该方法的定量限为0.58~2.78 mg/kg。添加水平在0.2～1.5mg/mL范围内,平均加标回收率为81.0%～99.2%,相对标准偏差为1.6%～5.0%。结论本方法重现性好、定量准确,满足定量检测鸦片中生物碱含量并对不同产地鸦片进行分类的需要。     

血液和尿液中多种头孢菌素的HPLC分析

目的 建立检测血液和尿液中头孢唑啉、头孢拉定、头孢哌酮和头孢噻肟的高效液相色谱法,并考察方法的适用性.方法 以乙酰苯胺为内标,血液和尿液用乙腈直接沉淀蛋白,采用Agilent Zorbax SB-Aq(250min×4.6mm,5 μm)色谱柱,流动相选用水(含0.12%三乙胺和0.12%乙酸)和乙腈的混合溶剂梯度洗脱,流速1 mL/min,检测波长254nm.结果 4种头孢菌素分离完全,工作曲线相关性均良好(r>0.9990),检出限为0.01μg/mL,回收率均不低于81.2%.结论 所建立的方法对血液和尿液中以上4种头孢菌素的检测快速、简便、准确,适用于实际检案中头孢菌素类药物的分析.     

在线固相萃取-超高效液相色谱-串联质谱法同时测定血液样品中12种卡西酮类毒品

目的 建立一种同时测定血液样品中12种卡西酮类毒品的在线固相萃取-超高效液相色谱-串联质谱法。方法 血液样品用乙腈沉淀蛋白，经离心、稀释、过滤后上样，采用PLRP-S在线固相萃取柱（2.1mm×12.5mm,15～20μm）富集纯化，Poroshell 120 EC-C18色谱柱（3.0mm×150mm,2.7μm）进行分离，在线固相萃取柱以乙腈-5%（体积分数）甲醇作为流动相进行流速1.0 mL/min的梯度洗脱，色谱柱以5 mmol/L乙酸铵缓冲液[含0.1%（体积分数）甲酸]-乙腈作为流动相进行流速0.4m L/min的梯度洗脱。离子源为电喷雾离子源，采用多反应监测模式进行测定。结果 12种卡西酮类毒品线性关系良好，相关系数均大于0.998，方法检出限为0.1～0.5ng/mL，定量限为0.3～1.5ng/mL。12种卡西酮类毒品在3个不同质量浓度条件下的回收率为70.9%～108%，日内精密度和日间精密度分别为1.5%～8.9%、5.1%～44.5%(n=6)。结论 该方法操作简单方便、样品需求量少、灵敏度高、检出限低，可用于血液样品中卡西酮类毒品的测定。     

兔尿中敌鼠及其代谢物的HPLC-DAD分析

目的 尿中敌鼠及其代谢物的HPLC分析方法研究。方法 用CN柱和SAX柱的固相萃取(SPE)技术分离提取兔尿中的敌鼠及其代谢物,用香豆素作内标,用HPLC-DAD方法进行fenxi。色谱柱:Hypersil BDSC_(18)(150×4.6mm):保护柱:Phenomenex C_(18) (ODS,4×3.0mm,Octadecyl;流动相A:0.5％离子对A水溶液,B:0.5%离子对A甲醇液,梯度程序洗脱;DAD检测波长为311nm。结果 在中毒的兔尿中检出11种敌鼠代谢物。结论 此方法简单、准确、快速。     

固相萃取-气相色谱法检测全血中佐匹克隆

目的采用固相萃取-气相色谱法检测全血中佐匹克隆。方法采用Oasis HLB固相萃取柱对样品进行前处理,去离子水、0.5%氨水-甲醇/水((V/V 40∶60)溶液先后淋洗,二氯甲烷/异丙醇(V/V 75∶25)洗脱后进行GC/NPD检测。结果血液中佐匹克隆在50～5 000ng/mL范围内线性良好(R2=0.998 8),平均萃取回收率为96.9%,检出限为30ng/mL,日内RSD为2.1%～5.7%,日间RSD为3.3%～6.2%,结论固相萃取-气相色谱检测法灵敏度高,重现性好,可在血液中佐匹克隆的检测中选用。     

高效液相色谱法测定卡西酮

目的建立卡西酮的高效液相色谱检测方法。方法采用UPLC-DAD分析方法。分析柱:Agilent ZorbaxSB-Phenyl柱(250mm×4.6mm,5μm),流动相为三氟乙酸(pH 3.5)∶乙腈为85∶15,流速0.2mL/min,检测波长254nm。结果卡西酮在0.5～1 000μg/mL浓度范围内线性关系良好R2=0.999 4,日内与日间保留时间和峰面积的标准偏差(RSD)均<1.06%,检出限为0.068μg/mL,平均回收率95.9%。结论本方法峰形好,分离度好,线性范围良好,回收率高,适用于刑事案件中卡西酮的定性定量分析。     

UPLC-MS/MS法同时检测全血中16种除草剂

目的建立全血中16种除草剂的超高效液相色谱-串联质谱(UPLC-MS/MS)同时检测方法,为除草剂中毒案事件及其他刑事案件血液中该16种除草剂的检验鉴定提供依据。方法取200μL的血液,加入800μL乙腈-水(体积比80/20),进行蛋白沉淀后,采用Acquity BEH C18(2.1mm×100mm,1.7μm)色谱柱,以水(5mmol/L的甲酸铵,0.1%的甲酸)-乙腈为流动相进行梯度洗脱,采用电喷雾离子源(ESI)、多反应监测(MRM)正离子模式对16种化合物进行检测。结果在1~200ng/mL范围内线性关系良好,R2均大于0.996;基质效应(ME%)为85.2%~104.4%,相对标准偏差(RSD%)为0.72%~4.84%;仪器检出限(IDLs)为0.2~2 ng/mL(信噪比S/N≥3),方法检出限(MDLs)为0.5~3ng/mL(信噪比S/N≥3),最低定量限(LOQs)为1~7ng/mL(信噪比S/N≥10)。结论本实验建立的全血中16种除草剂同时检验方法,前处理简便快捷、回收率高、精密度好、方法检出限低,可作为该16种除草剂中(投)毒案件的检验方法。     

LC-MS／MS测定尿液中可卡因及其代谢物苯甲酰爱康宁

目的建立尿液中可卡因(cocaine,COC)及其代谢物苯甲酰爱康宁(benzoylecgonine,BZE)的液相色谱-串联质谱分析方法。方法尿液经固相萃取后,用AllurePFP丙基柱分离,以V(甲醇):V(20mmol/L乙酸胺和0.1%甲酸的缓冲溶液)=80∶20为流动相,采用二级质谱多反应监测模式检测COC和BZE。按10mg/kg的剂量对豚鼠腹腔注射可卡因,给药后收集7d尿液。结果尿液中COC和BZE在2.0～100ng/mL质量浓度范围内线性关系良好(r=0.9995),最低检测限(LOD)为0.5ng/mL;回收率大于90%;日内和日间精密度均小于6%;豚鼠尿液中主要检测目标物是BZE,且BZE检测时限也较COC长。结论所建方法灵敏度高,选择性好,适用于尿液中可卡因和苯甲酰爱康宁的检测。     

HPLC simultaneous determination of glycerol and mannitol in human tissues for forensic analysis

A method for simultaneous determination of glycerol and mannitol in various human tissues was devised and for this we used high-performance liquid chromatography (HPLC). Specimens were homogenized in a mixture of chloroform and methanol, phosphate buffer (pH 7.0) and pentaerythritol (IS) solution. After centrifugation, an aliquot of the aqueous layer was evaporated to dryness and derivatized with p-nitrobenzoyl chloride at 50 degrees C for 1h, then applied to HPLC with analytical conditions of: column, CAPCELL PAK C18 MG (250 mm x 3.0 mm i.d., 5 microm, Shiseido Co. Ltd., Tokyo, Japan); column temperature, 1-2 degrees C; mobile phase, 75% acetonitrile-distilled water containing 0.05% trifluoroacetic acid, 0.05% heptafluoro-n-butyric acid and 0.1% triethylamine; flow rate, 0.5 ml/min; wavelength, 260 nm. Calibration curves for both substances were linear in concentration ranges from 1 to 500 microg/0.1g and correlation coefficients exceeded 0.99. The relative standard deviation (R.S.D.) of the method was evaluated at concentrations of 10 and 100 microg/0.1g, and ranged from 0.84 to 10.6%. Using this method, we determined the regional distribution levels of glycerol and mannitol in various tissues from an autopsied brain dead man.     

固相萃取毛细管区带电泳法测定全血中甲基苯丙胺

目的建立全血中甲基苯丙胺的固相萃取毛细管区带电泳法检验方法。方法血液采用OasisHLB固相萃取小柱直接萃取,BeckmanP/ACEMDQ毛细管电泳仪区带法分析。结果回归方程y=0.0083x-0.0164,线性范围5～75μg/mL(r=0.994),日内精密度RSD=2.22%,日间精密度RSD=5.31%,盐酸甲基苯丙胺质量浓度为25.0μg/mL的全血相对回收率(91.63±2.5)%。电泳分离良好,空白无干扰。结论本方法操作简便,适用于全血中甲基苯丙胺的检验。     

生物检材中乌头碱的LC-MS/MS快速分析

目的应用高效液相色谱-质谱法对生物检材中乌头生物碱等有毒成分进行快速分析。方法取全血样品经乙腈-甲醇(5:1 v/v)提取,使用Agilent Zorbax SB C18(2.1 mm×50 mm,1.8μm)色谱柱,以0.1%甲酸溶液-乙腈(60:40 v/v)为流动相等度洗脱。在多反应监测模式下测定全血样品中乌头生物碱等有毒成分。结果乌头碱、次乌头碱和中乌头碱的保留时间为0.73 min、0.77 min和0.63 min;用于定量分析的离子对分别为m/z 646.4→586.4(乌头碱)、616.1→556.5(次乌头碱)和632.4→572.1(中乌头碱)。乌头碱在0.1~250 ng/m L内线性关系良好,相关系数(r)≥0.9987,最低检出限0.1ng/m L,精密度考查其变异系数(CV)5.42%(n=6),血液中乌头碱提取回收率不小于90%。结论本文建立的高效液相色谱-质谱法快速、简便、灵敏,适用于天然药毒物检验。     

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.     

全血中苯骈二氮杂类药物的胶束电动色谱法分析

目的建立用固相萃取胶束电动毛细管色谱法测定人体全血中苯骈二氮杂艹卓类药物的方法。方法全血以Oasis小柱提取,以克仑特罗为内标,采用未涂层毛细管(75μmID×50.2cm,有效分离长度为40cm),缓冲液为30mmol/LSDS→15mmol/L硼砂→15mmol/L磷酸盐(pH8.2)→18%甲醇。进样条件:压力进样0.5psi×10s,分离电压为25kV,柱温25℃,检测波长为230nm。结果本法分离效率高,9种苯骈二氮杂艹卓类药物的最低检测浓度为5～50ng/ml;血药浓度的线性范围为0.02～1.6μg/ml,日内、日间精密度<12%。结论本法简便、高效、可靠。     

Forensic and chemical determination of methane in cadaveric samples

Conditions were elaborated for the determination of methane, through gas chromatography, in blood and the lung by using a 2% methanol solution as an internal standard and with the below gas-chromatography column being applied: dimensions--200 x 0.3 cm, 15% di-2-ethylhexyl sebacate on Dinochrome II (0.16 x 0.21 mm) at 110 0 degree C. The detector is of the flame-ionization type. The normal content of methane was determined for blood and for the lung--0.122 +/- 0.0074 microgram/ml and 0.20 +/- 0.04 microgram/g, respectively. Biological samples from cadavers of other persons who died due to trauma were suggested for use as controls. The method was used to examine expertise objects, blood and the lung from the cadavers of peoples who died in fire.     

Screening and determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry

Benzodiazepines are one of the most widely prescribed drugs for the treatment of a wide spectrum of clinical disorders. They are used as anticonvulsants, anxiolytics, hypnotics or muscle relaxants with different duration of action. In this paper, a simple and sensitive method for the determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry (GC/MS) is described. The drugs spiked in whole blood were extracted with an Oasis HLB solid-phase extraction cartridge (Waters), which contains a copolymer designed to have a hydrophilic-lipophilic balance. GC/MS analysis was performed using a Shimadzu QP-5000 equipped with a BPX5 capillary column (15 mx0.32 mm I.D., film thickness 0.25 microm, SGE). Nineteen benzodiazepines and two thienodiazepines were well separated from each other on their SIM chromatograms and also on the TIC with the exception of oxazolam to cloxazolam separation. The blank extract from whole blood gave no peaks that interfered with all benzodiazepines and thienodiazepines on the chromatogram. The calibration curves for selected benzodiazepines with fludiazepam as an internal standard showed excellent linearity over the concentration range 5-500 ng/ml blood with a correlation coefficients of >0.995. The detection limits ranged from 0.2 to 20 ng/ml blood. The method is simple and sensitive for the determination of benzodiazepines in whole blood and seems to be useful in the practice of forensic science.     

血中毒鼠强的固相萃取和GC-NPD法测定

目的 研究人全血中毒鼠强的固相革取(SPE)。方法 用Bond Elute C18固相萃取柱萃取,GC-NPD检测,以甲基对硫磷为色谱内标(CS)。结果 全血中加标0.5μg/ml,毒鼠强回收率为92.5％,变异系数2.3％(CV,n=4)。在0.5～10μg/ml的浓度范围内,线性相关系数为0.9994。检出限(S/N=3)和定量限(S/N=10)分别为6ng/ml和20ng/ml。同一根萃取柱连续使用6次未见性能明显下降(CV=3.6%)。结论 本文方法适用于毒鼠强中毒的全血测定。