分子印迹固相萃取法在血中苯丙胺类毒品分析中的应用

目的建立分子印迹固相萃取（MISPE）、GC/MS分析方法,用于血液中苯丙胺类毒品检测。方法 10mmol/L醋酸铵缓冲液（pH8.0）4倍稀释空白添加血液,1mL甲醇,1mL10mmol/L醋酸铵缓冲液（pH8.0）活化苯丙胺类分子印迹固相萃取柱;2×1mL去离子水、1mL60%的乙腈去离子水、1mL1%醋酸乙腈洗涤杂质;2×1mL1%甲酸/甲醇洗脱,洗脱液挥干定容,经GC/NPD、GC/MS分析检测。结果各种苯丙胺类毒品回收率均在90%以上,在20～5 000ng/mL浓度范围内线性关系良好,r2为0.995 7～0.998 9,LOQ在16～30ng/mL之间,LOD在8～15ng/mL之间。结论本方法回收率高,净化效果显著,稳定性好,杂质干扰少,可用于血液中低浓度苯丙胺类毒品的分析检测。     

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。     

固相萃取/气相色谱法检测全血中的扎来普隆

目的建立全血中扎来普隆的固相萃取/气相色谱检测方法。方法采用Oasis HLB固相萃取柱,5%甲醇水淋洗,二氯甲烷洗脱,GC/NPD定量检验,GC/MS定性检验,并对检测条件进行考察。结果扎来普隆与血中杂质分离良好,平均萃取回收率达89%以上,最低检出限20ng/mL,50～2000ng/mL内线性良好,相关系数R2=0.9945,日内RSD为3.1%～4.2%,日间RSD 5.8%～6.7%。结论该方法操作简便快速,提取回收率高,重现性好,提取物杂质少,可用于实际案件的检验。     

UPLC-MS/MS法检测血液中的磺胺嘧啶

目的建立一种采用UPLC-MS/MS检测全血中磺胺嘧啶的方法。方法以HLB固相萃取柱提取血样,应用UPLC色谱法分离,MS/MS检测分析。结果该方法具有浓缩富集的效果,使血液中磺胺嘧啶的回收率高于102%,线性范围为10～1 000ng/mL,检测限0.21ng/mL(S/N≥3)。结论该方法灵敏度高,分析速度快,操作简便,可用于全血中磺胺嘧啶的定性、定量分析检验。     

固相萃取-液相色谱-质谱法检验人全血中的地塞米松

目的采用固相萃取结合液相色谱-质谱法检验人全血中地塞米松。方法采用SPE提取全血,用UPLC-MS/MS方法测定,采用ESI离子源,MRM方式监测。结果地塞米松的检出限为0.05ng/mL,线性范围1ng/mL~100ng/mL,方法回收率大于78.1%,日内、日间精密度均小于15%。结论 SPE-UPLC/MS/MS检验人全血中地塞米松的方法简单、高效,可应用于人全血中地塞米松的检验。     

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。     

离子交换柱法萃取血样中4种巴比妥类药物

目的建立血样中4种常见巴比妥类药物的固相萃取方法。方法采用Oasis MAX固相萃取柱对样品进行预处理,以2%氨水淋洗2,%甲酸-甲醇洗脱,GC/NPD定量检验,GC/MS定性检验。结果血样中4种常见巴比妥类药物平均萃取回收率达80%以上,以信噪比10/1计,最小定量限为1～2μg/mL,以信噪比3/1计,检出限为0.5～1μg/mL,在1～50μg/mL范围内,线性相关系数均在0.99以上。结论本文固相萃取方法检出限低,回收率高,选择性强,可应用于血样中4种常见巴比妥类药物的检测。     

SPE—GC／MS、GC／NPD法检测血液中苯丙胺类毒品

目的利用GC／MS、GC／NPD与固相萃取（SPE）技术相结合,建立血液中苯丙胺类毒品的定性定量分析方法。方法采用Bond—ElutCerti{y固相柱、甲醇淋洗、二氯甲烷／异丙醇／氨水（78／20／2）洗脱固相萃取分离提取,比较了不同PH体系、稀释状态、洗脱溶剂对提取回收率的影响,建立血液中苯丙胺类毒品的GC／MS、GC／NPD定性定量分析方法。结果以GC／NPD分析AM、MA、MDA和MDMA浓度在15ng／mL-2000ng／mL、10ng／mL～1600ng／mL、20ng／mL-3000ng／ml、20ng／mL-3000ng／mL范围内线性关系良好,AM、MA、MDA和MDMA的检测限分别为10ng／mL、8ng／mL、15ng／mL、15ng／mL,方法平均回收率大于85％,标准偏差小于5％,GC／MS-Scan检测限分别为40ng／mL、32．0ng／mL、60．0ng／mL、60．0ng／mL。结论此方法可满足苯丙胺类毒品滥用者的血液定性定量分析。     

离子液体分散液相微萃取-HPLC检测血浆中溴敌隆

目的建立离子液体分散液相微萃取-HPLC检测血浆中溴敌隆的方法。方法以1-己基-3-甲基咪唑六氟磷酸盐([HMIM]PF6)为萃取剂,甲醇为分散剂,提取血浆中的溴敌隆,用HPLC分析其含量;考察了萃取剂和分散剂体积、样品pH值、NaCl浓度、萃取时间和离心时间等因素对萃取效率的影响。结果优化的溴敌隆分散液相微萃取操作条件为:萃取剂[HMIM]PF650μL,分散剂甲醇100μL,样品pH值5.0,萃取时间5min,离心时间8 min。方法的线性范围:0.01～5.0μg/mL,检测限:1.1 ng/mL(S/N3)。该法低、中、高浓度的平均加标回收率分别为76.4%,82.6%,92.1%,RSD分别为4.17%,2.99%,1.67%(n=6)。结论本方法检测血浆中的溴敌隆,简便快速、准确实用,满足中毒检测及临床诊断治疗的需要。     

使用LC-MS/MS法检验人体毛发中的西地那非成分

目的建立人体毛发中西地那非成分的LC-MS/MS检验方法,并应用于实际案例。方法色谱柱为Thermo Hyperil Gold(50mm×2.1mm, 1.9μm),流动相为甲醇-5mM醋酸铵-0.1%甲酸,采用ESI源正离子多反应检测(MRM)扫描分析。结果西地那非的检出限为0.1ng/mL,定量限为0.2ng/mL,在0.2ng/mL～100ng/mL范围内线性良好,线性系数0.9999,回收率为86.7%～95.5%。结论该方法快速、灵敏、高效,可适用于实际案件人体毛发中西地那非成分的检验。     

Determination of bromadiolone and brodifacoum in human blood using LC-ESI/MS/MS and its application in four superwarfarin poisoning cases

Superwarfarin poisoning is a growing health problem. A sensitive and reproducible LC-ESI/MS/MS (liquid chromatography electrospray ionization tandem mass spectrometry) method was developed and validated for the determination of bromadiolone and brodifacoum, the most commonly used superwarfarins, in human blood using warfarin-D5 as an internal standard. Bromadiolone and brodifacoum were extracted from whole blood samples by liquid-liquid extraction with ethyl acetate. Multiple-reaction monitoring (MRM) was used to detect bromadiolone and brodifacoum using precursor→product ion combinations of m/z 525→250 and 521→135, respectively. The calibration curves were linear (r(2)=0.9999) in the concentration range of 0.5-100.0ng/mL for bromadiolone and brodifacoum, with a lower limit of detection of 0.1 and 0.2ng/mL, respectively, in whole blood. This method detected trace levels of bromadiolone and brodifacoum in whole blood samples and can be used in the diagnosis of poisoned human beings.     

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。     

全血中氯霉素的固相萃取方法

目的建立全血中氯霉素的固相萃取方法。方法在空白血中添加标准氯毒素,采用HLB、MCX固相柱萃取,HPLC法测定了线性范围、精密度、回收率。结果回收率分别为：HLB为69.1%,RSD为6.21%,MCX为70.1%,RSD为4.34%。标准工作曲线Y=18.1094X-0.4822,相关系数r=0.9999,线性范围1.0-20.0ug/L,最小检出量3ng。标准添加工作曲线Y=-2.1165X＋14.0459,相关系数r=0.9996,最小检出限0.5μg/mL（S/N=10∶1）。结论此二种萃取柱均可用于氯霉素的固相萃取。     

Simultaneous Determination of 13 Anticoagulant Rodenticidesin Human Blood by Liquid Chromatography–Tandem Mass Spectrometry and its Application in Three Poisoning Cases

Anticoagulant rodenticides are widely used for rodent control around the world. A rapid and sensitive method was developed and validated for the simultaneous determination of 13 anticoagulant rodenticides (coumafuryl, pindone, valone, warfarin, coumatetralyl, coumachlor, diphacinone, dicumarol, chlorophacinone, bromadiolone, difenacoum, flocoumafen, and brodifacoum) in human blood by liquid chromatography–tandem mass spectrometry. After liquid–liquid extraction, the anticoagulant rodenticides were separated on an Eclipse Plus C18 column. Linearities were observed for each analyte in blood ranging from 0.5 to 50 ng/mL, with correlation coefficients over 0.99. The limits of detection ranged from 0.01 to 0.2 ng/mL, and the limits of quantification were 0.5 ng/mL for all analytes. The intraday and interday precisions were <15%, and accuracies ranged from 80.3% to 111.0%. This validated method with high sensitivity has been applied in three anticoagulant rodenticide poisoning cases and has been used successfully in monitoring blood concentrations for months.     

GC-MS/MS测定血液中巴比妥类安眠药物

建立GC-MS/MS测定血液中巴比妥类安眠药物的分析方法。方法通过固相萃取提取并富集血液样品中常见巴比妥类安眠药物,采用离子阱二级质谱定性并定量检测其含量,并优化萃取溶液pH值与气相色谱/二级质谱联用分析条件,对巴比妥类安眠药物进行定量分析。结果巴比妥类安眠药物检出限为0.04μg/mL～0.10μg/mL,回收率为80.3%～92.6%。结论该方法高效、简单,灵敏度高,可用于血液中巴比妥类安眠药物同时定性定量检测。     

Fatal brodifacoum rodenticide poisoning: autopsy and toxicologic findings.

This report details the pathologic and toxicologic findings in the case of a 15-year-old girl who deliberately and fatally ingested brodifacoum, a commonly used rodenticide. The mechanism of death, massive pulmonary hemorrhage, has not been previously reported. Brodifacoum was quantitated in liver, spleen, lung, brain, bile, vitreous humor, heart blood, and femoral blood using HPLC with fluorescence detection. The highest brodifacoum concentrations were detected in bile (4276 ng/mL) and femoral blood (3919 ng/mL). No brodifacoum was detected in brain or vitreous humor. A brodifacoum concentration of 50 ng/g was observed in frozen liver while formalin fixed liver exhibited a concentration of 820 ng/g. A very high blood:liver brodifacoum concentration ratio suggested acute poisoning but the historical and pathologic findings suggested a longer period of anticoagulation. Though most cases of brodifacoum poisoning in humans are non-fatal, this compound can be deadly because of its very long half-life. Forensic pathologists and toxicologists should suspect superwarfarin rodenticides when confronted with cases of unexplained bleeding. Anticoagulant poisoning can mimic fatal leukemia or infectious diseases such as bacterial sepsis, rickettsioses, plague, and leptospirosis. A thorough death scene investigation may provide clues that a person has ingested these substances.     

血清中盐酸曲马多的膜式固相萃取及GC／MS／SIM测定

目的建立血清中盐酸曲马多的膜式固相萃取(SPE)气-质联用分析方法。方法1mL血清用2mL0.1mol/L磷酸盐缓冲液(pH6)稀释后,用含有C18和强酸型强阳离子交换基团的SPECC18AR/MP3固相萃取柱萃取,洗脱液为含2%氨水的乙酸乙酯;选择SKF525作为内标物,用GC/MS/SIM定量测定。结果在加标量为0.1、0.2和0.5μg/mL的血清样品中,盐酸曲马多的回收率分别为98.9%、92.5%和84.8%,5次测定的RSD分别为3.2%、8.7%和10.9%;线性范围为0.1～4μg/mL,多项式回归相关系数r2=0.9939;检出限为21ng/mL;同一根萃取柱,连续使用5次,没有出现堵塞和污染,回收率及RSD未见下降。结论本方法适合于法医毒物分析。     

HPLC-ESI-MS/MS法分析肝组织中4种杀鼠剂

目的建立测定杀鼠迷、杀鼠灵、溴敌隆和大隆4种杀鼠剂的高效液相色谱电喷雾质谱联用检测方法。方法采用ZORBAX sb-aq,150mm×2.1mm×3.5μm(带预柱);柱温:30℃;流动相:甲酸(0.1%)甲醇和甲酸(0.1%)水溶液梯度条件;流速:0.4 mL/min;在负离子模式下,通过电喷雾电离(ESI),多反应离子监测(MRM)测定,外标法定量分析4种杀鼠剂,并测试方法的基质抑制作用。结果4种鼠药在MRM筛选条件下,最小检出均在40pg以下;单个鼠药线性相关系数均在0.999以上,线性范围适当,最小检出限均在10pg以下。结论本实验方法可以作为检验生物检材中杀鼠迷、杀鼠灵、溴敌隆和大隆的筛选手段。