气相色谱-串联质谱法同时检测尿液中15种有毒生物碱

在气相色谱-串联质谱(GC-MS/MS)多反应监测(MRM)模式下,建立了对尿液中八角枫碱、芦竹碱、毒扁豆碱、毛果芸香碱、哈尔碱、氧化苦参碱、黄华碱、钩吻素子、钩吻碱、延胡索乙素、吴茱萸碱、血根碱、白屈菜红碱、士的宁和马钱子碱15种有毒生物碱的定性定量分析。对样品前处理、色谱、质谱条件进行了优化。在优化条件下,毒扁豆碱、哈尔碱、钩吻素子和士的宁在20~800μg/L范围内线性关系良好,其余生物碱在40~800μg/L范围内线性关系良好,相关系数均不小于0.993 2。在高、中、低3种加标水平下,除八角枫碱的平均回收率为60.0%~68.3%外,其余14种生物碱的平均回收率为81.9%~114.4%,各生物碱的相对标准偏差(RSD)不大于17.6%。方法的检出限(LOD)为4~20μg/L,定量下限(LOQ)为10~40μg/L。该方法操作简便、快捷、灵敏,适用于中毒患者尿液中有毒生物碱成分的检测。     

液相色谱-电喷雾串联质谱同时检测血液中8种有毒生物碱

建立了高效液相色谱-电喷雾串联质谱同时检测血液中8种有毒生物碱的方法.血液样品经过乙酸铵-氨水缓冲溶液(pH≈9)处理后,以甲醇提取,采用电喷雾电离(ESI)、多反应监测(MRM)方式,可同时对麻黄碱、毛果芸香碱、士的宁、阿托品、钩吻素子、马钱子碱、乌头碱、喜树碱8种有毒生物碱进行定性和定量分析.在优化的条件下,上述前4种组分在1～100 μg/L范围内线性关系良好,后4种组分在2.5～500 μg/L范围内线性关系良好.8种有毒生物碱的提取回收率在83.1%～104.0%范围内.士的宁、阿托品、马钱子碱、乌头碱和钩吻素子检出限为0.05 μg/L,定量限为0.1 μg/L;麻黄碱、毛果芸香碱、喜树碱的检出限为0.1 μg/L,定量限为0.5 μg/L.各组分的日内RSD小于9%,日间RSD小于10%.本方法操作简便快捷、选择性好、灵敏度高,适用于中毒诊断和法医毒物分析.     

亲水液相色谱三重四极杆质谱联用法快速检测尿液和血浆中河豚毒素

建立了快速检测尿液和血浆中河豚毒素的亲水液相色谱三重四极杆质谱联用分析方法.样品经乙腈沉淀、TSK-gel Amide-80亲水色谱柱分离后,采用电喷雾串联质谱多反应监测(MRM)模式检测,基体匹配标准外标法定量.尿液和血浆样品的线性范围分别为3～500 μg/L和1～200 μg/L,加标回收率分别为96%～108%和100%～105%,相对标准偏差小于9%和16%(n=6),样品的检出限分别为1.0和0.3 μg/L(S/N=3).本方法简单、快速、灵敏、特异性强.     

超高效液相色谱三重四极杆质谱法同时快速测定血浆和尿液中11种杀鼠剂

建立了血浆和尿液中11种杀鼠剂同时测定的超高效液相色谱串联质谱方法.血浆样品经乙腈沉淀、尿液样品经乙酸乙酯萃取,氮吹浓缩后用初始流动相复溶,在UPLC BEH C18柱上以甲醇-4 mmol/L乙酸铵为流动相进行梯度洗脱分离,负离子电喷雾多反应监测模式检测,基质标准外标法定量分析.血浆和尿液中11种杀鼠剂的加标回收率分别在68%～118%和62%～104%之间;日内相对标准偏差分别为5.6%～21%和4.1%～18%;日间相对标准偏差分别为8.5%～26%和7.1%～ 25% (n=6).除尿液中安妥的定量限为5 μg/L外,其余待测物的定量限均为1 μg/L.本方法灵敏度高,操作简单,适于中毒病人的快速诊断检测.     

超高效液相色谱-串联质谱法同时测定尿样中4种痕量的乌头类生物碱

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)结合中空纤维微萃取(HF-LPME)同时检测尿样中痕量的乌头碱、次乌头碱、新乌头碱和滇乌头碱等4种生物碱的方法。采用HF-LPME对尿样进行提取、纯化和富集,富集倍数达102～301。同时采用电喷雾电离(ESI)、多反应监测(MRM)进行含量测定,显著提高了尿液中乌头类生物碱的检测灵敏度,4种乌头类生物碱的定量限达到0.01～0.1 ng/L,可大大延长中毒患者尿样中乌头类生物碱的检测时间窗。方法验证结果表明,尿液中乌头碱、新乌头碱和滇乌头碱在0.01～10 ng/L、次乌头碱在0.1～100 ng/L范围内线性关系良好,提取回收率为80.2%～109%,相对标准偏差小于4.6%。该方法适用于乌头类生物碱中毒案件的检测,为痕量乌头类生物碱分析提供了灵敏的分析方法。     

高效液相-电喷雾串联质谱法测定动物肌肉组织中的糖皮质激素

本实验建立了肌肉组织中氢化可的松、可的松、泼尼松和地塞米松含量的高效液相色谱-电喷雾串联质谱(LC-ESI-MS/MS)检测方法.样品经酶水解、乙酸乙酯提取、HLB固相萃取净化,C8色谱柱分离,在多反应选择性监测模式(MRM)下采用负离子模式进行信号采集和测定.4种糖皮质激素的检出限为0.13 ～0.25 μg/kg,定量限为0.25 ～ 0.5 μg/kg.在0.5～50.0 μg/L范围内具有良好的线性关系(R2＞0.99).在0.5,5.0和10.0μg/kg基质加标水平下,4种物质的平均回收率为74.0％ ～ 101.8％,相对标准偏差0.7％ ～8.6％.     

蔬菜中26种农药残留的高效液相色谱-串联质谱法测定

建立了蔬菜中26种农药的反相高效液相色谱-电喷雾串联质谱 (LC-ESI MS/MS) 检测方法.蔬菜样品经含0.1%醋酸的乙腈提取、浓缩、分散型固相萃取净化,液相色谱串联质谱测定,内标法定量.26种农药在5.0 ～200 μg/L范围内线性关系良好(r2＞0.997).在豌豆、菠菜、脱水葱3种基质中的检出限和定量下限均为2.0 μg/kg和5.0 μg/kg.在5.0、10.0、40.0 μg/kg 3个添加水平下,26种农药的回收率为70% ～110%,精密度(RSD)小于15%.方法准确、灵敏、简单,适用于蔬菜中26种农药残留的同时测定.     

Determination of Sulbactam in Milk by Hydrophilic Interaction Chromatography Tandem Mass Spectrometry ( HILIC - MS/MS)

建立了亲水作用色谱-串联四极杆质谱测定液态奶中微量舒巴坦的分析方法.样品经0.2％乙酸水溶液提取,HLB固相萃取柱净化、富集,以甲酸铵-乙腈为流动相,经Acquity UPLC BEH HILIC色谱柱分离后采用电喷雾质谱多反应监测方式(MRM)扫描,外标法定量.结果表明,舒巴坦的质量浓度在1～100 μg/L范围内线性关系良好,r2大于0.99,定量下限为1.0 μg/kg.加标水平在1.0～50.0 μg/kg范围时,回收率为82％～102％,相对标准偏差(RSD)为1.6％ ～4.7％.该方法前处理简便快捷、灵敏度高、回收率和重现性良好,适用于液态奶中舒巴坦的测定.     

固相萃取-气相色谱-串联质谱法同时测定人体尿液中4种有机磷农药代谢物

建立了同时测定尿液中4种有机磷类农药代谢物的气相色谱/串联质谱分析方法.尿液样品以WCX固相萃取柱富集提取,乙酸乙酯-乙腈(70∶30,V/V)再萃取,浓缩干燥,甲苯溶解,N-叔丁基二甲基甲硅烷基-N-甲基三氟乙酰胺衍生化.采用HP-5MS色谱柱(30 m×0.25 mm×0.25 μm)程序升温分离,串联质谱多反应检测,内标法定量.通过比较尿样中代谢物在不同萃取溶剂、固相萃取柱及pH值洗脱液等条件下的回收率,优化了前处理方法.在0.2～ 200 μg/L范围内,4种代谢物峰面积与内标峰面积的比值与质量浓度的线性关系良好(R2≥0.992),方法检出限为0.083 ～0.667μg/L,定量限为0.2～2.0 μg/L,平均回收率为54.1％～68.6％,相对标准偏差均小于8.5％(n=6).本方法稳定、可靠,分析时间短,不需要使用大量有机溶剂,可同时处理大批量样品,适用于人群有机磷类农药暴露情况的评估.     

液相色谱串联质谱法快速测定猪尿中10种镇静剂类药物残留量

建立了快速测定猪尿中10种镇静剂类药物(噻拉嗪、阿扎哌隆、氟哌啶、氟哌啶醇、艾司唑仑、硝西泮、地西泮、奥沙西泮、氯丙嗪和奋乃静)残留量的液相色谱串联质谱方法.猪尿样品离心后过C18固相萃取小柱,用甲醇-乙酸乙酯(1∶ 4, V/V)混合溶剂洗脱,氮气吹干后用0.1%甲酸溶液溶解进行仪器分析.采用Eclipse XDB-C18色谱柱分离,以乙腈和0.1%甲酸溶液为流动相进行梯度洗脱,电喷雾正电子(ESI+)模式电离,多反应监测(MRM)模式检测,基质校准进行定量分析.10种镇静剂类药物在5～200 μg/L范围内呈良好的线性,线性相关系数均大于0.99.方法检出限为0.11～0.52 μg/L; 定量限为0.20～0.91 μg/L.添加浓度为1.0, 2.0和10.0 μg/L时,平均回收率在74.6%～115 %之间,批内和批间相对标准偏差均小于15%.     

丹磺酰氯柱前衍生-超高效液相色谱-串联质谱法测定人体尿样中的环己胺

建立丹磺酰氯柱前衍生-超高效液相色谱-串联质谱法测定人体尿样中环己胺的方法。冷冻样品经解冻、离心后,用丹磺酰氯衍生,固相萃取小柱净化。目标化合物采用 Waters ACQUITY CSHTM C18色谱柱(50 mm×2.1 mm,1.7μm)分离,以甲醇和0.002 mol/L乙酸铵溶液为流动相梯度洗脱,采用电喷雾离子源电离、正离子多反应监测模式质谱检测。环己胺在2.5~200μg/L浓度范围内有较好的线性关系,相关系数大于0.999,回收率为98.7%~102.3%,精密度为3.1%~5.2%,检出限和定量限分别为1.0和3.0μg/L。结果表明,本方法操作简单、准确可靠,可适用于人体尿液中环己胺的定量分析。应用本方法测定200份学生尿液样品,环己胺检出率为34.5%。     

Capillary electrophoresis with LED-induced native fluorescence detection for determination of isoquinoline alkaloids and their cytotoxicity in extracts of Chelidonium majus L

In this study, we introduced a simple and sensitive method of capillary electrophoresis with ultraviolet light-emitting diode-induced native fluorescence (UV-LEDIF) detection for the determination of isoquinoline alkaloids in extracts of Chelidonium majus L. Samples were extracted with acidic methanol and the extracts were directly analysed by CE. Simultaneous determination of protopine, chelidonine, coptisine, sanguinarine, allocryptopine, chelerythrine and stylopine was performed in 20mM phosphate buffer (pH 3.1). The baseline separation of these alkaloids was finished within 20 min. As these alkaloids have native fluorescence, they were directly detected using the commercially available UV light emitting diode without troublesome fluorescent derivatisation. Satisfactory LOD values were obtained for the studied compounds considering their appearance in natural extracts. Lower limits of detection were 0.05 μg/mL for protopine, 0.06 μg/mL for stylopine and allocryptopine, 0.07 μg/mL for chelidonine, 0.22 μg/mL for sanguinarine, 1.7 μg/mL for chelerythrine and 5.5 μg/mL for coptisine. The developed method was successfully applied to determine the contents of seven alkaloids in the aerial parts of Chelidonium majus L, which varied from 0.025 to 0.763% (w/w). Also, to demonstrate the potential of the proposed CE method, an estimation of the cytotoxic properties of selected Celandine alkaloids in a natural extract was carried out.     

Alkaloids induce programmed cell death in bloodstream forms of trypanosomes (Trypanosoma b. brucei)

The potential induction of a programmed cell death (PCD) in Trypanosoma b. brucei by 55 alkaloids of the quinoline, quinolizidine, isoquinoline, indole, terpene, tropane, steroid, and piperidine type was studied by measuring DNA fragmentation and changes in mitochondrial membrane potential. For comparison, the induction of apoptosis by the same alkaloids in human leukemia cells (Jurkat APO-S) was tested. Several alkaloids of the isoquinoline, quinoline, indole and steroidal type (berberine, chelerythrine, emetine, sanguinarine, quinine, ajmalicine, ergotamine, harmine, vinblastine, vincristine, colchicine, chaconine, demissidine and veratridine) induced programmed cell death, whereas quinolizidine, tropane, terpene and piperidine alkaloids were mostly inactive. Effective PCD induction (EC(50) below 10 microM) was caused in T. brucei by chelerythrine, emetine, sanguinarine, and chaconine. The active alkaloids can be characterized by their general property to inhibit protein biosynthesis, to intercalate DNA, to disturb membrane fluidity or to inhibit microtubule formation.     

Simultaneous determination of six toxic alkaloids in human plasma and urine using capillary zone electrophoresis coupled to time‐of‐flight mass spectrometry

A novel capillary zone electrophoresis separation coupled to electro spray ionization time‐of‐flight mass spectrometry method was developed for the simultaneous analysis of six toxic alkaloids: brucine, strychnine, atropine sulfate, anisodamine hydrobromide, scopolamine hydrobromide and anisodine hydrobromide in human plasma and urine. To obtain optimal sensitivity, a solid‐phase extraction method using Oasis MCX cartridges (1 mL, 30 mg; Waters, USA) for the pretreatment of samples was used. All compounds were separated by capillary zone electrophoresis at 25 kV within 12 min in an uncoated fused‐silica capillary of 75 μm id × 100 cm and were detected by time‐of‐flight mass spectrometry. This method was validated with regard to precision, accuracy, sensitivity, linear range, limit of detection (LOD), and limit of quantification (LOQ). In the plasma and urine samples, the linear calibration curves were obtained over the range of 0.50–100 ng/mL. The LOD and LOQ were 0.2–0.5 ng/mL and 0.5–1.0 ng/mL, respectively. The intra‐ and interday precision was better than 12% and 13%, respectively. Electrophoretic peaks could be identified by mass analysis.     

高效液相色谱-离子阱质谱法测定生物体液中的痕量敌鼠和氯敌鼠

建立了中毒应急检测样品生物体液内痕量敌鼠和氯敌鼠的高效液相色谱-离子阱质谱定性定量检测方法。全血样品由甲醇/乙腈(50/50, v/v)沉淀蛋白后再经Oasis HLB固相萃取小柱净化富集,尿液样品直接经同类小柱净化富集。采用Extend C18柱分离,以乙酸铵-乙酸(0.02 mol/L, pH 5.5)缓冲溶液和甲醇(15/85, v/v)溶液为流动相,使用电喷雾电离负离子多反应监测模式检测敌鼠和氯敌鼠。对全血样品,敌鼠和氯敌鼠分别在1.0～200.0 μg/L和0.5～100.0 μg/L范围内呈良好的线性关系,回收率分别在90.1%～92.2%和87.6%～93.4%范围内,日内相对标准偏差(RSD)分别小于6.8%和7.1%,日间RSD分别小于9.9%和10.9%,定量限分别为1.0 μg/L和0.5 μg/L。对尿液样品,敌鼠和氯敌鼠分别在0.2～ 40.0 μg/L和0.1～20.0 μg/L范围内呈良好的线性关系,回收率分别在90.1%～94.5%和90.0%～ 98.0%范围内,日内RSD分别小于6.1%和7.3%,日间RSD分别小于8.9%和11.2%,定量限分别为0.2 μg/L和0.1 μg/L。本方法简便、灵敏,能满足敌鼠和氯敌鼠中毒病人的临床快速诊断要求。     

高效液相色谱法检测胃液中色氨酸和利多卡因

采用高效液相色谱-紫外-荧光和高效液相色谱-紫外-质谱联用法,证明胃癌患者胃液中存在的荧光物质主要为色氨酸.建立了反相高效液相色谱-双波长同时紫外检测法定量分析胃液中色氨酸和利多卡因的方法.使用Kromasil C18柱(150 mm×4.6 mm, 5 μm),以含0.1%三氟乙酸的甲醇溶液与含0.1%三氟乙酸水溶液为流动相,梯度洗脱分析.色氨酸使用278 nm检测,浓度在0.5～200 mg/L范围内峰面积对浓度的线性相关系数为0.9994;利多卡因使用254 nm检测,浓度在20～5000 mg/L范围内相关系数为0.9992.色氨酸和利多卡因的检出限分别为0.15和5 mg/L;平均加标回收率分别为70.8%～110.4%(色氨酸) 和87.1%～116.2% (利多卡因).利用本方法对38例胃癌患者和48例非胃癌患者的胃液进行测试,两组胃液中色氨酸的含量存在一定的差异.     

Solidification of floating organic droplet microextraction for determination of seven insecticides in fruit juice,vegetables and agricultural runoff using gas chromatography with flame ionization and mass spectrometry detection

Liquid microextraction employing solidification of the floating organic droplet, with vortexing and heating to optimize extraction efficiency, was developed for the determination of seven insecticides in fruit juice, vegetables, and agricultural runoff water. The extracts were analyzed by gas chromatography with both flame ionization and mass spectrometry detection for the determination of chlorpyrifos, prothiofos, profenofos, ethion, λ‐cyhalothrin, permethrin, and cypermethrin, respectively. Using 20 μL of 1‐undecanol in 10 mL of aqueous solution containing 1% w/v sodium chloride provided preconcentration factor of 500. The enrichment factor of the analytes was in the range of 355 to 509 with extraction recovery >71%. The linearity ranges were 4–200 μg/kg for gas chromatography with flame ionization detection and 1–100 μg/kg for gas chromatography with mass spectrometry, with limits of detection ranging from 0.04 to 1.2 μg/kg, which are lower than the international maximum residue limits for vegetables and fruit juice. Intra‐day and inter‐day precisions are less than 5.4 and 7.0% relative standard deviation, respectively. The method was successfully applied to the determination of the seven insecticides in samples of vegetables, fruit juice and agricultural runoff, with recoveries ranging from 61.7 to 120.8%. The extraction method is simple, efficient and environmentally friendly.     

Simultaneous determination of atropine,scopolamine, and anisodamine in Flos daturae by capillary electrophoresis using a capillary coated by graphene oxide

A novel CE method was developed for the separation and determination of three main tropane alkaloids in Flos daturae with a capillary coated by graphene oxide (GO). The GO‐coated capillary was characterized by SEM, energy dispersive X‐ray spectroscopy, and Raman spectroscopy, and the results indicated that the inner surface of the capillary was partially coated by GO. A phosphate solution (40 mM, pH7.0) containing 20% v/v methanol and 30% v/v acetonitrile was used as the running buffer for the analysis of the atropine, scopolamine, and anisodamine. The linear ranges of atropine, scopolamine, and anisodamine was 0.5–200 μg/mL with satisfactory correlation coefficients (R²) > 0.9987, and this novel method provided an efficient separation for three tropane alkaloids as well as a good reproducibility and stability. Finally, the method was successfully applied for the determination of these three tropane alkaloids in plant extracts.     

自制混合型小柱净化-高效液相色谱-串联质谱法同时测定尿液中有机磷酸酯代谢物和8-羟基-2'-脱氧鸟苷

建立了基于自制混合型小柱的样品净化-高效液相色谱-串联质谱同时测定7种有机磷酸酯（OPEs）主要代谢产物及生物标志物8-羟基-2'-脱氧鸟苷（8-OHdG）的分析方法。样品经乙腈提取后用自制小柱富集净化,以乙腈-0.2%（v/v）氨水溶液作为流动相进行梯度洗脱,在多反应监测模式下进行定性和定量分析。结果显示,8种目标物在0.1~200 μg/L范围内呈良好的线性关系,7种OPEs代谢物的回收率为52.36%~114.56%,8-OHdG回收率为88.63%~97.72%。将该方法应用于人体尿液实际样品中,7种OPEs代谢物和8-OHdG的检出范围分别为6.24~46.07 μg/L和5.90~16.71 μg/L,8-OHdG与7种OPEs代谢物总含量之间存在显著相关性。该方法操作简单、灵敏度高、准确性好、重现性强,可为更全面地评价人体内OPEs暴露水平及机体损伤提供可靠的技术支持。     

高效液相色谱-电喷雾离子阱质谱联用测定尿中的8-羟基脱氧鸟嘌呤

建立了尿样中8-羟基脱氧鸟嘌呤的HPLC-MS测定方法.尿样中的8-羟基脱氧鸟嘌呤采用WCX固相萃取小柱预富集后,以0.5%甲酸-甲醇洗脱,吹干后用0.5 mL流动相溶解剩余物上机测定.采用分子的二级碎片,方法在5.0～500.0 μg/L范围内呈良好线性关系,相关系数r=0.999 4,检出限(S/N=3)为0.50...