液质联用测定人血浆中伪麻黄碱的浓度

目的:建立液质联用法测定人血浆中伪麻黄碱的浓度。方法:采集到的血浆样品以苦参碱为内标,经0.2 mL 0.02 mol·mL~(-1)碳酸钠溶液碱化后氯仿萃取进行 LC-MS 分析。色谱条件为 Kromasil C_(18)柱(150 mm×4.6 mm,5 μm),流动相为乙腈-0.1%甲酸(13:87),流速为1.0 mL·min~(-1);质谱条件为大气压电喷雾离子源(ESI 源),正离子方式检测;扫描方式为选择离子监测(SIM),用于定量分析的离子分别为 m/z 166(伪麻黄碱)和 m/z 249(苦参碱)。结果:方法的线性范围为5～300 ng·mL~(-1),定量下限为5 ng·mL~(-1),提取回收率均大于77.9%,日内、日间精密度(RSD)均小于12.7%。结论:本方法灵敏、专属,适于伪麻黄碱人体血浆药物浓度的测定。     

液相色谱-质谱联用法测定人血浆中多潘立酮

目的:建立测定人血浆中多潘立酮的液相色谱-电喷雾串联质谱(LC/ESI-MS/MS)法。方法:待测血浆0.1 mL 用甲醇沉淀蛋白,取离心后的上清液进样10μL在 Phenomenex Gimim-C_(18)柱(2.00 mm×50 mm,5 μm)上分离,流动相为甲醇-水(40:60,v/v,含0.3%醋酸),流速0.2 mL·min~(-1),LC/ESI-MS/MS 采用多离子反应监测,正离子模式,用于定量分析的离子反应分别为 m/z426→m/z175(多潘立酮)和 m/z 379→m/z 264(氨溴索,内标)。结果:血浆中的内源性物质不干扰测定,每个样品分析时间约2 min;本法线性范围为0.3～100 ng·mL~(-1),最低定量浓度为0.3 ng·mL~(-1);日内、日间 RSD 分别小于7.1%和13.3%,相对误差小于5.4%。结论:该法操作简便、快速、准确,灵敏度高,可用于多潘立酮临床治疗剂量的药物动力学研究。     

Beagle犬血浆中辛伐他汀的LC-MS测定

建立了LC-MS法测定Beagle犬血浆中的辛伐他汀。采用C_(18)柱,流动相为乙腈-0.1%甲酸溶液(60:40),以洛伐他汀为内标,采用电喷雾离子化法(ESI)采集正离子,单离子方式检测m/z 419(辛伐他汀)和m/z 405(洛伐他汀)。辛伐他汀在0.5～160ng/ml浓度范围内线性良好,检测限为0.25ng/ml。血浆样品的日内、日间RSD均小于7.7%,方法回收率为92.2%～99.2%,提取回收率为92.8%～100.9%。     

LC／MS／MS测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度

目的:用高效液相色谱-质谱联用方法同时测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度。方法:大鼠血浆用乙醚萃取法处理后,采用 LC/MS/MS 方法,测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度。HPLC 条件:大连依利特 Hypersil BDS C_(18)柱(2.1 mm×50 mm,3μm),流动相:水(含0.1%甲酸)-乙腈(39:61,v/v),柱温:12℃,流速:200 μL·min~(-1),进样量:10 μL;质谱检测参数为电喷雾电离源(ESI);喷雾电压:3500 V;碰撞压力:0.1333 Pa;源内碰撞诱导电压:普萘洛尔,10 V;4-羟普萘洛尔,15 V;N-去异丙基普萘洛尔,15 V;扫描时间:0.3 s;检测离子:普萘洛尔 m/z 260[M H]~ ;4-羟普萘洛尔 m/z 276[M H]~ ;N-去异丙基普萘洛尔218[M H]~ 。结果:普萘洛尔线性范围2—1000 ng·mL~(-1),最低定量限为2 ng·mL~(-1)。4-羟普萘洛尔和 N-去异丙基普萘洛尔的线性范围1～200 ng·mL~(-1),最低定量限为1 ng·mL~(-1)。普萘洛尔的萃取回收率在90%以上,4-羟普萘洛尔和 N-去异丙基普萘洛尔的萃取回收率均为50%以上,日内、日间 RSD 皆小于15%。结论:适用于测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度及药动学的研究。     

液相色谱-电喷雾质谱联用法测定人血浆中二甲双胍

目的:建立测定人血浆中二甲双胍浓度的液相色谱-电喷雾质谱联用(LC/ESI-MS-MS)法。方法:待测血浆0.2 mL经甲醇沉淀除去蛋白,离心,取上清液5 μL在氰基柱上分离,流动相为甲醇-20 mmol·L~(-1)醋酸铵水溶液(30:70),流速为1.0 mL·min~(-1),LC/ESI-MS-MS 多反应离子检测,正离子模式,用于定量分析的离子分别是二甲双胍 m/z 130.1→70.7和吗啉双胍(内标)m/z 172.2→59.6。结果:血浆中无干扰测定的内源性物质,每个样品分析时间小于5 min,线性范围为10.08～1008 ng·mL~(-1),定量下限为10.08 ng·mL~(-1),日内、日间精密度均小于10%,提取回收率大于85%。结论:该法操作快速、简单、准确、灵敏度高,适用于临床药代动力学研究。     

液相色谱-质谱／质谱联用法测定人血浆中加兰他敏

目的:建立测定人血浆中加兰他敏浓度的液相色谱-质谱/质谱联用法,用于其人体血药浓度测定。方法:血浆样品经液-液萃取后,以甲醇-水-甲酸(65:35:1)为流动相,Zorbax SB-C_8柱(150mm×4.6mm,5μm)分离,采用大气压化学电离源,以选择反应监测方式进行正离子检测。内标为双氢吗啡酮,用于定量分析的离子反应分别为 m/z 288→m/z 213(加兰他敏)和 m/z 286→m/z 185(内标)。结果:血浆中加兰他敏最低定量限为0.5ng·mL~(-1),其线性范围为0.5～100ng·mL~(-1)。其高、中、低3个浓度的平均提取回收率为80.9%,方法回收率为100.5%,日内及日间 RSD 均<8%。结论:方法选择性强,灵敏度高,快速准确,可作为加兰他敏人体内药动学研究的手段。     

LC-MS/MS法测定人血浆中的氨氯地平

目的 采用LC-MS/MS测定人血浆中的氨氯地平.方法 血浆样品经氢氧化钠碱化,乙酸乙酯萃取后进行分析.使用Gmini C18 110A(50 mm×3 mm,5 μm)色谱柱,流动相为甲醇-水-甲酸(55:45:0.025),流速0.2 ml·min-1,通过电喷雾离子化四极串联质谱,以多反应离子方式检测.氨氯地平m/z 409.4→237.9和莫沙比利m/z 422.1→198.0.结果 线性范围为0.0313～20 ng·ml-1,最低定量限为0.0313 ng·ml-1,萃取回收率为74.4%～83.1%,方法回收率为96.1%～108.7%,日内和日间RSD均<8%.结论 所用方法快速、简便、灵敏度高,适用于人血浆中药物浓度的测定及药物动力学和生物利用度的研究.     

UHPLC-MS/MS同时测定人血浆中辛伐他汀及其代谢产物

目的采用UHPLC-MS/MS同时测定人血浆中辛伐他汀及其代谢产物辛伐他汀酸,并研究辛伐他汀、辛伐他汀酸在人体内的药动学特征。方法血浆样品以乙醚萃取,采用UHPLC-MS/MS进行分析。色谱柱:Agilent ZORBAX SB-C18(100mm×2.1mm,3.5μm);乙腈-1mmol·L-1醋酸铵(甲酸调pH 4.5)为流动相梯度洗脱,流速:0.2 m L·min-1。采用电喷雾离子源(ESI),以多反应监测方式(MRM)进行定量分析。辛伐他汀和内标洛伐他汀在正离子模式下定量分析,离子对分别为m/z 419.4→199.3和m/z 405.3→199.3;辛伐他汀酸和内标洛伐他汀酸在负离子模式下定量分析,离子对分别为m/z435.5→115.2和m/z 421.4→101.2。结果辛伐他汀和辛伐他汀酸的线性范围均为0.2~50 ng·m L-1(r>0.99),最低定量限均为0.2 ng·m L-1,日内和日间精密度(RSD)均≤11.10%,提取回收率均≥63.71%。结论该方法专属性强、灵敏度高、重现性好,适用于辛伐他汀的药动学研究。     

液相色谱-质谱联用法测定大鼠血浆中盐酸金刚烷胺浓度及体内药动学研究

目的:建立测定大鼠血浆中盐酸金刚烷胺的液相色谱-质谱联用法。方法:色谱柱为 Agilent Zorbax SB-C_(18)(2.1 mm×150 mm,3.5μm);乙腈-0.05%甲酸溶液(28∶72)为流动相,流速0.2 mL·min~(-1);柱温25℃;内标为盐酸美金刚。质谱条件为电喷雾电离源(ESI),选择正离子抽提方式检测,金刚烷胺和美金刚的选择检测离子分别为 m/z 152.20([M H]~ )、m/z180.20([M H]~ )。选用 SD 大鼠6只,单剂量尾静脉注射给予1 mg·kg~(-1),进行了血药浓度的测定。结果:盐酸金刚烷胺血浆线性范围为0.9824～491.2 ng·mL~(-1),检测限为0.025 ng·mL~(-1),方法回收率均大于95%,日内、日间精密度试验的 RSD<5.0%。结论:本方法专属性强,灵敏度高,线性关系良好,操作简便,适用于药代动力学研究。     

HPLC-MS／ESI同时测定人血浆中文拉法新及其3种代谢产物

目的:建立一种快速灵敏的反相高效液相色谱-质谱联用法同时测定人血浆中文拉法新(VEN)和其3种代谢产物:氧云甲基文拉法新(ODV)、氮去甲基文托法新(NDV)和氮、氰双去甲基文拉法新(DDV)的浓度。方法:以艾司唑仑为内标,样本碱化后用乙醚捉取2次,合并有机相,40℃氮气吹干,100μL流动相定容;用 BDS HYPERSIL C_(18)反相色谱柱(250mm×4.6mm,5μm)进行分离,以乙腈-缓冲溶液(30 mmol·L~(-1)醋酸铵 2.6mmol·L~(-1)甲酸 0.13 mmol·L~(-1)三氟乙酸)(40:60,v/v)为流动相,采用梯度洗脱程序,柱温为50℃,流速为1.0 mL·min~(-1)。采用质谱电喷雾电离源(ESI)将样品离子化,选择性离子监测(SIM)准分子离子和/或主要碎片离子峰。结果:VEN、ODV、NDV、DDV 以及内标艾司唑仑在6min 内完全分离;各物质在2～900 ng·mL~(-1)时线性关系良好,相关系数均大于0.9991;萃取回收率均大于77%;方法回收率均大于91%;最低检测浓度:VEN 0.4 ng·mL~(-1)、ODV0.2 ng·mL~(-1)、NDV 0.3 ng·mL~(-1)、DDV 0.2 ng·mL~(-1);日内日间RSD 均小于11%。结论:本方法简单快速,灵敏准确,可用于药物动力学、药物代谢机制以及血药浓度的临床监护、中毒分析的研究。     

API-ionspray MS and MS/MS study on the structural characterization of bisbenzylisoquinoline alkaloids

API-ionspray MS and MS/MS techniques have been utilized to elucidate the structures of 20 bisbenzylisoquinoline alkaloids, consisting of 17 diether and three monoether links of two benzyltetrahydroisoquinoline units, which were isolated and identified previously from a variety of Thalictrum sp. (Ranunculaceae family). Apparent protonated molecular ions ([M+H](+)) and very intense doubly-protonated molecular ion ([M+2H](++), 100% of relative abundance) in Q1 Scan MS spectra and prominent as well as diagnostic product ions for the structural information in MS/MS spectra were observed in nanogram quantities for all investigated alkaloids.     

LC/MS/MS的多反应监测方法定量测定灯盏乙素

目的：建立一种可靠的灯盏乙素定量分析方法。方法：用三级四极串联质谱(MS/MS)作为HPLC的检测器,其中MS/MS使用了多反应监测(MRM)扫描方式。选择母→子离子对m/z -461→m/z -285作为MRM监测的离子对;HPLC流动相为100％甲醇,流速0.9 mL.min^-1,色谱柱Beckman ODS-1。以测定短葶飞蓬提取物的灯盏乙素含量为例,对此方法进行了应用。结果：灯盏乙素在短葶飞蓬提取物中含量为6.98%。方法线性范围20～160 ng.mL^-1 (γ=0.999);加入灯盏乙素标准品20,60和160 ng的加样回收率分别为：96.5%,97.4%和97.3%。检测限为1 ng,每个样品的分析时间为4 min。结论：此法灵敏、快速、准确,可应用于灯盏乙素的各种药剂、药代的研究。     

System suitability in bioanalytical LC/MS/MS

System suitability is widely recognized as a critical component of bioanalysis. This paper discusses a generic system suitability test that monitors instrument performance throughout a run when used for liquid chromatography tandem mass spectrometry (LC/MS/MS) in bioanalysis. This system suitability process is designed to ensure that the LC/MS/MS system is performing in a manner that leads to the production of accurate and reproducible data that can be submitted with confidence to regulatory agencies. This process contains tests for signal stability, carryover, and instrument response. This approach is integrated throughout an analytical run and has been used in the analysis of over 25,000 batches of clinical samples. Two case studies are presented in which quality control samples and standards meet all acceptance criteria (based on Standard Operating Procedures and the Food and Drug Administration's recommendations for bioanalytical method validation) but failed the proposed system suitability test, and thus were rejected. In these case studies, the concentrations of a significant number of clinical samples (over 35%) were affected, resulting in changes of more than 15% when the samples were reanalyzed. These data indicate that the poor performance of an LC/MS/MS system could adversely affect the calculated concentrations of unknown samples even though the results for quality control samples appear to be acceptable.     

LC/MS/MS与GC/MS法分析鉴定小鼠尿中沙美特罗的代谢产物

目的:鉴定沙美特罗在小鼠尿中的主要代谢产物.方法:ig给药后,收集小鼠尿液,经固相提取,葡萄糖醛酸酶水解,进行LC/MS/MS分析和硅烷化后进行GC/MS分析同时分离鉴定沙美特罗代谢产物.结果和结论:在给药后尿样中发现沙美特罗原型和4种代谢产物M1～M4,其结构推测为19-羟基沙美特罗(M1)、2-羰基沙美特罗(M2)、19-羰基沙美特罗(M3)和19-羟基-8-甲氧基沙美特罗(M4).     

液相色谱-串联质谱法快速测定人血浆中伊曲康唑

目的:建立灵敏、快速的液相色谱-串联质谱法测定人血浆中伊曲康唑,并用于药代动力学研究。方法:血浆样品经乙腈沉淀蛋白后,以乙腈-水-甲酸(80:20:0.2,v/v/v)为流动相,流速0.50 mL·min~(-1),Zorbax sB C_(18)柱分离,采用电喷雾电离源,以选择反应监测(SRM)方式进行正离子检测。用于定量分析的离子反应分别为 m/z 705→m/z(392 432)(伊曲康唑)和m/z 256→m/z 167(内标,苯海拉明)。结果:测定血浆中伊曲康唑的线性范围为1.00～1000 ng·mL~(-1),定量下限为1.00 ng·mL~(-1)。日内、日间精密度(RSD)均小于7.2%,准确度(RE)在±3.0%以内。应用本法测得20名健康受试者单剂量口服200mg 伊曲康唑胶囊后的主要药动学参数为:T_(max)(4.25±1.02)h,C_(max)(155.5±73.3)ng·mL~(-1),t_(1/2)(20.4±11.4)h;用梯形法计算,AUC_(0-84h)(2257±1168)ng·h·mL~(-1),AUC_(0-∞)(237.1±1207)ng·h·mL~(-1)。结论:该法选择性强、灵敏度高、操作简便,适用于伊曲康唑的临床药代动力学研究。     

Analysis of 'SHENMAI' injection by HPLC/MS/MS

An HPLC/MS/MS method was developed for the analysis of 'SHENMAI' injection, composed of red ginseng and ophiopogon. The constituents of 'SHENMAI' were found to be similar with those of ginseng and 39 ginsenosides were detected. By the studies of MS and MS/MS spectra and the comparison with literature data, most of these ginsenosides were identified. Based on this study, suggestions were put forward to improve the quality control system of 'SHENMAI' injection.     

超高压液相/电喷雾-LTQ-Orbitrap质谱联用技术分析三七根中皂苷类成分

建立三七药材的UPLC-ESI-HRMS定性分析方法。采用Kinetics色谱柱,梯度洗脱,经DAD检测器后采用ESI-LTQ-Orbitrap质谱负离子模式采集,三七成分的一级全扫描离子在离子阱内进行CID多级质谱,FT检测高分辨质谱数据进行分析。结果表明三七皂苷成分分离良好,根据多级质谱碎片信息和精确相对分子质量信息,结合文献,从三七中鉴定分析了43个成分,并首次从三七中检测到新型的三七皂苷母核和乙酰取代皂苷成分。本方法分离度良好、灵敏度高,可用于三七药材的定性分析,并有助于对三七进行进一步的活性成分研究和质量控制。     

Determination of a selection of synthetic cannabinoids and metabolites in urine by UHPSFC‐MS/MS and by UHPLC‐MS/MS

Two different analytical techniques, ultra‐high performance supercritical fluid chromatography‐tandem mass spectrometry (UHPSFC‐MS/MS) and reversed phase ultra‐high performance liquid chromatography‐tandem mass spectrometry (UHPLC‐MS/MS), were used for the determination of two synthetic cannabinoids and eleven metabolites in urine; AM‐2201 N‐4‐OH‐pentyl, AM‐2233, JWH‐018 N‐5‐OH‐pentyl, JWH‐018 N‐pentanoic acid, JWH‐073 N‐4‐OH‐butyl, JWH‐073 N‐butanoic acid, JWH‐122 N‐5‐OH‐pentyl, MAM‐2201, MAM‐2201 N‐4‐OH‐pentyl, RCS‐4 N‐5‐OH‐pentyl, UR‐144 degradant N‐pentanoic acid, UR‐144 N‐4‐OH‐pentyl, and UR‐144 N‐pentanoic acid. Sample preparation included a liquid‐liquid extraction after deconjugation with ß‐glucuronidase. The UHPSFC‐MS/MS method used an Acquity UPC^{2 TM} BEH column with a mobile phase consisting of CO₂ and 0.3% ammonia in methanol, while the UHPLC‐MS/MS method used an Acquity UPLC® BEH C₁₈ column with a mobile phase consisting of 5 mM ammonium formate (pH 10.2) and methanol. MS/MS detection was performed with positive electrospray ionization and two multiple reaction monitoring transitions. Deuterated internal standards were used for six of the compounds. Limits of quantification (LOQs) were between 0.04 and 0.4 µg/L. Between‐day relative standard deviations at concentrations ≥ LOQ were ≤20%, with biases within ±19%. Recoveries ranged from 40 to 90%. Corrected matrix effects were within 100 ± 10%, except for MAM‐2201 with UHPSFC‐MS/MS, and for UR‐144 N‐pentanoic acid and MAM‐2201 N‐4‐OH‐pentyl with UHPLC‐MS/MS. Elution order obtained by UHPSFC‐MS/MS was almost opposite to that obtained by UHPLC‐MS/MS, making this instrument setup an interesting combination for screening and confirmation analyses in forensic cases. The UHPLC‐MS/MS method has, since August 2014, been successfully used for confirmation of synthetic cannabinoids in urine samples revealing a positive immunoassay screening result. Copyright © 2015 John Wiley & Sons, Ltd.     

LC／MS／MS技术对西他沙星代谢物结构的研究

利用LC／MS／MS实现对西他沙星体内代谢物的在线鉴定,并通过定向合成的代谢物对照品,对西他沙星代谢物的结构作了进一步验证。结果鉴定出六个代谢物D2～D7。根据代谢物的结构特征证明了西他沙星在肝脏经氧化被代谢消除。