HPLC法同时测定注射用艾迪（冻干）中人参皂苷Rg1和人参皂苷Re的含量

目的:建立注射用艾迪(冻干)中人参皂苷Rg_1和人参皂苷Re的含量测定方法。方法:色谱柱为Hypersil C_(18)柱(250mm×4.6mm,5μm),流动相为乙腈-0.05%磷酸溶液(19∶81),流速为1.0mL·min~(-1),检测波长为203nm。结果:人参皂苷Rg_1浓度在20～200μg·mL~(-1),人参皂苷Re浓度在10～100μg·mL~(-1)范围内与峰面积呈良好的线性关系,相关系数分别为0.9997和0.9996;平均回收率分别为100.3%和97.6%(n=9)。结论:本法快速,准确,重复性好,可作为注射用艾迪(冻干)的质量控制方法。     

HPLC-MS/MS法测定10批参须中人参皂苷Rg_1、Re和Rb_1的含量

目的建立同时检测参须中人参皂苷Rg1、Re和Rb1含量的HPLC-MS/MS方法,测定10批参须中人参皂苷Rg1、Re和Rb1的含量。方法采用液质联用(HPLC-MS/MS)法,色谱柱为SHIMADZU VP-ODS(4.6 mm×150 mm,5μm),流动相为乙腈-0.1%甲酸水,梯度洗脱(0~3 min,60%乙腈;3~5 min,90%乙腈),流速0.5 m L·min-1,柱温40℃。质谱条件:正离子检测模式,用于定量分析的离子对分别为:人参皂苷Rg1 m/z 823.3→643.6,人参皂苷Re m/z 969.7→789.6和人参皂苷Rb1 m/z1131.5→365.3。结果人参皂苷Rg1在0.43~27.6μg·m L~(-1)与峰面积线性关系良好,r=0.9950,平均回收率为94.0%(RSD=1.5%);人参皂苷Re在0.46~29.6μg·m L~(-1)与峰面积线性关系良好,r=0.9990,平均回收率为95.3%(RSD=1.1%);人参皂苷Rb1在0.41~26.4μg·m L~(-1)与峰面积线性关系良好,r=0.9980,平均回收率为93.4%(RSD=1.2%)。结论该方法简便、准确、快速,可用于参须中人参皂苷Rg1、Re和Rb1的含量检测。     

RP-HPLC法同时测定西洋参含片中4种成分

目的建立同时测定西洋参含片中人参皂苷Rg_1、人参皂苷Re、人参皂苷Rb_1及拟人参皂苷F11含量的RP-HPLC方法。方法色谱柱为Agilent TC-C18(250 mm×4.6 mm,5μm);流动相为0.5 mL·L~(-1)磷酸溶液(A)-乙腈(B),梯度洗脱;流速:1.0mL·min~(-1);检测波长:203nm;柱温:30℃。结果人参皂苷Rg_1和人参皂苷Re及人参皂苷Rb_1在2.5~500μg·mL~(-1)、拟人参皂苷F11在5.0~1 000μg·mL~(-1)范围内线性关系良好(r≥0.999 4);平均加样回收率均大于95.0%。结论该方法简便、准确,重复性好,可作为西洋参含片的质量控制方法。     

LC-MS／MS法测定人血浆中米格列醇

目的:建立测定人血浆中米格列醇的 LC-MS/MS 法。方法:以盐酸格拉司琼为内标,以 Kromasil CN 柱(2.1 mm×150mm,3.0μm)为分析柱;采用0.05%三氟乙酸乙腈溶液-0.05%三氟乙酸水溶液(80:20)为流动相;流速0.2 mL·min~(-1);柱温45℃。质谱条件为电喷雾电离源(ESI~ ),以选择反应离子监测(SRM)方式进行检测,用于定量分析的反应离子分别为 m/z208.1→146.1(米格列醇),m/z 313.1→138.0(盐酸格拉司琼)。结果:米格列醇在0.01～4.0μg·mL~(-1)浓度范围内线性良好;最低检测限为2.0 ng·mL~(-1)(S/N=4);日内、日间精密度(RSD)均小于7%;低、中、高3个浓度的方法回收率均大于90%。结论:本法专属性强,灵敏度高,样品处理简单,可用于米格列醇临床药物动力学研究。     

HPLC—MS／MS法测定人血浆中赖诺普利的浓度

目的:建立测定人血浆中赖诺普利浓度的 HPLC-MS/MS 法。方法:200μL待测血浆中定量加入内标后,直接加入蛋白沉淀剂三氟醋酸,涡旋离心后取上清液直接进样。采用 Zorbax Eclipse DB-C_8柱(5μm,150 mm×4.6 mm)分析柱;流动相为甲醇-12.5 mmol·L~(-1)醋酸铵缓冲液(60:40,pH=4.40),流速0.5 mL·min~(-1);进样量25μL。质谱检测采用 ESI 正离子模式,扫描方式为多反应监测方式,扫描离子对为 m/z406.3→84.0(赖诺普利)和 m/z 349.1→206.1(内标依那普利拉)。结果:本文所建立测定赖诺普利的线性范围为1.064～851.2 ng·mL~(-1),最低定量限可达1.064 ng·mL~(-1)。测定的方法回收率为97.52%～103.2%;日内 RSD<7%,日间 RSD<5%。结论:本文所建立的方法灵敏度良好、准确度高,可用于赖诺普利的药代动力学研究及临床药物浓度监测。     

HPLC-MS／MS法测定动物源性食品中喹赛多的残留量

目的:建立了一种灵敏、快速的高效液相色谱-串联质谱法测定动物源性食品中喹赛多残留量。方法:样品用酸性乙腈-甲醇混合溶剂提取,经正己烷除脂,HLB 固相萃取小柱净化后,以水-乙腈-甲酸(70:30:0.1)为流动相,采用 YMC-Pack Pro C_(18)柱分离,用电喷雾正离子源进行离子化,多反应监测方式(MRM)对喹赛多的母离子(m/z 272)及其子离子(m/z188和143)进行检测。结果:本方法研究的喹赛多的线性范围为0～50 ng·mL~(-1),线性相关系数 r 大于0.9980;在10～40μg·kg~(-1)的3个添加水平范围内的平均回收率为69.10%～93.54%;相对标准偏差为2.49%～9.03%;方法检出限(LOD)为3μg·kg~(-1)。结论:该方法灵敏、准确、分析速度快,适用于喹赛多残留量检测和确证以及动力学的研究。     

HPLC-MS/MS法同时测定三七须根总皂苷中人参皂苷Rg_1和Re的含量

目的:建立以液-质联用法同时测定三七须根总皂苷中人参皂苷Rg1和Re含量的方法。方法:色谱柱为BDS HYPERSUL C18(150mm×2.1mm,5μm),流动相为乙腈-水(梯度洗脱),柱温为30℃;采用负离子多反应监测方法(MRM)测试。用于定量分析的对照品离子对分别为人参皂苷Rg1:m/z799.6→475.5;人参皂苷Re:m/z945.8→783.7;内标物紫杉醇m/z:852.5→525.3。结果:人参皂苷Rg1、人参皂苷Re标准曲线的线性范围分别为0.173～17.3μg·mL-1和0.156～39.1μg·mL-1,精密度和准确度等均符合样品分析的要求。结论:本方法准确、灵敏、特异性强,适用于三七须根总皂苷及其制剂中人参皂苷Rg1、Re含量的同时测定。     

HPLC／MS／MS法测定参麦注射液及人血浆中的人参皂苷Rg1

目的:为初步了解参麦注射液的药代动力学规律,建立高效液相色谱-串联质谱联用的分析方法,测定参麦注射液及人血浆中人参皂苷 Rg_1浓度。方法:色谱条件为 RESTEK PinnacleⅡ C_(18)柱(150mm×2.1mm,5μm),Phenomenex ODS Octade-cyl C_(18)保护柱(4mm×2.0mm);流动相:甲醇-水-乙腈(60∶20∶20);流速:250μL·min~(-1);柱温:20℃;质谱条件为气动辅助电喷雾离子源(ESI),检测方式为正离子多离子反应监测(MRM),用于定量分析的离子为 m/z 823.6→643.5;生物样品采用固相萃取处理。结果:参麦注射剂含量测定中人参皂苷 Rg_1的线性范围为5.25～525ng·mL~(-1),生物样品测定中人参皂苷 Rg_1的线性范围为1.02～1023ng·mL~(-1),精密度和准确度等均符合生物样品分析的要求。结论:该法专属、灵敏、准确,适用于参麦注射液和血浆中人参皂苷 Rg_1浓度的测定。     

脑得生片中10种成分的定量研究

目的建立HPLC法,对脑得生片中3′-羟基葛根素、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、染料木苷、三七皂苷Rb_1、人参皂苷Rg_1、大豆苷元和人参皂苷Rb_1 10种有效成分进行定量分析。方法色谱条件:色谱柱为Kromasil C_(18)柱(250 mm×4.6 mm,5μm);流动相为乙腈-2.0 g·L~(-1)磷酸水溶液,梯度洗脱;流速为0.8 mL·min~(-1);检测波长为250和203 nm;柱温为35℃。结果 10个成分的峰具有良好的分离度;3′-羟基葛根素、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、染料木苷、三七皂苷Rb_1、人参皂苷Rg_1、大豆苷元和人参皂苷Rb_1的线性范围分别为14.99～299.88,15.00～300.03,8.26～165.13,10.24～204.82,10.01～200.13,6.40～128.00,15.75～314.93,21.00～419.97,7.01～140.14和17.50～349.97μg·mL~(-1),相关系数分别为0.999 5,0.999 8,0.999 9,0.999 5,0.999 6,0.999 5,0.999 6,0.999 8,0.999 6和0.999 6;精密度、稳定性与重复性实验的RSD值均小于1.60%;10个对照品的加样回收率均在97.10%～98.48%之间,RSD值均在0.76%～1.56%之间;10批制剂中3′-羟基葛根素、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、染料木苷、三七皂苷Rb_1、人参皂苷Rg_1、大豆苷元和人参皂苷Rb_1的平均含量分别为7.43,13.33,8.29,10.64,9.35,1.04,2.21,8.44,1.73和2.97 mg·g~(-1),RSD值分别为1.23%,0.82%,1.11%,1.24%,1.07%,1.58%,1.28%,0.86%,1.45%和1.28%。结论脑得生片中10种主要有效成分的含量检测方法操作简便、准确、可靠、稳定且重复性好,为该制剂的质量控制提供了有效的评价方法。     

LC／ESI—MS测定小鼠血浆中异烟肼及其代谢物

目的:建立测定小鼠血浆中异烟肼及其代谢物乙酰肼、肼的液相色谱-电喷雾质谱联用(LC/ESI—MS)法。方法:血浆样品经甲醇沉淀蛋白后,用衍生化试剂3-甲氧基苯甲醛 MBA 进行柱前衍生化,衍生化产物以甲醇-5 mmol·L~(-1)醋酸铵(pH5.0)缓冲溶液(60:40)为流动相,采用 Agilent Zorbax XDB—C_8柱(5 μm,4.6 mm×150 mm)分离,通过 ESI~ -MS 以选择离子监测(SIM)模式进行检测,用于定量分析的离子分别为 m/z 256.1(异烟肼衍生物),m/z 193.2(乙酰肼衍生物),m/z 151.1(肼衍生物)。结果:异烟肼、乙酰肼、肼的线性范围分别为1～64,0.05～3.2,0.025～1.6μg·mL~(-1);定量下限分别为1,0.05,0.025μg·mL~(-1);方法的平均回收率分别为98.9%,98.0%,97.2%;平均 RSD 值分别为2.7%,2.4%,3.1%。结论:本法简便、准确,适用于小鼠血浆中异烟肼、乙酰肼和肼血药浓度的测定。     

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.     

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.     

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。     

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。