西他沙星代谢物的LC-MS法研究

采用 L C- MS技术分离和鉴定新氟喹诺酮类抗菌剂西他沙星体内代谢物。试验对口服西他沙星后大鼠、狗和人的尿样及胆汁分别进行分离和测定。结果在线分离和鉴定了西他沙星的 9个主要代谢物 (M1～M9) ,为西他沙星体内代谢研究及其它新氟喹诺酮类化合物的研制提供了良好的筛选结构群。     

HPLC-MS/MS法测定人尿液中西他沙星的浓度

目的：建立灵敏、快速、准确的人尿液中西他沙星浓度的HPLC-MS/MS检测方法,并将其用于测定健康受试者口服西他沙星颗粒剂后的尿药排泄。方法：尿液经甲醇处理后,取上清液进行HPLC-MS/MS法分析。10例受试者,每例空腹和餐后（随机交叉）单次口服西他沙星颗粒剂50 mg,经5天的清洗期后将两式受试者对调,两周期分别收集给药前（0 h）及给药后0~3、3~6、6~12、12~24、24~36、36~48、48~60 h各时间段的所有尿液并分析,计算西他沙星在尿液中的累计排泄量和累积排泄率。结果：西他沙星尿液的线性范围为0.250~200μg·mL-1,定量限为0.250μg· mL-1。健康受试者口服西他沙星颗粒剂后3 h即能检测到原型药物,36小时内经尿液排泄基本完全,累积排泄量约为35 mg,累积排泄率约为70%。结论：所建立的方法简便、可靠,可用于人口服西他沙星颗粒剂后药物尿液浓度的测定及其尿药排泄研究。     

HPLC-MS/MS在药物代谢和药动学筛选中的应用

目的本综述简要介绍先导化合物优化阶段,HPLC-MS／MS应用于药物代谢和药动学（DMPK）中筛选一系列新的化学实体。方法查阅有关文献,进行分析总结。结果HPLC-MS／MS在支持体内或体外试验以及定性分析,如代谢物鉴定等方面均有广泛应用。结论液质联用已成为DMPK筛选强有力且必不可少的分析手段。     

HPLC—MS／MS法与MEIA法在监测器官移植患者他克莫司全血浓度中的比较

目的：比较HPLC-MS／MS法与MEIA法在监测器官移植患者他克莫司全血浓度中的应用。方法：建立和确讧可行的HPLC-MS／MS法,并与ME—M法分别测定他克莫司相同样本的浓度,对测试结果进行统计分析,评价两种方法。结果：HPLC-MS肌S法测定他克莫司平均浓度为（4．86±0．46）ng／mL,MEIA法测定浓度平均为（5．52±0．43）ng／mL,两种方法测定结果相关系数r平均为0．8771,两种方法相关性较强。LC-MS／MS法测定浓度值／MEn法测定浓度值平均值为（90．34-5．3）％。结论：HPLC—MS／MS法测定浓度为他物在体内的准确浓度,更适用于日常治疗药物监测（TDM）工作。     

液相色谱-串联质谱与化学衍生化及在线氢／氘交换结合药物代谢物的特性策略

介绍了应用高效液相色谱．串联质谱（LC-MS／MS）与化学衍生化、在线氢／氘（H／D）交换技术联合研究粗提样品中药物代谢物结构的策略。讨论了子离子扫描、恒定中性丢失扫描、母离子扫描、多级的MS^n和精确质量测量的质谱扫描技术。尤其重要的是，只需微量的粗提样品就足以满足LC-MS／MS的分析要求，且可以免去从复杂生物样品中纯化低含量代谢产物的困难，从而减少工作强度。     

大鼠血浆中积雪甙浓度的HPLC/MS/MS测定

建立了HPLC／MS／MS测定大鼠血浆中的积雪甙。以柴胡皂甙D为内标，采用XTerra C18柱，流动相为O．1％乙酸和含O．1％乙酸的乙腈，梯度洗脱。质谱条件为电喷雾离子源，检测方式为多反应检测，定量分析离子为m／z981．5→493（积雪甙）和803→331（柴胡皂甙D），血浆样品采用固相萃取处理。检测限为0．5ng／ml。     

荷叶化学成分的HPLC-TOF/MS分析

目的通过高效液相-飞行时间质谱（HPLC-TOF／MS）联用技术定性分析荷叶中的主要化学成分。方法色谱分离采用AgilentZorbaxSB-Aq（250mmX4．6mm,5μm）,流动相组成分别为乙腈和1．0％醋酸,梯度洗脱,流速为1．0ml／min,质谱定性采用飞行时间质谱,正离子模式扫描。结果在优化的液质联用条件下,结合数据库匹配技术和碎片离子分析,鉴定出荷叶中主要的生物碱类和黄酮类成分。结论建立了一种简单、可靠的HPLC-TOF／MS方法对荷叶中主要化学成分进行了鉴定。     

LC-MS／MS法测定人血浆中的帕珠沙星

目的：建立一种快速灵敏检测帕珠沙星血药物浓度的液相色谱－串联质谱方法。方法色谱柱为 Agilent Eclipse XDB－C18（4．6mm ×150mm,5μm）;流动相为乙腈：水（0．005mol· L－1甲酸铵,0．1％甲酸）＝（30∶70）;流速：0．5mL· min －1;质谱条件为电喷雾电离源（ESI）;采用选择反应监测（SRM）对帕珠沙星（m／z 319．0→301．1）和环丙沙星（m／z 332．2→314．2）进行测定。样本处理采用乙腈沉淀蛋白法,吸取上清液1μL进样。结果帕珠沙星的线性范围为0．02～20μg· mL －1,提取回收率为82％～92％,日内和日间精密度均小于15％。结论本方法简便、灵敏,该方法经考察符合血浆样品的测定要求,可以应用于临床药物浓度的测定和药代动力学的研究。     

用细胞膜色谱法和HPLC-TOF/MS研究附子中的有效成分

目的通过细胞膜色谱技术和高效液相-飞行时间质谱（HPLC-TOF／MS）技术相结合,初步筛选和鉴定附子中的有效成分。方法初步筛选采用自制的大鼠心肌细胞膜色谱柱,流动相为10mmol／L磷酸盐缓冲溶液,流速为0．2ml／min。色谱分离采用SHISEIDO CAPCELL PAKC18柱,流动相为0．1％甲酸水溶液和乙腈,梯度洗脱,流速为1．0ml／min。质谱鉴定采用TOF／MS,正离子模式扫描。结果确定了附子中8个可能的有效成分。结论通过细胞膜色谱技术和HPLC-TOF／MS技术相结合,为从天然产物中快速寻找新的活性成分提供了途径。     

LC-MS／MS在中毒检测中的应用及病例分析

目的：建立血液中常见毒药物的快速、高效LC—MS／MS筛查定性方法。方法：采用固相萃取的前处理方法,通过IDA扫描模式,运用多反应监测作筛查,再依赖3个不同能量的增强型二级扫描,最终在数据库中筛选中毒药物。结果：运用LC—MS／MS快速筛查已成功为43例患者进行毒物检测分析,为临床抢救赢得了宝贵的时间、提供了有力的依据。结论：本方法简单、快速、准确,可用于中毒患者的毒物药物定性分析。     

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).     

LC/MS/MS analysis of vesnarinone and its principal metabolites in plasma and urine

An LC/MS/MS assay was developed and successfully used to quantitate vesnarinone and its principal metabolites (OPC-8230, OPC-18136, and OPC-18137) in human plasma and urine. Samples were pre-treated with liquid–solid extraction followed by simultaneous monitoring of primary and daughter ions which were used for the identification and quantitation of the analytes on LC/MS/MS. This assay offers advantages of specificity, speed and greater sensitivity over the previously developed HPLC-UV assay. The lower limit of quantitation is 500 ng ml⁻¹ for vesnarinone and 20 ng ml⁻¹ for OPC-8230, OPC-18137, and OPC-18136 in plasma. Methodology is similar for the estimation of these analytes in urine with the lower limit of quantitation being 500 ng ml⁻¹ for vesnarinone and 100 ng ml⁻¹ for each metabolite. Ascorbic acid was added to stabilize the analytes from degradation. This LC/MS/MS method was developed to overcome many practical problems associated with the HPLC method. The LC/MS/MS method offers the flexibility of analyzing additional metabolites and changing the linearity range to accommodate the differences in linear range (200–10 000 ng ml⁻¹ for vesnarinone and 20–1000 for metabolites) for the analytes.     

LC‐MS/MS detection of unaltered glucuronoconjugated metabolites of metandienone

The aim of this study was to evaluate the direct detection of glucuronoconjugated metabolites of metandienone (MTD) and their detection times. Metabolites resistant to enzymatic hydrolysis were also evaluated. Based on the common mass spectrometric behaviour of steroid glucuronides, three liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) strategies were applied for the detection of unpredicted and predicted metabolites: precursor ion scan (PI), neutral loss scan (NL), and theoretical selected reaction monitoring (SRM) methods. Samples from four excretion studies of MTD were analyzed for both the detection of metabolites and the establishment of their detection times. Using PI and NL methods, seven metabolites were observed in post‐administration samples. SRM methods allowed for the detection of 13 glucuronide metabolites. The detection times, measured by analysis with an SRM method, were between 1 and 22 days. The metabolite detected for the longest time was 18‐nor‐17β‐hydroxymethyl‐17α‐methyl‐5β‐androsta‐1,4,13‐triene‐3‐one‐17‐glucuronide. One metabolite was resistant to hydrolysis with β ‐glucuronidase; however it was only detected in urine up to four days after administration. The three glucuronide metabolites with the highest retrospectivity were identified by chemical synthesis or mass spectrometric data, and although they were previously reported, this is the first time that analytical data of the intact phase II metabolites are presented for some of them. The LC‐MS/MS strategies applied have demonstrated to be useful for detecting glucuronoconjugated metabolites of MTD, including glucuronides resistant to enzymatic hydrolysis which cannot be detected by conventional approaches. Copyright © 2016 John Wiley & Sons, Ltd.     

UHPLC‐MS/MS and UHPLC‐HRMS identification of zolpidem and zopiclone main urinary metabolites and method development for their toxicological determination

Zolpidem and zopiclone (Z‐compounds) are non‐benzodiazepine hypnotics of new generation that can be used in drug‐facilitated sexual assault (DFSA). Their determination in biological fluids, mainly urine, is of primary importance; nevertheless, although they are excreted almost entirely as metabolites, available methods deal mainly with the determination of the unmetabolized drug. This paper describes a method for the determination in urine of Z‐compounds and their metabolites by ultra‐high‐pressure liquid chromatography/tandem mass spectrometry (UHPLC‐MS/MS) and UHPLC coupled with high resolution/high accuracy Orbitrap® mass spectrometry (UHPLC‐HRMS). The metabolic profile was studied on real samples collected from subjects in therapy with zolpidem or zopiclone; the main urinary metabolites were identified and their MS behaviour studied by MS/MS and HRMS. Two carboxy‐ and three hydroxy‐ metabolites, that could be also detected by gas chromatography/mass spectrometry (GC‐MS) as trimethylsylyl derivatives, have been identified for zolpidem. Also, at least one dihydroxilated metabolite was detected. As for zopiclone, the two main metabolites detected were N‐demethyl and N‐oxide zopiclone. For both substances, the unmetabolized compounds were excreted in low amounts in urine. In consideration of these data, a UHPLC‐MS/MS method for the determination of Z‐compounds and their main metabolites after isotopic dilution with deuterated analogues of zolpidem and zopiclone and direct injection of urine samples was set up. The proposed UHPLC‐MS/MS method appears to be practically applicable for the analysis of urine samples in analytical and forensic toxicology cases, as well as in cases of suspected DFSA. Copyright © 2013 John Wiley & Sons, Ltd.     

液相色谱-电喷雾离子阱质谱法研究人尿中1,5-二咖啡酰奎宁酸的代谢产物

目的 研究健康受试者口服1,5-二咖啡酰奎宁酸后尿液中的代谢产物.方法 健康受试者每人口服1,5-二咖啡酰奎宁酸600 mg,收集0-24 h的尿样,经C_(18)小柱固相萃取纯化后,用液相色谱-电喷雾离子阱质谱联用技术对人尿中的代谢产物进行分析鉴定.结果 在人尿中发现了1,5-二咖啡酰奎宁酸的甲基化、葡萄糖醛酸化及甲基-葡萄糖醛酸化代谢产物共28个.其中,有2个代谢产物结构经标准品对照得到确证.结论 甲基化、葡萄糖醛酸化和异构化反应是1,5-二咖啡酰奎宁酸在人体内的3种重要代谢途径.     

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.     

高效液相色谱-电喷雾离子阱串联质谱分析水苏碱及其大鼠体内代谢物

目的鉴定水苏碱在大鼠体内的代谢物。方法应用高效液相色谱-电喷雾离子阱串联质谱(HPLC-ESI/MSⁿ)技术研究水苏碱的一级质谱电离规律、二级质谱裂解规律及其色谱保留，以此作为水苏碱大鼠体内代谢物分析鉴定的依据；再将健康大鼠空腹灌胃25 mg·kg^-1水苏碱，收集0～24 h的尿样，经C₁₈小柱固相萃取分离纯化后，应用HPLC-ESI/MS分析尿样中水苏碱代谢物。结果在大鼠尿样中发现了母药及其N-去甲基、氧化脱氢、环氧化等6种I相代谢产物及两种环氧化物的甘氨酸轭合II相代谢产物。结论HPLC-ESI/MS法灵敏度高，快速，定性能力强，适合于水苏碱大鼠体内代谢物的分析。     

液相色谱-串联质谱法测定人血浆中兰索拉唑及其代谢物浓度

目的建立液相色谱-串联质谱同时测定人血浆中兰索拉唑及其代谢物兰索拉唑砜、5-羟基兰索拉唑浓度的方法。方法用Agilent SB-C18色谱柱,流动相为0.002 mol.L-1乙酸铵(用甲酸调pH为4)-乙腈(60∶40,v/v),流速为0.3 mL.min-1。用正离子电离,多离子反应监测进行定量分析。结果兰索拉唑、兰索拉唑砜、5-羟基兰索拉唑线性范围分别为5～3044,1.5～550,2～549.5 ng.mL-1,三者日内、日间精密度均小于10%,三者的提取回收率为81.55%～98.74%(RSD<10%)。结论本方法灵敏、准确、快速,可用于人血浆中兰索拉唑及其代谢物浓度的测定和药代动力学研究。     

Pharmacokinetics and metabolism of lithospermic acid by LC/MS/MS in rats

The pharmacokinetics and metabolism of lithospermic acid (LA), a component isolated from Salvia miltiorrhiza, and its two O-methylated metabolites (3′-monomethyl- and 3′,3″-dimethyl-lithospermic acid), were analyzed by a rapid and specific isocratic liquid chromatography-tandem mass spectrometry (LC/MS/MS) method. Rat serum samples collected after intravenous and oral administration were analyzed for obtaining pharmacokinetic data of LA. Two O-methylated metabolites, namely one 3′-monomethyl- and one 3′,3″-dimethyl-lithospermic acid were detected in rat serum and bile samples after intravenous and oral administration of LA, respectively. An oral bioavailability of 1.15% was found, with the AUC_0–t values of 301.89 and 3.46 mg h/L for intravenous and oral administration, respectively. The total recovery from bile was 75.36% (0.46% for LA, 17.23% for M1, and 57.67% for M2) after intravenous administration, and 4.26% (0.00% for LA, 0.10% for M1, and 4.16% for M2) after oral administration. These results indicate that methylation is the main metabolic pathway of LA, and that LA is excreted into rat bile and finally into feces.     

A UPLC–MS/MS application for profiling of intermediary energy metabolites in microdialysis samples—A method for high-throughput

Research within the field of metabolite profiling has already illuminated our understanding of a variety of physiological and pathological processes. Microdialysis has added further refinement to previous models and has allowed the testing of new hypotheses. In the present study, a new ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS/MS) method for the simultaneous detection and quantification of intermediary energy metabolites in microdialysates was developed. The targeted metabolites were mainly from the citric acid cycle in combination with pyruvic acid, lactic acid, and the ATP (adenosine triphosphate) hydrolysis product adenosine along with metabolites of adenosine. This method was successfully applied to analyze the microdialysates obtained from an experimental animal study giving insight into the hitherto unknown concentration of many interstitial energy metabolites, such as succinic acid and malic acid. With a total cycle time of 3 min, injection to injection, this method permits analysis of a much larger number of samples in comparison with conventional high performance liquid chromatography/tandem mass spectrometry HPLC–MS/MS strategies. With this novel combination where microdialysis and high sensitivity UPLC–MS/MS technique is combined within cardiologic research, new insights into the intermediary energy metabolism during ischemia–reperfusion is now feasible.