避孕药及其生物样品定量测定方法的进展

介绍了避孕药的发展状况及其在生物样品中的定量测定方法.现有放射免疫分析、GC-MS法和LC-MS/MS法的灵敏度均较高,其中LC-MS/MS法成为生物样品测定的主要方法.     

液-质联用中大气压电离接口技术及其在药物分析中的应用

目的：介绍液相色谱-质谱联用技术（LC-MS）在药物分析中的应用。方法：通过介绍液相色谱-质谱技术原理，引入大气压电离接口技术，并综述了在药物分析中的应用。结果：液相色谱-质谱技术对药物的杂质检查与降解产物、药动学、药物代谢产物的分析和鉴定、生物大分子以及药物开发得到了广泛应用。结论：大气压电离接口技术，扩大了液相色谱-质谱联用技术的应用范围，促进了药物分析学科的发展。     

生物分析中溶血对检测的影响及其对策

液相色谱-串联质谱(LC-MS/MS)技术灵敏度高,选择性强,是目前体内药物分析的首选方法 ,但LC-MS/MS生物分析中仍面临着溶血样品测定失败的难题。溶血导致的样品测定数据不准确,将直接影响药物的药动学研究,如何进行溶血样品的测定已成为各国监管机构和全球生物分析行业探讨的重点。本文从溶血样品产生的原因进行分析,将溶血对检测的影响和作用方式以及溶血效应的考察方法进行了概述,最后对溶血样品的测定方法和策略进行探讨。     

LC-MS/MS法测定人血浆中3种酪氨酸激酶抑制剂的浓度及应用

目的 建立一种液相色谱-串联质谱(LC-MS/MS)法,可同时检测人血浆中3种表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)的血浆药物浓度。方法 采用甲醇作为沉淀剂,对血浆样本进行前处理,采用Kinetex XB-C₁₈(50.0 mm×3.0 mm,2.6μm)进行色谱分离,梯度洗脱采用含0.1%甲酸的水溶液与乙腈溶液,柱温40℃,流速0.3 m L·min^-1,进样量1μL。采用电喷雾正电离模式多反应监测模式,进行血浆药物浓度检测和定量分析。结果 色谱分离和质谱检测的单针分析时长为5.5 min,奥希替尼、阿美替尼和伏美替尼的血浆药物浓度分别在2.5～500.0、2.5～500.0和0.5～100.0 ng·m L^-1内显示出良好的线性关系,相关系数均>0.999 0。结论 该研究建立的LC-MS/MS法灵敏度和准确度均符合生物样品定量分析方法验证指导原则,同时符合生物样品定量分析指南,具有样本进样体积小、分析样本时间少等技术优势,可用于患者血浆样本的分析和监测。     

寡核苷酸药物及生物标志物的生物分析研究进展

寡核苷酸已成为多种疾病诊断和治疗的候选药物。寡核苷酸药物及其代谢物、生物标志物的定性定量分析是药物开发和评估所必需的,良好的分析方法有利于控制药品质量及准确定量药品浓度。本文主要探讨了逆转录-实时荧光定量PCR (RT-qPCR)、液相色谱-荧光(LC-FL)、液相色谱-质谱联用（LC-MS/MS）、液相色谱-高分辨质谱联用（LC-HRMS）在寡核苷酸药物和生物标志物的应用及各自的优缺点,旨在概述目前现有寡核苷酸药物和生物标志物的生物分析方法,以期为从事寡核苷酸诊断和治疗药物的研发、分析人员及企业提供参考,以期提高此类新药或仿制药一致性评价分析的水平。     

HPLC和LC-MS/MS鉴定锁阳固精丸中非法添加的西地那非衍生物

目的 鉴定中成药中非法掺入的西地那非衍生物。方法 采用高效液相色谱-二极管阵列检测器法(HPLC-DAD)和液相色谱-电喷雾电离-质谱联用法(LC-ESI-MS/MS),定性分析锁阳固精丸中非法掺入的西地那非衍生物。结果 通过比较被测样品成分和西地那非对照品的紫外吸收光谱图,并对两者的质谱碎片数据进行解析和研究,确认西地那非的一种衍生物羟甲基西地那非(hydroxyhomosildenafil)。结论 利用液相色谱-质谱联用法(LC-MS/MS),结合质谱分子结构解析技术,具有专属性强的特点,可以准确分析中成药中非法添加的西地那非衍生物。     

我院乳腺手术患者围手术期抗菌药物预防应用的调查分析

目的鉴定中成药中非法掺入的西地那非衍生物。方法采用高效液相色谱-二极管阵列检测器法(HPLC-DAD)和液相色谱-电喷雾电离-质谱联用法(LC-ESI-MS/MS),定性分析锁阳固精丸中非法掺入的西地那非衍生物。结果通过比较被测样品成分和西地那非对照品的紫外吸收光谱图,并对两者的质谱碎片数据进行解析和研究,确认西地那非的一种衍生物羟甲基西地那非(hydroxyhomosildenafil)。结论利用液相色谱-质谱联用法(LC-MS/MS),结合质谱分子结构解析技术,具有专属性强的特点,可以准确分析中成药中非法添加的西地那非衍生物。     

LC-MS/MS法和MEIA法测定人血浆中地高辛浓度的差异比较

目的:比较液相色谱-串联质谱(LC-MS/MS)法与微粒子酶联免疫(MEIA)法测定人血浆中地高辛浓度的差异。方法:分别采用LC-MS/MS法与MEIA法对质控样品及50例患者血浆样品中的地高辛浓度进行检测,采用配对t检验比较分析两种方法的检测结果。结果:质控样品检测结果显示,两种方法检测结果间的差异无统计学意义(P>0.05);50例患者血浆样品检测结果显示,MEIA法检测结果明显高于LC-MS/MS法,两者差异有统计学意义(P<0.01)。结论:MEIA法检测结果与临床实际情况相比存在一定的差异,LC-MS/MS法更适于临床地高辛血药浓度的检测。     

妇科止带片中金胺O的检测方法研究

目的建立高效液相色谱(HPLC-PDA)法和液相色谱-质谱联用(LC-MS/MS)法对妇科止带片中可能掺入的化工染料金胺O进行测定。方法采用HPLC-PDA法对妇科止带片样品进行筛查,采用LC-MS/MS法对筛查结果进行验证。结果市售的25批样品中,有5批检出了金胺O。结论建立的检查方法准确、可靠、专属性强,可有效检测妇科止带片中金胺O的染色情况。     

微探针电喷雾串联质谱与液相色谱-电喷雾串联质谱的基质效应对比研究

电喷雾电离(electrospray ionization, ESI)易受基质干扰,影响其定量的准确度、精密度和稳定性。探针电喷雾电离(probe electrospray ionization, PESI)是原位电离的代表之一,无色谱分离,无复杂样品前处理,具有简便、快速、高通量等诸多优点。微探针电喷雾串联质谱(micro pen electrospray ionization tandem mass spectrometry,μPen-ESI-MS/MS)是基于现有的PESI开发的新技术。本文旨在评价μPen-ESI-MS/MS方法用于血浆中药物定量分析的基质效应,并与液相色谱-电喷雾串联质谱(liquid chromatography coupled with electrospray ionization tandem mass spectrometry, LC-ESI-MS/MS)方法的基质效应进行比较。分别建立5种药物血浆样品的μPen-ESI-MS/MS和LC-ESI-MS/MS方法,计算两种方法的基质因子及内标归一化的基质因子。结果表明,他克莫司、氟桂利嗪和地氯雷他定的...     

Determination of okadaic acid, dinophysistoxin-1 and related esters in Greek mussels using HPLC with fluorometric detection, LC-MS/MS and mouse bioassay

An approach involving both chemical and biological methods was undertaken for the detection and quantification of the marine toxins okadaic acid (OA), dinophysistoxin-1 (DTX-1) and their respective esters in mussels from different sampling sites in Greece during the period 2006-2007. Samples were analyzed by means of a) high performance liquid chromatography with fluorometric detection (HPLC-FLD), using 9-athryldiazomethane (ADAM), as a pre-column derivatization reagent, b) liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and c) the mouse bioassay. Free OA and DTX-1 were determined by both HPLC-FLD and LC-MS/MS, while their respective esters were determined only by LC-MS/MS after alkaline hydrolysis of the samples. The detection limit (L.O.D.) and quantification limit (L.O.Q.) of the HPLC-FLD method were 0.015 μg/g HP and 0.050 μg/g HP, respectively, for OA. The detection limit (L.O.D.) and quantification limit (L.O.Q.) of the LC-MS/MS method were 0.045 μg/g HP and 0.135 μg/g HP, respectively, for OA. Comparison of results between the two analytical methods showed excellent agreement (100%), while both HPLC-FLD and LC-MS/MS methods showed an agreement of 97.1% compared to the mouse bioassay.     

LC-MS/MS技术及其在天然产物分析中的应用

目的:研究LC-MS/MS技术及其在天然产物分析中的应用;方法:通过分析Lc-MS接口的基本原理详细研究LC-MS/MS技术,并分别介绍其在天然产物分析中微量天然物质分析、天然产物异构体的分离和鉴定、天然产物筛选和中药指纹图谱四个方面的应用.结果和结论:LC-MS/MS在技术及应用方面取得了很大进展,相信在天然产物分析中必将发挥越来越重要的作用.     

LC-MS/MS技术在蛋白质药物定量分析中的应用

近年来，蛋白质药物已经成为全球制药业的研发热点。建立灵敏、准确、高通量的蛋白质药物定量分析方法，是进行蛋白质药物研究的关键前提，也是蛋白质药物研究的难点。随着液相色谱串联质谱技术（LC-MS/MS）的发展，其在蛋白质药物定量分析中起着越来越重要的作用。本文首先综述了蛋白质药物的分类，然后比较了LC-MS/MS技术与传统配体结合测定方法（LBA）在蛋白质药物分析中的优缺点，以及分析了LC-MS/MS技术在蛋白质药物定量分析中的常见问题、解决方法、应用方向等。最后预测，短时间内LC-MS/MS 不会完全取代传统的LBA方法，随着LC-MS/MS技术和生物化学技术的充分结合，蛋白质药物分析将会得到飞速的发展。     

LC-MS/MS法测定大鼠血浆中吴茱萸碱的浓度及其药代动力学研究

目的：建立用于测定吴茱萸碱血药浓度的液相色谱-串联质谱联用分析方法,并研究吴茱萸碱在大鼠体内的药代动力学。方法：6只大鼠灌胃给药吴茱萸碱100mg/kg,眼底取血,LC-MS/MS法测定血药浓度,并用DAS药代动力学程序拟合计算药代动力学参数。结果：吴茱萸碱浓度在0.2～50ng/mL内,线性关系良好（r^2=0.9997）。提取回收率96.12%～99.46%,日内、日间RSD分别为4.61%～13.51%和5.65%～11.49%。主要药代动力学参数为：Cmax=（5.3±1.5）ng/mL;tmax=（22±8）min;t1/2=（451±176）min。结论：建立的LC-MS/MS方法专属性强,灵敏度高,可用于吴茱萸碱的体内定量分析。     

Sensitive LC-MS/MS methods for the quantification of RGH-188 and its active metabolites, desmethyl- and didesmethyl-RGH-188 in human plasma and urine

Selective and sensitive LC-MS/MS methods have been developed and validated for simultaneous determination of RGH-188, a novel atypical antipsychotic, and its two active metabolites, desmethyl- and didesmethyl-RGH-188 in human plasma and urine. Deuterated analytes, [2H6]-RGH-188, [2H3]-desmethyl-RGH-188 and [2H8]-didesmethyl-RGH-188 were used as internal standards (IS). The compounds were isolated from the alkalized biological matrix using liquid-liquid extraction (LLE) and the extracts were analysed by reversed-phase HPLC with MS/MS detection. The chromatographic run time was 5.0min per injection. The PE Sciex API 365 mass spectrometer was equipped with a TurboIonSpray interface and operated in positive-ion, multiple reaction monitoring (MRM) mode. The mass transitions monitored were m/z 427.3-->382.2, 413.2-->382.2, 399.2-->382.2, 433.3-->382.2, 416.2-->382.2 and 407.3-->390.2 for RGH-188, desmethyl-RGH-188, didesmethyl-RGH-188, [2H6]-RGH-188, [2H3]-desmethyl-RGH-188 and [2H8]-didesmethyl-RGH-188, respectively. The lower limit of quantification (LLOQ) was 0.05 and 0.1ng/ml for RGH-188 and its metabolites, respectively, using 1ml of plasma. LLOQ in 1ml of urine was 0.1ng/ml for all three analytes. The methods were validated for selectivity, linearity, accuracy and precision. The lower limit of quantification, dilution integrity, matrix effect, stability of the analytes in the biological matrix during short- and long-term storage and after three freeze-thaw cycles were also tested. The assays were simple, specific and robust enough to support clinical development of RGH-188.     

Development and validation of an LC‐MS/MS method after chiral derivatization for the simultaneous stereoselective determination of methylenedioxy‐methamphetamine (MDMA) and its phase I and II metabolites in human blood plasma

3,4‐Methylenedioxymethamphetamine (MDMA, ecstasy) is a racemic drug of abuse and its two enantiomers are known to differ in their dose‐response curves. The S‐enantiomer was shown to be eliminated at a higher rate than the R‐enantiomer. The most likely explanation for this is a stereoselective metabolism also claimed in in vitro studies. Urinary excretion studies showed that the main metabolites in humans are 4‐hydroxy 3‐methoxymethamphetamine (HMMA) 4‐sulfate, HMMA 4‐glucuronide and 3,4‐dihydroxymethamphetamine (DHMA) 3‐sulfate. For stereoselective pharmacokinetic analysis of phase I and phase II metabolites in human blood plasma useful analytical methods are needed. Therefore the aim of the presented study was the development and validation of a stereoselective liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method for the simultaneous quantification of MDMA, 3,4‐methylenedioxyamphetamine, DHMA, DHMA 3‐sulfate, HMMA, HMMA 4‐glucuronide, HMMA 4‐sulfate, and 4‐hydroxy 3‐methoxyamphetamine in blood plasma for evaluation of the stereoselective pharmacokinetics in humans. Blood plasma samples were prepared by simple protein precipitation and afterwards all analytes were derivatized using N‐(2,4‐dinitro‐5‐fluorophenyl) L‐valinamide resulting in the formation of diastereomers which were easily separable on standard reverse phase stationary phases. This simple and fast method was validated according to international guidelines including specificity, recovery, matrix effects, accuracy and precision, stabilities, and limits of quantification. The method proved to be selective, sensitive, accurate and precise for all tested analytes except for DHMA. Copyright © 2014 John Wiley & Sons, Ltd.     

Determination of fexofenadine in human plasma by LC-MS/MS and its application in pharmacokinetic study

This study presented a simple, rapid, and sensitive liquid chromatography analytical method employing tandem mass spectrometry (LC-MS/MS) to determine fexofenadine in human plasma. After the de-proteination procedure with acetonitrile, chromatographic separation of fexofenadine was performed using a reversed-phase Eclipse XDB-C₈ column with a mobile phase consisted of 1 mmol/L ammonium acetate buffer solution containing 0.2% formic acid-methanol (45:55, v/v). Fexofenadine was quantified using tandem mass detection in the electrospray ionization (ESI) positive ion mode. The flow rate of the mobile phase was 1 mL/min, and the retention times of fexofenadine and the internal standard (IS, losartan) were 1.76 min and 2.65 min, respectively. The calibration curve was linear over the plasma concentration range of 1-1000 ng/mL. The relative standard deviations of intra- and inter-batches were less than 10.4% and 15.4%, respectively. The LC-MS/MS method reported in this study showed higher sensitivity for the quantification of fexofenadine in human plasma than that shown by previously described analytical methods. Lastly, the method was successfully applied to the pharmacokinetic of fexofenadine in healthy Taiwan volunteers.     

Development and validation of an analytical method based on high performance thin layer chromatography for the simultaneous determination of lamotrigine, zonisamide and levetiracetam in human plasma

Methods based on HPLC technology are the most frequently adopted for monitoring blood levels of novel antiepileptics. Here a rapid method based on HPTLC was developed for quantitative determination of lamotrigine (LTG), zonisamide (ZNS) and levetiracetam (LVT) in human plasma and compared with HPLC and LC-MS/MS methods. Chromatographic separation was achieved on silical gel 60F₂₅₄ plates using ethylacetate:methanol:ammonia (91:10:15 v/v/v) as mobile phase. Quantitative analysis was carried out by densitometry at a wavelength of 312, 240 and 210 nm for LTG, ZNS and LVT, respectively. Calibration curves were linear over range of 0-200 ng for LTG and ZNS and 0-400 ng for and LVT. The limit of quantification of LTG, ZNS and LTV was found to be 3.69, 3.7 and 6.85 μg/ml, respectively. Intra and inter-assay precision provided relative standard deviations lower than 10% for all three analytes. Correlation and Bland-Altman plot showed general agreement between HPTLC and LC-MS/MS quantification, with a mean bias of −0.25, −0.46 and 0.5 μg/ml for LTG ZNS and LVT, respectively. Likewise, comparison between HPLC-UV and LC-MS/MS showed good agreement for all the three compounds analyzed. In conclusion, the proposed HPTLC method is simple, rapid, precise and accurate. It therefore is appropriate for the routine quantification of therapeutic levels of LTG, ZNS and LVT in human plasma.     

Supercritical fluid chromatography-tandem mass spectrometry for fast bioanalysis of R/S-warfarin in human plasma

Chiral separation for the analysis of enantiomers in biological fluids by HPLC often takes relatively long chromatography time compared to achiral analysis. The advantage of fast mass transfer in packed-column supercritical fluid chromatography (pSFC) and the high-flow compatibility of APCI-MS/MS were applied to develop a fast bioanalytical method for R/S-warfarin in human plasma. Presented here are the main challenges encountered during method development of a semi-automated liquid extraction SFC-MS/MS method. The selection of internal standard, robustness of the SFC equipment, and carryover issues are discussed. The method has high-throughput: the chromatography time is at least two-fold faster than the our fastest previous method; and the liquid/liquid extraction time of 96 samples is less than 20 min using a Tecan Genesis^® RSP 100 pipetting station and a Tomtec Quadra-96^® workstation. The standard curve range was 13.6–2500 ng/ml. Precision of QC concentrations from four validation runs was 7.0% for R-warfarin and 6.0% C.V. for S-warfarin; and the bias was 3.7 and 3.2% R.E., respectively. The method is sensitive, accurate, selective and robust, and was applied to a drug-interaction clinical study with rapid turnaround of sample analysis.     

Recent applications of liquid chromatography-mass spectrometry in natural products bioanalysis

Natural flavonoids, alkaloids, saponins and sesquiterpenoids have been extensively investigated because of their biological and physiological significances, as well as their promising clinical uses. It is necessary to monitor them or their metabolites in biological fluids for both pre-clinical studies and routine clinical uses. The successful hyphenation of LC and MS, which was thought as "the bird wants to marry with fish", has been conducted widely in biological samples analysis. This present paper reviewed the feasibility of LC-MS techniques in the identification and quantification of natural products (flavonoids, alkaloids, saponins and sesquiterpenoids) in biological fluids, dealing with sample preparation, LC techniques, suitability of different MS techniques. Perspective of LC-MS was also discussed to show the potential of this technology. The citations cover the period 2002-2006. We conclude that LC-MS is an extremely powerful tool for the analysis of natural products in biological samples.