液相色谱-质谱联用技术在药物代谢产物鉴定中的应用

药物代谢研究贯穿药物开发的整个过程。生物基质中药物代谢产物的快速、准确鉴定有助于理解药物或候选化合物的生物转化途径,确定代谢软位点,从而帮助优化先导物结构,筛选出具有更高代谢稳定性的药物。利用亲核性捕获试剂开展体外代谢实验,检测反应性代谢产物的生成,相关结果可以帮助规避候选化合物进入临床后可能产生的毒性风险。由于液相色谱-质谱联用技术（LC/MS）具有分析通量高、检测灵敏度高、选择性好、能提供丰富的结构相关信息等优点,其在药物代谢研究中的作用越来越重要。本工作综述了近年来作者所在实验室采用LC/MS法开展的药物代谢产物鉴定研究,提出了基于LC/MS技术的药物代谢产物鉴定研究的基本流程和药物在人体内的主要代谢产物,可为临床药动学研究和药物相互作用研究提供数据基础。     

划时代的高分辨质谱——安捷伦6550 iFunnel四极杆-飞行时间液相色谱/质谱联用仪

本文主要介绍了全新的安捷伦6550 iFunnel四极杆-飞行时间液相色谱/质谱联用仪。安捷伦6550 iFunnel Q-TOF引入了划时代的安捷伦iFunnel技术,以无可比拟的速度和灵敏度,实现一台仪器上同时完成最具挑战性的定性和定量分析。6550 iFunnel Q-TOF集突破性的iFunnel技术、离子束压缩冷却技术（IBCS）技术、喷射流离子聚焦技术Jet Stream4、GHz高速检测器和模数转换采集模式（ADC）和双增益放大器等最强创新技术于一身,打造出同时具有高分离度、超高质量精度和飞克级灵敏度的高分辨质谱仪。     

Liquid chromatography–tandem mass spectrometry applications in endocrinology

Liquid chromatography–tandem mass spectrometry (LC–MS/MS) has been recognized as a primary methodology for the accurate analysis of endogenous steroid hormones in biological samples. This review focuses on the use of LC–MS/MS in clinical laboratories to assist with the diagnosis of diverse groups of endocrine and metabolic diseases. Described analytical methods use on‐line and off‐line sample preparation and analytical derivatization to enhance analytical sensitivity, specificity, and clinical utility. Advantages of LC–MS/MS as an analytical technique include high specificity, possibility to simultaneously measure multiple analytes, and the ability to assess the specificity of the analysis in every sample. All described analytical methods were extensively validated, utilized in routine diagnostic practice, and were applied in a number of clinical and epidemiological studies, including a study of the steroidogenesis in ovarian follicles. © 2009 Wiley Periodicals, Inc. Mass Spec Rev 29:480‐502, 2010     

nanoUPLCQ-TOFMS/MS在大鼠肝移植蛋白质组学研究中潜在生物标记物的应用

建立应用纳升级超高效液相色谱 质谱（nanoUPLC Q-TOF MS/MS）联用技术研究大鼠同位肝移植术后不同时间差异表达蛋白质的蛋白质组学方法。建立10组大鼠肝移植动物模型后,随机分成术后3天组（n=5）和术后7天组（n=5）。分别于术后3天和7天处死5只受体,取肝组织,提取总蛋白,Trypsin酶解后,使用Waters公司独家专利MSe^TM无标记定量技术,利用nanoUPLC-MS/MS技术对酶解后的肽段进行分析,数据库检索鉴定蛋白质,采用生物信息学工具对所鉴定的蛋白质进行功能分类。7天组与3天组相比,两样品中同时存在的大部分蛋白上调,其中10个蛋白点表达水平明显上调,比值变化在8.5倍以上。结合数据库搜索得到鉴定,这些明显上调的蛋白的分子功能主要与细胞骨架、离子结合、氧化应激反应、能量代谢等相关。     

ESQUIRE-LC离子阱LC/MS~n

ESQUIRE-LC离子阱LC/MS~n是将惠普(HP)与布鲁克(Bruker)的尖端技术相结合的高效液相色谱/质谱(LC/MS~n)系统。惠普公司成功的1100系列HPLC和先进的电喷雾电离/大气压化学电离(ESI/APCI)离子源与布鲁克公司最新的多极离子阱MS和MS~n分析器完美结合在一起。ESQUIRE-LC是一种先进的自动化程度最高的液质联用仪。本文是对该仪器的性能特点、技术指标及应用的简要介绍。     

Environmental and food applications of LC-tandem mass spectrometry in pesticide-residue analysis: an overview

An overview is given on pesticide-residue determination in environmental and food samples by liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS). Pesticides comprise a large number of substances that belong to many completely different chemical groups, the only common characteristic is that they are effective against pests. They still constitute a challenge in MS because there is no collective pathway for fragmentation. A brief introduction to the theory of tandem MS permits a discussion of which parameters influence the ionization efficiency when the ions are subjected to different actions. Emphasis is placed on the different tandem MS instruments: triple and ion-trap quadrupoles, and hybrid quadrupole time-of-flight (Q-TOF), including advantages and drawbacks, typical detection limits, and ion signals at low concentrations. The instrumental setup, as well as LC and mass spectrometric experimental conditions, must be carefully selected to increase the performance of the analytical system. The capacity of each instrument to provide useful data for the identification of pesticides, and the possibility to obtain structural information for the identification of target and non-target compounds, are discussed. Finally, sample preparation techniques and examples of applications are debated to reveal the potential of the current state-of-the-art technology, and to further promote the usefulness of tandem MS.     

Mass spectrometry for small molecule pharmaceutical product development: a review

Developing a pharmaceutical product has become increasingly difficult and expensive. With an emphasis on developing project knowledge at an earlier stage in development, the use of information-rich technologies (particularly MS) has continued to expand throughout product development. Continued improvements in LC/MS technology have widened the scope of utilizing MS methods for performing both qualitative and quantitative applications within product development. This review describes a multi-tiered MS strategy designed to enhance and accelerate the identification and profiling of both process- and degradation-related impurities in either the active pharmaceutical ingredient (API) or formulated product. Such impurities can be formed either during chemical synthesis, formulation, or during storage. This review provides an overview of a variety of orthogonal-mass spectrometric methodologies, namely GC/MS, LC/MS, and ICP-MS, in support of product development. This review is not meant to be all inclusive; however, it has been written to highlight the increasing use of hyphenated MS techniques within the pharmaceutical development area.     

LC/MS applications in drug development

The combination of high-performance liquid chromatography and mass spectrometry (LC/MS) has had a significant impact on drug development over the past decade. Continual improvements in LC/MS interface technologies combined with powerful features for structure analysis, qualitative and quantitative, have resulted in a widened scope of application. These improvements coincided with breakthroughs in combinatorial chemistry, molecular biology, and an overall industry trend of accelerated development. New technologies have created a situation where the rate of sample generation far exceeds the rate of sample analysis. As a result, new paradigms for the analysis of drugs and related substances have been developed. The growth in LC/MS applications has been extensive, with retention time and molecular weight emerging as essential analytical features from drug target to product. LC/MS-based methodologies that involve automation, predictive or surrogate models, and open access systems have become a permanent fixture in the drug development landscape. An iterative cycle of "what is it?" and "how much is there?" continues to fuel the tremendous growth of LC/MS in the pharmaceutical industry. During this time, LC/MS has become widely accepted as an integral part of the drug development process. This review describes the utility of LC/MS techniques for accelerated drug development and provides a perspective on the significant changes in strategies for pharmaceutical analysis. Future applications of LC/MS technologies for accelerated drug development and emerging industry trends are also discussed.     

Mass spectrometry‐based holistic analytical approaches for metabolite profiling in systems biology studies

Metabonomics and metabolomics represent one of the three major platforms in systems biology. To perform metabolomics it is necessary to generate comprehensive “global” metabolite profiles from complex samples, for example, biological fluids or tissue extracts. Analytical technologies based on mass spectrometry (MS), and in particular on liquid chromatography–MS (LC–MS), have become a major tool providing a significant source of global metabolite profiling data. In the present review we describe and compare the utility of the different analytical strategies and technologies used for MS‐based metabolomics with a particular focus on LC–MS. Both the advantages offered by the technology and also the challenges and limitations that need to be addressed for the successful application of LC–MS in metabolite analysis are described. Data treatment and approaches resulting in the detection and identification of biomarkers are considered. Special emphasis is given to validation issues, instrument stability, and QA/quality control (QC) procedures. © 2011 Wiley Periodicals, Inc., Mass Spec Rev 30:884–906, 2011     

眼组织中缓释地塞米松药膜的HPLC/MS/MS测定方法研究

眼部手术后 ,在眼组织的前房中植入缓释地塞米松药膜以防术后感染。本实验采用液液萃取、液固萃取的方法来提取生物样品中的地塞米松药物 ,经液相色谱 /质谱 /质谱进行检测 ,样品中药物浓度在 1 0 8— 54μg/L范围内呈现良好的线性 (γ2 =0 .9995) ,最低检测浓度为 0 4 μg/L ,平均回收率大于 77%。用这一方法可高灵敏度、高选择性地应用于临床相关药物的动力学研究     

The role of liquid chromatography-mass spectrometry in the determination of heroin and related opioids in biological fluids.

The opioid most commonly sold in the illicit market is heroin. This substance, classified as an analgesic narcotic drug, has an extremely short half-life, and it is rapidly metabolized to 6-monoacetyl-morphine and further to morphine. Morphine is principally metabolized by conjugation to morphine-3 and morphine-6 glucuronides. Morphine itself is a potent analgesic that is frequently used in the pharmacological intervention of cancer pain. The toxicological and clinical evaluation of heroin and morphine have stimulated pharmacokinetic studies in human and animal models. Although a number of methods exist to determine opiates and their metabolites, liquid chromatography (LC) appears to be the technique that can separate without any pretreatment the lipophilic and the hydrophilic analytes of the complete metabolic profile of heroin and/or morphine. Moreover, mass spectrometry (MS) used as a detector for liquid chromatography is unique, because it offers universality and selectivity. Furthermore, efforts have been made to develop LC/MS interfaces that could overcome the previous problem of poor sensitivity. For this reason, in recent years LC combined with MS has been applied to the analysis of opiates--parent drugs and metabolites--in biological fluids. This article reviews the existing literature on the determination, using liquid chromatography coupled to mass spectrometry, of opiate metabolites found in different biological matrices after the administration of the parent compounds.     

大气压电离液质联机的应用

本文以我们几年来的实际工作为素材，对电喷雾（ESI）和大气压化学电离（APCI）这一近年发展起来的软电离质谱接口技术做了应用性介绍。文中列举了该技术用于药物代谢、蛋白质构象以及热不稳定化合物分析的几个实例，对一些典型化合物在离子源内经碰撞诱导解离（CID）形成的质谱开裂产物进行了解析，对流量匹配、脱溶剂的技术特点、离子在溶液中的预形成、溶剂的选择、仪器的噪声等实践中遇到的问题做了较为详细的讨论。     

液相色谱—质谱法（LC—MS）分析细胞色素C酶解肽谱及一级结构

本文采用液相色谱—质谱联用法（LC／MS）,对一种蛋白质—细胞色素C（Cyt．C）的胰蛋白酶酶解产物进行了质量肽谱分析,确定了各肽段在蛋白质中的位置。并用串联质谱（MS／MS）分析了大部分肽段的序列,均获得满意的结果。     

液相色谱-离子阱质谱和飞行时间质谱分析橄榄油中的特丁津除草剂

This application describes the use of LC/Ion Trap MS and LC/TOF for the identification of and quantization of terbutylazine in olive oil samples. The analysis by ion trap was achieved in MS/MS mode, monitoring the characteristic fragment ion at m/z 174. The identification by LC/TOF was accomplishment with accurate mass, along with the accurate mass of chlorine isotope cluster present in terbutylazine. The accuracy typically is better than 2 ppm. The method sensitivity, linearity precision accuracy matrix effect, and limit of detection were also studied.     

超高效液相色谱(UPLCTM)与质谱联用技术:-改善药残和代谢物分析的结果质量

介绍了最新的UPLC^TM技术及其与质谱联用对药残及代谢物分析。UPLC与质谱联用不仅获得高速、高分离度，而且显著地提高质谱检测的灵敏度。对于需要高灵敏度的药残及其痕量代谢物的分析，UPIC/MS/MS技术是当今最有效的工具之一。     

Optimization of the linear quantification range of an online trypsin digestion coupled liquid chromatography–tandem mass spectrometry (LC–MS/MS) platform

Tandem mass spectrometry (MS/MS)-based proteomic workflows with a bottom-up approach require enzymatic digestion of proteins to peptide analytes, usually by trypsin. Online coupling of trypsin digestion of proteins, using an immobilized enzyme reactor (IMER), with liquid chromatography (LC) and MS/MS is becoming a frequently used approach. However, finding IMER digestion conditions that allow quantitative analysis of multiple proteins with wide range of endogenous concentration requires optimization of multiple interactive parameters: digestion buffer flow rate, injection volume, sample dilution, and surfactant type/concentration. In this report, we present a design of experiment approach for the optimization of an integrated IMER-LC–MS/MS platform. With bovine serum albumin as a model protein, the digestion efficacy and digestion rate were monitored based on LC–MS/MS peak area count versus protein concentration regression. The optimal parameters were determined through multivariate surface response modeling and consideration of diffusion controlled immobilized enzyme kinetics. The results may provide guidance to other users for the development of quantitative IMER-LC–MS/MS methods for other proteins.     

Liquid chromatography-mass spectrometry for the quantitative bioanalysis of anticancer drugs

The monitoring of anticancer drugs in biological fluids and tissues is important during both pre-clinical and clinical development and often in routine clinical use. Traditionally, liquid chromatography (LC) in combination with ultraviolet (UV), fluorescence, or electrochemical detection is employed for this purpose. The successful hyphenation of LC and mass spectrometry (MS), however, has dramatically changed this. MS detection provides better sensitivity and selectivity than UV detection and, in addition, is applicable to a significantly larger group of compounds than fluorescence or electrochemical detection. Therefore, LC-MS has now become the method of first choice for the quantitative bioanalysis of many anticancer agents. There are still, however, a lot of new developments to be expected in this area, such as the introduction of more sensitive and robust mass spectrometers, high-throughput analyses, and further optimization of the coupled LC systems. Many articles have appeared in this field in recent years and are reviewed here. We conclude that LC-MS is an extremely powerful tool for the quantitative analysis of anticancer drugs in biological samples.     

UHPLC-LTQ Orbitrap MS快速鉴别无患子果皮中部分苷及苷元成分

建立了超高效液相色谱-线性离子阱/静电场轨道阱组合式高分辨质谱(UHPLC-LTQ Orbitrap MS)联用技术整合多种数据采集、挖掘策略快速分析无患子果皮中苷类化合物;总结了两类无患子皂苷标准品的质谱裂解规律及特征碎片离子,用于无患子果皮中苷类成分的快速筛查和鉴别;探索了母离子列表(parent ion list,PIL)-MS2、PIL-MS3和高能碰撞(high energy collision dissociation,HCD)技术,用于无患子果皮中母核结构相近化合物的精细区分和鉴别;最后通过智能化的In silico策略(Mass Frontier软件中的FISH和MS Tree Match技术)对无患子果皮中新的苷类成分进行高通量筛查,并用HCD碎裂模式验证其预测的可靠性。结果表明,在无患子果皮中共鉴别出67个苷类化合物,其中38个化合物被快速筛查和鉴别,18个母核结构相近的化合物得以精细区分,11个新成分被推测。该方法能够为无患子质量控制及药效学研究提供参考数据,有助于复杂中药成分的快速定性分析。     

Development of LC/MS Techniques for Comprehensive Metabolome Analysis

Metabolomics is a rapidly evolving field for studying biological systems and discovering potential disease biomarkers. For any metabolomics application, metabolome analysis with adequate sensitivity and specificity is essential in defining the metabolome. Ideally, all metabolites present in a biological system are qualitatively and quantitatively profiled. Unfortunately, due to technical limitations, only a fraction of metabolites are currently analyzed by using techniques such as NMR and mass spectrometry (MS). Due to limited metabolome coverage, many important metabolome networks and some subdue changes in the metabolome may not be revealed with current techniques. In this presentation, several technical issues related to the development of LC/MS for enabling metabolome analysis will be discussed. Because of great diversity of chemical and physical properties of metabolites, we have been developing an isotope labeling LC/MS workflow with a goal of improving the metabolome coverage in analyzing biological samples such as human biofluids and tissue samples. Several labeling chemistries will be described to provide isotope tags to the metabolites for sensitive detection and accurate quantification. LC methods including multi-dimensional separation to separate the labeled metabolites with high efficiency will be discussed. New protocols for MS analysis, metabolite identification and quantitative data processing will be presented.     

Application of mass spectrometry in the analysis of polybrominated diphenyl ethers

This review summarized the applications of mass spectrometric techniques for the analysis of the important flame retardants polybrominated diphenyl ethers (PBDEs) to understand the environmental sources, fate and toxicity of PBDEs that were briefly discussed to give a general idea for the need of analytical methodologies. Specific performance of various mass spectrometers hyphenated with, for example, gas chromatograph, liquid chromatograph, and inductively coupled plasma (GC/MS, LC/MS, and ICP/MS, respectively) for the analysis of PBDEs was compared with an objective to present the information on the evolution of MS techniques for determining PBDEs in environmental and human samples. GC/electron capture negative ionization quadrupole MS (GC/NCI qMS), GC/high resolution MS (GC/HRMS) and GC ion trap MS (GC/ITMS) are most commonly used MS techniques for the determination of PBDEs. New analytical technologies such as fast tandem GC/MS and LC/MS become available to improve analyses of higher PBDEs. The development and application of the tandem MS techniques have helped to understand environmental fate and transformations of PBDEs of which abiotic and biotic degradation of decaBDE is thought to be one major source of Br_1‐9BDEs present in the environment in addition to direct loading from commercial mixtures. MS‐based proteomics will offer an insight into the molecular mechanisms of toxicity and potential developmental and neurotoxicity of PBDEs. © 2009 Wiley Periodicals, Inc., Mass Spec Rev 29:737–775, 2010