我国临床质谱检测系统及量值溯源的现状与发展

近年来质谱技术及其应用在我国发展十分迅速,在临床检验领域尤其是在临床生化检验方面的作用越来越明显,特别是液相层析-串联质谱法(LC-MS/MS)在各种生物样品中小分子化合物的定量检测及临床检验量值溯源方面应用越来越广泛。目前市场上已有诸多与不同类型质谱仪相配套的商品化检测试剂盒,为质谱技术的临床应用提供了全套解决方案。本文重点介绍质谱技术及其临床应用、我国商品化质谱分析仪与配套检测试剂及其质量评价方法,同时对该领域的量值溯源情况(包括参考方法和参考物质)进行简要总结。     

基于LC-MS/MS测定人体类固醇激素的研究进展

类固醇激素是一类具有环戊烷多氢菲结构的化合物,由细胞色素P450酶催化形成,在调节机体代谢、促进性器官发育等方面起着重要作用。临床上将类固醇激素水平作为肾上腺疾病、精神类疾病等的诊断指标。液相色谱-串联质谱(LC-MS/MS)联用技术因具有高灵敏度、高通量和高专属性的特点,已成为类固醇临床测定的首选方法。本文综述了LC-MS/MS在内源性类固醇激素测定中的应用,特别是在样品前处理、色谱条件和质谱条件优化等方面的研究进展,为临床上类固醇激素的诊断检测提供参考。     

赛默飞推出应用包以简化水中激素的检测与分析

<正>2012年11月30日,赛默飞世尔科技于近期推出用于简化水质分析过程的赛默飞激素应用包。作为一款专为使用UltiMate3000系列液相色谱与赛默飞TSQ系列三重串联四极杆质谱联用的LC-MS/MS系统开发的应用包,它为简化液相色谱/串联质谱法(LC-MS/MS)检验分析地下水和饮用水水样中的激素污染物提供了一个完整的检测平台。     

LC-MS/MS和荧光偏振法监测环孢素A血药浓度的相关性研究

建立液相色谱-质谱（LC-MS/MS）监测环孢素A(CsA)血药浓度的方法,并与荧光偏振法（TDx）监测结果比较,进行统计学分析,考察2种方法的相关性。LC-MS/MS方法测定CsA血药浓度平均为（99.9±62.7） μg/L,小于TDx法测定的浓度平均值（199.7±129.1） μg/L,2种监测方法测定结果相关系数r平均为0.929,相关性较好,LC-MS/MS法和TDx法自身配对比值平均为（51.6±9.3）%,因此推测LC-MS/MS法可用于CsA血药浓度监测的治疗窗。LC-MS/MS法测定的浓度为CsA原型药物浓度,能更准确的反映原型药物在体内的准确浓度,更适用于日常血药浓度监测工作。     

氯霉素和磺胺类药物的LC-MS/MS分析

由于磺胺类抗球虫药和氯霉素(CAP)残留危害极大,可引起人类的癌症、畸形、抗药性及某些中毒现象.美国、欧盟等已明文规定禁止在食品工业中使用该类药物.在目前所有的检测氯霉素残留的方法中,LC-MS/MS以其无须衍生化,样品的制备简便、灵敏度高等特点而成为氯霉素残留的主要测试方法.本工作采用VARIAN1200L LC-MS/MS测定氯霉素和磺胺二甲嘧啶,测定浓度分别低至0.1×10-9量级和1×10-9量级,仍有良好的线性响应.     

GPC-LC-MS/MS法测定大米中8种农药残留

建立了凝胶渗透色谱(GPC)-液相色谱串联质谱(LC-MS/MS)法测定大米中8种农药残留的分析方法。样品用乙腈超声提取,提取液经过凝胶渗透色谱除去大分子杂质后,供LC-MS/MS分析;添加浓度为0.2μg时,回收率为71.3%~111.1%,相对标准偏差为3.64%~12.76%,方法具有自动化程度高、净化效果好、精密度好,分析快速等优点,适用于大米中多农药残留的日常检测工作。     

LC/MS和LC-MS/MS定性检测保健品中非法添加的伐地那非

采用LC/MS和LC-MS/MS法同时检测保健品中非法添加的伐地那非。用乙酸乙酯提取,以10 mmol•L^-1的甲酸铵（pH 3.5）和乙腈为流动相,分别用Agilent Zorbax SB C₁₈（150 mm×2.1 mm×5 μm）和Agilent Zorbax SB C₁₈（100 mm×2.1 mm×3.5 μm）色谱柱分离,采用电喷雾离子源,正离子扫描方式进行分析检测。该方法简便、快捷、可靠,适用于保健品中非法添加伐地那非的常规检测。     

液相色谱-串联质谱法快速检测干血点样品中尿酸

建立了一种高通量液相色谱-串联质谱（LC-MS/MS）检测干血点(DBS)样品中尿酸(UA)的方法。采用自动液体操作平台对样品进行高通量自动化前处理,首先用含有UA-1,3-¹⁵N₂稳定同位素内标的Tris水溶液进行萃取,然后用含有0.1%甲酸、0.05%三氟乙酸的乙腈溶液沉淀蛋白质。处理后的样品经CN色谱柱分离,多反应监测(MRM)模式进行LC-MS/MS分析。结果表明,在DBS样品中,UA在7.8~1 000 μmol/L浓度范围内的线性关系良好(R²＝0.999);检出限为3.1 μmol/L (S/N=3);定量限为12.5 μmol/L(S/N=10);平均回收率为95%~101%;日内相对标准偏差(RSD)为4.2%~12%;日间RSD为5.3%~14%。以样品中UA检测结果的总体RSD不超过15%来考察样品稳定性,分别将样品在-20 ℃保持30天、在37 ℃保持7天、反复冻融5次,样品中UA检测结果总体RSD小于10%,表明样品稳定性良好。将该方法与传统生化分析方法相比较,并分析了204份血样,相关性较好(R²＝0.946)。此方法可为有限采血条件下UA的检测及UA相关疾病的大规模筛查提供新途径。     

呋喃类物质代谢物的LC-MS/MS分析

呋喃类抗生素是一种治疗牲畜疾病的药物.由于该类药物可能致癌,美国、澳大利亚、加拿大、日本、新加坡、欧盟等已明文规定禁止在食品工业中使用该类药物.目前的检测方法以使用LC-MS检测其代谢物为主,以此间接测试呋喃类物质.呋喃类物质包括呋喃唑酮、呋喃它酮、硝呋妥因、硝呋醛等,它们的代谢物分别为AOZ、AMOZ、AH、SC.本工作使用Varian 1200L LCMS/MS,以正离子模式,通过检测呋喃类物质的代谢物,以此检测痕量的呋喃类物质.     

大米及玉米中6种环己烯酮类除草剂的液相色-串联质谱测定

建立了大米和玉米中6种环己烯酮类除草剂吡喃草酮,禾草灭,噻草酮,苯草酮,稀禾定和烯草酮的高效液相色谱-串联质谱(LC-MS/MS)测定方法。样品制备后,采用乙腈进行提取,经C₁₈和Envi-Carb固相萃取柱净化处理后,采用LC-MS/MS电喷雾电离（ESI）,多反应监测（MRM）正离子模式检测,外标法定量。在1~100 μg•L^-1范围内,6种环己烯酮类除草剂的线性相关系数均大于0.999。在添加浓度5~20 ng•g^-1范围内,6种环己烯酮类除草剂的回收率在70.0%~97.9%,相对标准偏差（RSD）均在10％以内。     

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     

Direct-EI in LC-MS: towards a universal detector for small-molecule applications

This review article will give an up-to-date and exhaustive overview on the efficient use of electron ionization (EI) to couple liquid chromatography and mass spectrometry (LC-MS) with an innovative interface called Direct-EI. EI is based on the gas-phase ionization of the analytes, and it is suitable for many applications in a wide range of LC-amenable compounds. In addition, thanks to its operating principles, it prevents unwelcome matrix effects (ME). In fact, although atmospheric pressure ionization (API) methodologies have boosted the use of LC-MS, the related analytical methods are sometime affected by inaccurate quantitative results, due to unavoidable and unpredictable ME. In addition, API's soft ionization spectra always demand for costly and complex tandem mass spectrometry (MS/MS) instruments, which are essential to acquire an "information-rich" spectrum and to obtain accurate quantitative information. In EI a one-stage analyzer is sufficient for a qualitative investigation and MS/MS detection is only used to improve sensitivity and to cut chemical noise. The technology illustrated here provides a robust and straightforward access to classical, well-characterized EI data for a variety of LC applications, and readily interpretable spectra for a wide range of areas of research. The Direct-EI interface can represent the basis for a forthcoming universal LC-MS detector for small molecules.     

A review of clinical diagnostic applications of liquid chromatography-tandem mass spectrometry

Liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) technology is emerging as a complementary method to traditional methodology used for clinical applications. Enhanced specificity and high-throughput capabilities are providing significant benefits to clinical diagnostic laboratories conducting routine analyses. This technology is expected to expand rapidly as scientists focus on more complicated challenges that can be solved efficiently by adding LC/MS/MS to their arsenal of techniques.     

液相色谱-串联质谱法测定猪肝中痕量己烯雌酚、己烷雌酚和双烯雌酚

建立了液相色谱-串联质谱法测定猪肝中痕量己烯雌酚、己烷雌酚和双烯雌酚。以氟化铵为流动相添加剂,目标物响应值平均提高约10倍,向样品中加入1%（V/V）乙酸乙腈超声提取,提取液经分散固相萃取净化、水稀释后,采用Waters Acquity UPLC HSS T3色谱柱,以0.1 mmol/L氟化铵和乙腈为流动相,梯度洗脱分离,电喷雾电离源负离子模式和多反应监测模式检测,外标法定量。在0.4、0.8、4.0 μg/kg添加水平下,3种目标物的加标回收率为89.5%～109.4%,相对标准偏差（RSD）为2.2%～7.8%,方法定量限为0.4 μg/kg。本方法前处理过程无需浓缩,操作简便、灵敏度高、重现性好、环保节约,适用于猪肝中痕量己烯雌酚、己烷雌酚和双烯雌酚的定性与定量分析。     

液相色谱-串联质谱检测兽药残留中的基质效应研究进展

液相色谱-串联质谱技术作为强有力的分析工具已被广泛应用于多个领域,但其检测中的基质效应对测定结果准确性的影响需引起重视。本文综述了液相色谱-串联质谱分析测定中基质效应的来源、重要性、产生的可能机制、影响因素、评价及消除和（或）补偿基质效应的方法,重点概括了典型兽药残留检测中应对基质效应的方法进展。     

仪器对液相色谱-同位素稀释质谱测试结果的影响

以同位素稀释质谱法测定奶粉中氯霉素为例，比较了近几年商品化的API5000、Q-trap5500、Agilent 6490、Waters XEVO TQS 4种高效液相色谱-串联三重四极杆质谱仪（LC-MS/MS）的关键指标，并重点研究了由于离子源设计导致的样品基质中残留化合物对氯霉素及其氘代同位素试剂测量的基质效应差异。通过比较测量氯霉素的线性、灵敏度和精密度等指标，发现这4种仪器测量氯霉素的线性和检测灵敏度差异不大。在0.2~1.0 ng/g痕量浓度范围分析时，在低浓度点时，不同仪器上两种化合物的离子化差异较大，测量精密度在1.0%~9.8%之间；在高浓度时，仪器对目标物响应的影响较小。考察了各仪器对不同基质溶液所产生的基质效应，发现在所有仪器上被测物与其同位素稀释剂的响应均存在差异，且随浓度增大差异越来越明显；不同预处理方法下得到的样品基质溶液，在不同仪器可能出现抑制或增强两种相反的基质效应现象，且强度差异也较大。表明基质中不同类型的干扰物在不同类型的离子源上的离子化模式与机理可能存在较大差异，从而导致同位素试剂之间响应的差异，在准确测量中必须分别进行基质效应的验证与确认。     

液相色谱-串联质谱生物分析方法的基质效应和对策

液相色谱-串联质谱（LC-MS/MS）法具有高灵敏度、高选择性、高通量等特点,已经成为生物分析的主流方法,广泛应用于新药发现和开发过程中新化学实体及其代谢物的定量分析。然而,由于生物样品基质成分复杂,共洗脱物质会影响分析物的离子化,使分析物的质谱响应增加或降低,从而影响LC-MS/MS分析方法的准确度和精密度。一般而言,离子抑制较离子增强更为常见。因此,在建立LC-MS/MS法时,需要对离子抑制进行评估和校正,并采用不同的策略消除或减少基质效应的影响。本文综述了生物样品分析中基质效应的来源和评估方法,重点介绍了克服基质效应的策略。引起基质效应的物质包括磷脂、盐类、尿素、代谢物等内源性物质和赋形剂、抗凝剂、固定相释放物质及降解产物等外源性物质。目前基质效应的评价方法主要有提取后加入法和柱后灌注法。克服基质效应的策略主要有使用稳定同位素内标,将极性药物衍生化,沉淀蛋白后稀释上清液,优化样品预处理方法、色谱和质谱条件等。结合本课题组的应用实例,重点阐述了建立LC-MS/MS生物分析方法时,为减少基质效应遇到的困难及解决方法。     

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     

基于质谱的蛋白质绝对定量研究策略和建议

科学家已经研究获得了大量的蛋白质/多肽潜在生物标志物,这些生物标志物需要经过验证和确证才能进一步转化到临床应用.针对蛋白质/多肽的绝对定量研究在标志物验证和确证过程中起到关键作用.传统的蛋白质定量方法,如酶联免疫吸附试验(ELISA)技术存在蛋白质抗体难以获得、不同抗体批次之间存在差异、基于抗体的检测存在交叉反应等问题...     

Mass spectrometry in sports drug testing: Structure characterization and analytical assays

Owing to the sensitive, selective, and unambiguous nature of mass spectrometric analyses, chromatographic techniques interfaced to various kinds of mass spectrometers have become the most frequently employed strategy in the fight against doping. To obtain utmost confidence in analytical assays, mass spectrometric characterization of target analytes and typical dissociation pathways have been utilized as basis for the development of reliable and robust screening as well as confirmation procedures. Methods for qualitative and/or quantitative determinations of prohibited low and high molecular weight drugs have been established in doping control laboratories preferably employing gas or liquid chromatography combined with electron, chemical, or atmospheric pressure ionization followed by analyses using quadrupole, ion trap, linear ion trap, or hyphenated techniques. The versatility of modern mass spectrometers enable specific as well as comprehensive measurements allowing sports drug testing laboratories to determine the misuse of therapeutics such as anabolic-androgenic steroids, stimulants, masking agents or so-called designer drugs in athletes' blood or urine specimens, and a selection of recent developments is summarized in this review.