赛默飞 Vanquish Flex UHPLC系统

研究表明，根据 EPA 方法要求，采用 TSQ Quantis 三重四极杆MS 定量分析饮用水中的藻毒素和节球藻素具有灵敏度、准确度、重现性和可靠性。进样 5 μL 可获得足够的灵敏度。

Show the straightforward method development capabilities of the Thermo Scientific™ Vanquish™ UHPLC platform in combination with Thermo Scientific™ Chromeleon™ 7.2 Chromatography Data System (CDS) software.

To demonstrate the simple transfer of an analytical HPLC method from an Agilent 1260 Infinity system to a Vanquish Flex UHPLC system.

Analytical method transfer and method modernization can be a barrier to upgrading to the latest technologies. Revalidating an existing method while continuing to meet regulatory guidance can be a challenging and time-consuming endeavor. However, method modernization is often less difficult than the common perception, and good practices can be put in place to streamline and facilitate the process. This case study will address some of the concerns related to method transfer and provide guidance from a leading contract development and manufacturing organization (CDMO) authority. A method transfer guide onto the Thermo Scientific™ Vanquish™ UHPLC platform (CS000566)1 based on these guiding principles complements this document.

ChromSword Chromeleon Connect, Vanquish Flex, mebendazole, impurity analysis, UHPLC, automated method development, robustness test.

随着治疗性生物产品在全球药物市场所占份额的不断提高，各国监管机构通过发布质量源于设计（Quality by Design, QbD）原则等手段，对生物药的质量控制提出了更高的要求。对于生物制药企业而言，在生产及批次放行过程中对关键质量属性（critical quality attribute, CQA）和杂质等进行鉴定、定量和监控就显得尤为重要。传统的质控手段包括多种分离手段，如反相色谱（reversed-phase high performance liquid chromatography, RP-HPLC)、体积排阻色谱（size-exclusion chromatography, SEC）、离子交换色谱（ion-exchange chromatography, IEX）和毛细管电泳（capillary electrophoresis, CE）等。 2015年，Rogers等基于Orbitrap高分辨质谱平台发展了Multi-Attribute Method（MAM），用于在一针进样中同时对CQA进行鉴定、定量和实时监控[1]。自从MAM发布以来，获得了业界的极大关注，在近年的学术会议中屡屡成为热点议题[2]。

本文基于Vanquish Flex生物兼容超高效液相色谱平台搭建了 Trap column的二维液相（2D-LC），无需拆除已搭建好的管 路，直接通过软件进行Simple Switch切换可将二维液相切换成 并联液相（Dual LC）；首先用Vanquish Dual LC系统加速方法 开发，获得较为理想的一维杂质分离方法。然后基于一维方法 建立RP-RP模式的二维液相方法对卡贝缩宫素中的杂质进行MS 定性分析。Simple Switch是市面上唯一的解决方案，可极大地 提高该套配置仪器的灵活性、便捷性和实用性，为用户增加投 资回报。

The biopharmaceutical industry continues to develop mAb-based biotherapeutics in increasing numbers. Due to the complexity of these biotherapeutics, there are several key quality attributes (CQAs) that need to be measured and controlled to guarantee their safety and efficacy. The presence of aggregates in a formulated drug product must be assessed to avoid potential issues with immunogenicity.

Key Words Glycan Quantitation, Glycan Profiling, Glycoprotein, Native Glycan, Universal Detection, Mixed-mode Separation Goal To develop fast and sensitive UHPLC methods suitable to directly measure the glycan content and profile of glycoproteins.

Key Words SMART Digest kit, SOLAμ, Vanquish, peptides, protein, bottom up proteomics, proteomics, trypsin digestion, buffer, mAb, biomolecules, biotherapeutics, biopharmaceutical, biocompatible UHPLC, Acclaim C18 RSLC, peptide mapping, in-solution digestion, UHPLC, accurate mass, QE, Orbitrap, peptide quantitation, SPE, solid-phase extraction, Q Exactive Plus

Key Words SMART Digest kit, SOLAμ, Vanquish, peptides, protein, bottom up proteomics, proteomics, trypsin digestion, buffer, mAb, biomolecules, biotherapeutics, biopharmaceutical, biocompatible UHPLC, Acclaim C18 RSLC, peptide mapping, in-solution digestion, UHPLC, accurate mass, QE, Orbitrap, peptide quantitation, SPE, solid-phase extraction, Q Exactive Plus

Key Words Glycan Quantitation, Glycan Profiling, Glycoprotein, Native Glycan, Universal Detection, Mixed-mode Separation Goal To develop fast and sensitive UHPLC methods suitable to directly measure the glycan content and profile of glycoproteins.

The comprehensive information acquired by vDIA mode generated more efficient screening and confirmation results while the result of targeted quantitation was also gained at the same time, which suggested good performance for both qualitative and quantitative analysis with vDIA mode.

The comprehensive information acquired by vDIA mode generated more efficient screening and confirmation results while the result of targeted quantitation was also gained at the same time, which suggested good performance for both qualitative and quantitative analysis with vDIA mode.

6-苄基腺嘌呤( 6 - benzylaminopurine, 6-BA)是一种人工合成的细胞分裂素，与植物内源激素具有相似的结构和性质，能够抑制植物叶内叶绿素、核酸和蛋白质分解，广泛用作无根豆芽的生长调节剂。

本文尝试采用比ELSD灵敏度更高的CAD检测器，及柱性能更好的核壳型色谱柱对白酒中三氯蔗糖进行检测，取得了较为理想的效果：方法定量限可达0.04 mg/kg，满足GB22255-2014定量限7.5mg/kg的基本要求。

本实验利用可变数据非依赖采集（vDIA），建立了一种兽药分析的新方法。该法具备高灵敏度和高选择性，能够获得被测样品全面而高质量的数据，并可将定量分析与非目标物和未知物筛查相结合。

建立了亲水作用鄄反相二维液相色谱鄄串联质谱测定乳制品中茁鄄内酰胺、四环素、大环内酯、氨基糖苷、酰胺醇、喹诺酮和磺胺7 类20 种抗生素残留的方法。样品与C18 和CN 填料混合,进行基质固相分散萃取,以乙腈和水洗脱,洗脱液旋转蒸发至近干,残渣流动相溶解后分析。对样品前处理条件、色谱流动相、质谱参数进行了优化。各分析物回归方程的相关系数为0. 9945 ~0. 9998;以定量离子信噪比为3 和10 时所对应的样品中分析物浓度计算检出限和定量限,分别为0. 10 ~ 2. 40 滋g/ kg 和0. 33 ~ 7. 92 滋g/ kg。奶粉和牛奶中添加水平25 滋g/ kg 的加标回收率分别为72. 5% ~97. 2%和70. 1% ~96. 8%,相对标准偏差为4. 2% ~8. 8%和3. 7% ~9. 9%。本方法应用于实际样品测定,结果满意。

近年来，以瘦肉精为代表的兽药残留已逐渐成为人们普遍关注的一个社会热点问题。药物残留不仅可以直接对人体产生急慢性毒性作用，引起细菌耐药性的增加，还可以通过环境和食物链的作用间接对人体健康造成潜在危害。动物源食品中兽药残留的检测具有待测物质浓度低，样品基质复杂，干扰物质多、动物种类多样，对药物代谢存在差异等问题，一直以来是食品安全检测中的难点。因此为保证人们健康和食品安全，必须建立一套快速、有效、覆盖面广的动物源食品中兽药残留的筛查方法。

本实验利用可变数据非依赖采集（vDIA），建立了一种兽药分析的新方法。该法具备高灵敏度和高选择性，能够获得被测样品全面而高质量的数据，并可将定量分析与非目标物和未知物筛查相结合。

TSQ Altis 三重四极杆质谱仪为血浆中的雌二醇和睾酮的临床定量分析研究提供了卓越的灵敏度。在该篇技术文摘所描述的方法中，当采用该方法测定血浆中的睾酮和雌二醇时，定量限别为 20 pg/mL（柱上进样 200 fg）和 2 pg/mL（柱上进样 20 fg）。

本文建立了一种基于Vanquish超高效液相色谱和QE Focus超高分辨质谱在血清中检测类胰岛素生长因子的方法。该方法采用两步蛋白沉淀和在线固相萃取的方法来实现样品的净化，最终获得了满足高通量临床实验室日常检测的结果。该方法分析每个样本的时间为5分钟，但实际上目标和内标化合物出峰的时间窗口不足20秒，因此在后续的工作中我们可以采用Transcend II TLX-4四通道在线前处理液相系统来将实验室的检测通量提升4倍， 即一分半钟测试一个样本。

A Vanquish Flex Quaternary UHPLC system was applied for the SEC analysis. This is a low dispersion, inert UHPLC, which can be used successfully for this type of application. For this analysis, pre-column dispersion was intentionally introduced to show the effects of dispersion in front of the column at different flow rates.

利用Orbitrap超高分辨质谱得到高精度的一级质谱信息来分析完整胰岛素，从而绕开胰岛素不容易产生碎片离子的问题。此外，对于胰岛素这一类大肽段类分析物，合适的色谱分析条件也尤为重要，我们使用ProSwift RP-4H整体柱获得了极佳的分析效果。

分析方法在整个分析的动态范围内呈现出 优异的线性和重现性。该方法表明，TSQ Altis 质谱提供了药物 化合物分析中所需的灵敏度和重现性。

建立了一种灵敏、快速的测定辛伐他汀的 LC-MS/ MS 方法，经方法学验证本方法的专属性、精密度和准 确度、基质效应、回收率和稳定性等符合生物样品分析 法规要求，适用于药物生物等效性一致性评价研究。