赛默飞Q Exactive Plus LCMS 系统

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]。

本次实验基于优化的样品前处理、纳升液相系统与Orbitrap高分辨质谱，建立了迄今为止最高灵敏度的质谱HCPs检测方法，并利用内标蛋白进行准确的相对定量。由于我们没有查到NIS抗体标准品ELISA定量结果，所以未进行进一步对比质谱与ELISA的一致性。接下来将利用此方法对商品化的抗体药物进行HCPs的定性和相对定量，以及考察方法的重现性。

本实验采用Q Exactive高分辨质谱仪测定BlotGlycoTM O-Glycan样品制备试剂盒制备的2AB标记的Enbrel O糖链，建立了Q Ex-active分析糖蛋白O-糖链的样品前处理、色谱和质谱分析方法，为糖蛋白O-糖链的研发和生产检测提供快速、准确的分析平台。

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。

本标准规定了同步测定饲料中-激动剂沙丁胺醇、菜克多巴胺和盐酸克仑特罗的液相色谱质谱联用法(LC-MS)法。本标准适用于配合饲料、浓缩饲料和添加剂预混合饲料中沙丁胺醇、菜克多巴胺和盐酸克仑特罗的测定。本标准中沙丁胺醇、莱克多巴胺和盐酸克仑特罗的检测限均为0.01mg/kg,定量限均为0.05 mg/kg.

The high resolution, accurate mass Exactive mass spectrometer was shown to be sensitive in detecting <0.1 ppb melamine (0.5 pg on column) in neat standard, and response is linear from 0.1 to 100 ppb. The error for mass accuracy is <2 ppm with lock mass. Milk samples prepared by dilute-and-shoot showed severe ion suppression that was reduced with a simple 1-minute isocratic HILIC LC separation, after which a consistent response factor of 0.3-0.5 for each sample matrix was obtained for quantitation. Quantitation limits were less than 44, 65, and 110 ppb for coffee cream, infant formula and 3-in-1 instant coffee mix, respectively, exceeding the requirement of 250 ppb LOQ as stated by US FDA for infant formula.

The high resolution, accurate mass Exactive mass spectrometer was shown to be sensitive in detecting <0.1 ppb melamine (0.5 pg on column) in neat standard, and response is linear from 0.1 to 100 ppb. The error for mass accuracy is <2 ppm with lock mass. Milk samples prepared by dilute-and-shoot showed severe ion suppression that was reduced with a simple 1-minute isocratic HILIC LC separation, after which a consistent response factor of 0.3-0.5 for each sample matrix was obtained for quantitation. Quantitation limits were less than 44, 65, and 110 ppb for coffee cream, infant formula and 3-in-1 instant coffee mix, respectively, exceeding the requirement of 250 ppb LOQ as stated by US FDA for infant formula.