Trace level detection of microbiological contaminants in pharmaceutical environment using Rapid Evaporative Ionisation Mass Spectrometry

使用快速蒸发电离质谱法检测制药环境中的痕量微生物污染物

• 批准号: 2092186
• 金额: --
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2092186
• 关键词: Trace; level; detection; microbiological; contaminants

Rapid evaporative ionization mass spectrometry (REIMS) yields highly specific phospholipid profiles of different bacteria and moulds which can be used to distinguish between different microorganisms. REIMS offers a new opportunity for the development of a lipid based, sample-preparation-free microbial identification system. This technique would be considerably faster than current methods and have the sensitivity and robustness compared to other techniques (e.g. MALDI).This project is focusing on developing technique to detect and categorise different microorganisms in the presence of drug substances, excipients in a tablet formulation and in liquids at a suitable detection limit. Another focus of the project is to develop a technique which would be used for identifying isolates from environmental monitoring plates which would be an enabler for future real time release (as part of a wider control strategy). Pharmaceutical water analysis, absence/presence of specified organisms clauses which are required for a range of pharmaceutical dosage presentations are another two possible REIMS applications which this project is focusing. Comparing with existing methods and procedures in pharmaceutical companies, REIMS technology could potentially be a technique with great savings for the business (e.g. saving time and money, increasing sustainability, integrity and robustness, reducing delays for batch and stock release).Below, I have put together project plan for the first 2 years and introduced timelines to show which part of the plan have been started or will be started in the upcoming months.Big part of the project is environmental monitoring. In order to develop a technique which would be able to identify the plates, construction of pharmaceutical environmental isolates data base is required. Hundreds of samples (bacteria and moulds) are send to Imperial everyweek from AstraZeneca, where samples are saved in to the sample data base for further use and identification by MALDI or 16S sequencing (about 80% of samples have to be sequenced to obtain true identification which is crucial for further technique development due to the not matching results from AZ used techniques for identification and Imperial MALDI).1st -2nd Year Work Plan:1. Construction of environmental REIMS identification databasea. Comparison between environmental and clinical samplesb. Construction and validation of REIMS identification database c. Comparison to AZ (AstraZeneca) in-house techniquesd. Samples analysis using REIMS, reference model building and validatione. Investigation of samples storage conditions effect for analysis on REIMS2. AZ water samples analysis on REIMS (purified water, water for injections, borehole water)a. Develop technique to concentrate and enrich samples for analysisb. Identify lower limit detection/CFU of bacteria required by regulationsc. Identify method for E.coli and Coliform detection such as short incubation, isotopic labelling, magnetic separation or biomarkers analysis.d. Identify method for viable bacteria detection (reference model building forprediction approach)3. Fungi Identificationa. Construction and validation of REIMS identification databaseb. Investigation of fungi colonies metabolic changes during 7 day growth onREIMSc. Samples analysis using REIMS, reference model building and validation d. Investigation of samples storage conditions effect for analysis on REIMS4. Culturing free approach for drug substances or drug itselfa) Investigate solvents suitable for dissolving drug substances if requiredb) Develop technique to concentrate samples for analysis (filtering approach)c) Lower limit detection/CFU of bacteria required by regulationsd) Identify method for viable cell detection such as short incubation, isotopic labelling, magnetic separation or biomarkers analysis.5. Instrument developmenta. Investiga

快速蒸发电离质谱法（REIM）产生了不同细菌和霉菌的高度特异性磷脂谱，可用于区分不同的微生物。 Reims为开发基于脂质的，预先准备的微生物识别系统提供了新的机会。与其他技术相比，该技术将比当前方法快得多，并且具有敏感性和鲁棒性（例如Maldi）。该项目集中于开发技术，以检测和分类在存在药物的情况下，在片剂配方和摄取中的药物，并分类不同的微生物。在适当的检测极限的液体中。该项目的另一个重点是开发一种技术，该技术将用于从环境监测板中识别隔离板，这将是未来实时释放的推动者（作为更广泛的控制策略的一部分）。药物水分析，一系列药物剂量演示所需的指定生物条款的缺失/存在是该项目集中的另一种可能的Reims应用。与制药公司的现有方法和程序进行比较，Reims技术可能是一种为业务节省大量的技术（例如，节省时间和金钱，增加可持续性，完整性和鲁棒性，减少批处理和股票释放的延迟）。制定项目计划的最初两年，并介绍了时间表，以显示该计划的哪一部分已经启动或将在即将到来的月份开始。该项目的BIG部分是环境监控。为了开发能够识别板的技术，需要制造药物环境隔离株数据库。数百个样本（细菌和模具）从阿斯利康发送到帝国每周，在那里将样品保存到样本数据库中，以通过MALDI或16S测序进行进一步使用和识别（必须测序约80％的样品以获得真实的鉴定这对于进一步的技术开发至关重要，因为AZ使用的技术与识别和帝国MALDI不匹配的结果。构建环境Reims标识数据库。环境和临床样品之间的比较。 REIMS标识数据库的构建和验证c。与AZ（Astrazeneca）内部技术的比较。使用Reims，参考模型构建和验证的样品分析。对样品储存条件的研究效果，以分析Reims2。亚利桑那州水样在奖励上进行了分析（纯净水，注射水，钻孔水）a。开发浓缩和丰富样品的技术以进行分析。确定法规要求的细菌的下极限检测/CFU。识别大肠杆菌和大肠菌群检测的方法，例如短孵育，同位素标记，磁分离或生物标志物分析。确定可行细菌检测的方法（参考模型构建供应方法）3。真菌识别。 REIMS标识数据库的构建和验证。对真菌菌落在7天生长期间的代谢变化的研究。使用Reims，参考模型构建和验证d的样品分析d。对样品储存条件的研究效果，以分析Reims4。培养药物或药物本身的免费方法）研究适合于溶解药物的溶剂（如果需要）b）开发技术以浓缩样品进行分析（滤波方法）c）c）通过法规所需的细菌的下限检测/CFU，确定可行细胞检测的方法例如短孵育，同位素标记，磁分离或生物标志物分析5。仪器开发。调查