Peter van Amsterdam

Incurred sample reproducibility: views and recommendations by the European Bioanalysis Forum

Following intensive discussions, review, alignment of procedures and multiple surveys among their member companies, the European Bioanalysis Forum (EBF) is providing a recommendation on how to integrate incurred sample reproducibility (ISR) in the bioanalytical process. The recommendation aims to provide guidance throughout the lifecycle of a validated method, including the application of the method in study support. In its recommendation, the EBF considers both the internal discussions with EBF member companies, as well as the input provided in international meetings where ISR was discussed. The ultimate goal of the EBF recommendation is to ensure that bioanalytical methods can provide accurate and reproducible concentration data for pharmacokinetic and/or toxicokinetic evaluation, without any compromise, while safeguarding the optimal use of laboratory resources.

The European Bioanalysis Forum community’s evaluation, interpretation and implementation of the European Medicines Agency guideline on Bioanalytical Method Validation

The European Medicines Agencys (EMA) 2011 guideline on bioanalytical method validation (BMV) was evaluated and subsequently intensely discussed by the European Bioanalysis Forum (EBF) during a 2-day workshop (EBF Workshop on the implementation of the EMA guideline on BMV, Château de Limelette, Limelette, Belgium, 15-16 March 2012). The goal of the evaluation and discussions was to come to a uniform interpretation of the guideline and thus to help facilitate a smooth implementation at our laboratories. Up front preparations for the workshop by dedicated teams concentrated on challenges on implementation: ambiguities, technical or operational challenges and issues in general. In addition, common understandings were identified as well as main differences to the 2011 US FDA guideline. The guideline was perceived as being well written with a clear structure, separating method validation from sample analysis and treating all relevant aspects one-by-one in a logical order. It is the first BMV guideline clearly addressing the specifics for ligand binding assays and it shows a good match with current scientific thinking. The EBF community considers the EMA BMV guideline an excellent basis for countries that are in the process of developing or updating their own BMV guideline.

Anticoagulant counter ion impact on bioanalytical LC-MS/MS assays: results from discussions and experiments within the European Bioanalysis Forum.

BACKGROUND In regulated bioanalysis, the need for partial validation when changing the counter ion of the anticoagulant is currently being debated within the bioanalytical community. To date, industry and the health authorities have not yet reached a consensus on this issue. The aim of the present study was to evaluate the impact of a change in counter ion when using the same anticoagulant on LC-MS/MS assay performance for a broad array of new chemical entities, compiling data generated at companies within the European Bioanalysis Forum (EBF). RESULTS In all, 15 EBF member companies provided experimental data on partial validation. In total, data from 42 LC-MS/MS assays were evaluated. The results show that a change in counter ion when using the same anticoagulant had no impact on assay performance. CONCLUSION Based on these results and on conclusions from previous studies, the EBF recommends that in regulated bioanalysis, plasma samples containing different counter ions, but the same anticoagulant, should be regarded as equal matrices, thus removing any need for partial validation.

Stabilization of clinical samples collected for quantitative bioanalysis – a reflection from the European Bioanalysis Forum

In bioanalysis of small molecules, the analyte concentration in the measured samples should reflect the concentration during sample collection. Precautions may be needed to prevent over- or under-estimation of the obtained result. This might require the addition of stabilizers to prevent degradation or nonspecific binding. For unstable drugs, it is essential to know how analytes can be stabilized before the start of the clinical study. Although the stabilization methods are well documented, the impact of the stabilization on the clinical workflow is not properly addressed. Already during method development, the clinical implications in terms of personnel safety, ease of use, training possibilities and staff capacity should be taken into account, and validation of the bioanalytical method should reflect collection procedures.

Introduction to the Proposals from the Global Bioanalysis Consortium Harmonization Team

Bioanalysis in Pharmaceutical Research & Development has been at the crossroad of science, technology, and regulations from its onset [1]. The first American Association of Pharmaceutical Scientists (AAPS)/Food and Drug Administration (FDA) Bioanalytical Workshop in 1990 in Crystal City, USA (CCI) was a major landmark for the industry to agree on execution and documentation of bioanalytical experiments. And although the resulting conference report was not a regulatory guidance, it was a roadmap for bioanalysts around the world for a decade on how to validate and apply bioanalytical methods [2]. In early 1999, the FDA promoted bioanalysis in support of pharmacokinetic evaluation in clinical and preclinical studies more formally into the regulated space by issuing a draft guidance. After broad industry consultation during an AAPS/FDA meeting again in Crystal City (CC-II) in early 2000 [3], FDA issued a guidance for Industry on Bioanalytical Method Validation in May 2001 [4]. This guidance provided the bioanalytical community the regulatory framework for bioanalytical method validation and the application of these methods. Although the guidance set clear expectations in many areas, the lack of detail on expectations in some paragraphs resulted in different interpretations in industry or by the inspectors. This, in combination with the rapid technological advancements in separation sciences, mass spectrometry, and ligand binding assay (LBA) or cell-based assay formats, resulted in individual views on how to validate bioanalytical methods and apply them to routine sample analysis. In trying to provide assistance in the interpretation of this guidance, the FDA continued to reach out to industry at conferences. In addition, some inspection findings documented in Form 483s provided a way forward. The 2001 guidance only referred to LBAs that enabled small molecule analysis and did not address the bioanalysis of macromolecules. In support of the rapidly growing ligand binding assay scientific community, two additional white papers were published to cover LBA aspects of macromolecule bioanalysis [5, 6]. To further discuss and clarify the 2001 FDA Guidance, FDA/AAPS organized a next meeting in Crystal City (CC-III), from which a comprehensive conference report was issued [7]. Crystal City III also put Incurred Sample Reanalysis (ISR) back on the foreground, a controversial topic which was further discussed during Crystal City IV [8]. Next, as mentioned in the Kudoh paper [1], multiple other guidelines or best practices on GLPs, GCPs, computer/software, or analytical instrument validation increasingly impacted the bioanalytical laboratory. Last but not least, other regions/countries besides the USA issued their own guidelines on regulated bioanalysis [9–11]. Building the Global Bioanalytical Consortium At the initiative of four regional professional organizations from Europe (European Bioanalysis Forum (EBF)) and North America (AAPS, Applied Pharmaceutical Analysis (APA) APA and Calibration & validation group (CVG—currently represented in GBC by Canadian Forum for Analytical and Bioanalytical Sciences CFABS), a letter was sent to the FDA and the European Medicine Agency (EMA)), formally requesting the health authorities and bioanalytical community to join hands and harmonize global bioanalysis scientific best practices. The need for global harmonization of the bioanalytical guidance was also supported by publishing this letter as an open letter [12], an initiative which was supported by many [13–15]. At the same time, the authors from aforementioned open letter together with additional representatives from these organizations (currently referred to as founding members), proposed to form an organization which brings together experts from the global bioanalytical community to discuss, share, and finally propose a harmonized view on bioanalytical best practices that could lead to a guidance: the Global Bioanalysis Consortium (GBC). A consensus was reached among around 280 delegates, including five Regulatory Agencies, during the 5th Workshop on Recent Issues in Bioanalysis (5th WRIB) in April 2010 on the main characteristic of what a “harmonization and Global Bioanalytical Guidance” should be based upon the following: science driven with inclusion of a rationale behind each requirement to prevent “box checking”. In addition, it should have a global perspective (not local issues), should not be prescriptive, and finally must get buy-in from all the countries [16]. From there, the GBC founding members proposed the mission of the organization [17] and reached out into the global bioanalytical community to build the GBC ensuring balanced representation from North America, Latin America, Europe/Middle East/Africa and Asia-Pacific.

Use of polarity switching for the simultaneous bioanalysis of analytes with three orders of magnitude difference in concentration by LC–MS/MS

BACKGROUND The challenge of quantifying two compounds in a single assay with drastic dynamic ranges is to obtain linearity without source or detector saturation at the mass spectrometer. RESULTS In positive-ionization mode, the nonlinear relationships for Desmethyl Mebeverine Acid (DMAC) were demonstrated using three common strategies to overcome this issue: using offset voltage parameters, less-sensitive product ion or 13C mass SRM transitions. On the contrary, nonlinear relationships for DMAC were overcome if negative-ionization mode was used. Due to Mebeverine analytical LLOQ, dilution was not suitable for a single assay of Mebeverine and DMAC. However, polarity switching in negative mode for DMAC was successfully found to compensate for the nonlinearity at the mass spectrometer while preserving Mebeverine linear regression model in positive mode. CONCLUSION The polarity switching strategy has demonstrated the advantage of improving linearity for analytes having different ionization polarities and three orders of magnitude difference in concentration.

Conference Report: AAPS and US FDA Crystal City V meeting on Quantitative Bioanalytical Method Validation and Implementation: feedback from the EBF

Crystal City V meeting on Quantitative Bioanalytical Method Validation and Implementation: 2013 Revised US FDA Guidance 3-5 December 2013, Hilton Baltimore, MD, USA The meeting provided an opportunity for Industry and regulators from the US FDA to discuss the recently published revised draft FDA Guidance for Industry on Bioanalytical Methods Validation during the 90 day review period. Key perspective and philosophical positions were shared leading to a healthy exchange of views and ideas on topics in the revised document. Discussions covered all aspects of bioanalytical method validation and method utilization. However, the main dialogue was focused on chromatographic methods, ligand-binding assay methods and biomarker analysis. The resulting open debate led to greater understanding of the document, but also provided clear feedback, including the request on harmonization with approved Bioanalytical Methods Validation guidances release from other health authorities, as well as the consensus view between industry and the FDA. Members of the European Bioanalysis Forum summarized prospective discussions during the meeting in Baltimore; however, this Report is not intended to constitute the official proceedings from the meeting, which are expected to be published later this year.

Conference Report: The 3rd EBF Focus Meeting: ‘Hatching’ – emerging technologies approaching the regulated bioanalysis laboratory

One hundred and eighty scientists from industry and academia discussed the progress in emerging technologies approaching regulated bioanalysis, with a focus on what potential hurdles prevent them becoming broadly accepted by industry and regulators. The conference delegates agreed that moving innovative technologies forward can only be achieved by providing solid data to support the application. In addition, also establishing an open dialogue with health authorities is key for success. By successfully integrating new technologies in the bioanalytical laboratory we contribute to bringing more safe and efficacious therapies faster to patients.