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The 4th Open Symposium of the European Bioanalytical Forum entitled ‘Less is More’ was held on 16–18 November 2011 at the Hesperia Tower Hotel, Barcelona, Spain. More than 50 interesting presentations were delivered covering areas with interest for the small- and large-molecule community – biomarker validation; regulations, including an update on new and emerging guidelines and on Global harmonization; technology updates; incurred sample stability; microdosing; dried blood spots and microsampling; challenges of ‘free’ and ‘total’ macromolecule quantification; stability issues in ligand binding assays or anomalous results. In excess of 450 delegates from more than 170 institutes and companies (industry, regulators and academia) from all global regions participated in the open and stimulating discussions. This manuscript provides an overview of the highlights discussed at the meeting.

Background: Dried blood spot (DBS) technology was evaluated for the quantitative determination of guanfacine in human blood in clinical studies. A very sensitive DBS assay has been developed using HPLC coupled with an AB Sciex 5500 QTRAP® (Applied Biosystems/MDS Sciex, ON, Canada) MS system (LC–MS/MS) with a linear calibration range of 0.05 to 25 ng/ml. High-resolution MS using an Exactive Orbitrap® (ThermoFisher, LLC., CA, USA) was compared with the QTRAP using extracted exact mass ion current profiles for guanfacine and its stable-isotope-incorporated internal standard. The sample preparation employed liquid–liquid extraction with methyl t-butyl ether of 5 mm punched DBS card disks, followed by reversed-phase HPLC separation coupled with either MS/MS or high-resolution MS. Routine experiments were performed to establish the robustness of the DBS assay, including precision, accuracy, linearity, selectivity, sensitivity and long-term stability of up to 76 days. In addition, several factors that potentially affect quantitation were investigated, including blood volume for DBS spotting, punch size and punch location. Results: A sensitive research assay with a LLOQ of 0.05 ng/ml was developed and subjected to several components of a method validation common to a regulated bioanalysis procedure employing DBS. This method development and partial validation study determined that spot volume, punch size or punch location do not affect assay accuracy and precision. The DBS approach was successfully applied to a clinical study (a Phase I, randomized, double-blind, placebo-controlled, crossover study to assess the effect of varying multiple oral doses of guanfacine on the pharmacokinetic, pharmacodynamic, safety, and tolerability profiles in healthy adult subjects). The pharmacokinetic profiles for 12 volunteers generated from the DBS assay and from a previously validated plasma assay were compared and were found to be comparable. DBS incurred samples collected from finger prick blood and directly applied to the DBS cards were also analyzed for comparison. Conclusion: From a bioanalytical perspective, DBS sample collection and analysis is a potentially viable alternative for guanfacine determination in clinical studies, utilizing approximately 100 µl of blood per subject profile compared with a few millilitres of blood drawn for conventional plasma bioanalysis.

The European Bioanalysis Forum is a bioanalytical nonprofit organization comprised of European pharmaceutical companies (27 members to date) and currently expanding to include CROs as well. The European Bioanalysis Forum provides a broad European bioanalytical network for the discussion of scientific, technological and regulatory topics of bioanalytical interest. The 3rd Annual Open Symposium was again much anticipated after the two previous successful meetings. The symposium included sessions on thinking outside the ‘commodity’ box, bioanalytical challenges with blood, global harmonization, assay platforms, dried blood spots, immunogenicity, matrix effects, anomalous results, biomarkers and two plenary technology sessions hosted by the Platinum sponsors. Experts and key opinion leaders were invited as guest speakers. A total of 424 delegates registered from 113 companies representing a large percentage of the European bioanalytical community. In addition to 48 oral presentations, 88 posters were presented and there was a vendor exposition of 40 companies.

The European Bioanalysis Forum is a non-profit organization comprised of European pharmaceutical companies (25 members to date). Their activities focus on bringing together managers and scientists in the broad field of bioanalysis to discuss topics related to science, process and regulations. There has been much interest over the past few years in the potential application of dried blood spots as an alternative to traditional plasma collection in pharmacokinetic studies. The success of the technique has been highlighted by several companies. We know that seven of the European Bioanalysis Forum member companies are using dried blood spots intensively and that 22 out of 25 companies are using it or plan to use it very soon, initially in nonregulated studies. However, most companies have less than 1 year of experience with dried blood spots and, beyond the scientific merit, it is less clear just how the technique is perceived by key client groups, such as toxicology, clinical and regulatory authorities. The objective of this symposium was to bring together representatives from all of these client groups as well as a broad bioanalytical audience to discuss the various perspectives on dried blood spots and to try to provide answers to potential issues that still remain. The symposium included sessions on dried blood spots in the nonregulated environment, toxicology and regulatory/QA perspectives, clinical use and bioanalytical applications and tools. There was plenty of time for discussion within the program in tutorials, poster and break-out sessions and the degree of delegate participation reflected the high level of engagement with the topic. A total of 190 delegates attended from more than 80 organizations. In addition to 21 oral presentations, more than 20 posters were presented and there was a vendor exposition of ten sponsor companies.

The European Bioanalysis Forum is a bioanalytical discussion group comprised of European pharmaceutical companies (27 members to date). The membership share a common vision to advance the shared understanding of topical concerns through discussion of scientific, technological and regulatory issues of bioanalytical interest. The 2nd Annual Open Symposium was much anticipated after the success of the first one. The symposium included sessions on the European Medicines Agency draft bioanalytical guidance, challenges in the analysis and validation of biomarkers, metabolite quantification issues, dried blood spot technology, the analysis of proteins and peptides by MS, latest hot topics in ligand-binding assays, unresolved issues in method validation and a plenary session from the Platinum sponsors. Experts and key opinion leaders were invited as guest speakers. A total of 420 delegates (up from 251 delegates in 2008) attended from 160 organizations, representing a large percentage of the European and international bioanalytical community. In addition to 40 oral presentations, 72 posters were presented and there was a vendor exposition of 35 companies.

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.