The 18th GCC Closed Forum was held in New Orleans, LA, USA, on 11 April 2025. Representatives from international bioanalytical Contract Research Organizations were in attendance to discuss scientific and regulatory issues specific to bioanalysis. The issues discussed at the meeting included: Compliance with US FDA Guidance for Biomarkers; Emerging Technologies; Sample Reconciliation Efforts; Chicken anti-Human IgG antibody; Post-extraction Sample Stability; Singlicate for LBA; Discrepancies of ELISpot data; Regulatory Inspection Experiences; Cross-validation Harmonization in Regulated Bioanalysis; and ELN/LIMS. Conclusions and consensus from discussions of these topics are included in this article.

The 17th GCC Closed Forum was held in San Antonio, TX, USA, on 10th May 2024. Representatives from international bioanalytical Contract Research Organizations were in attendance in order to discuss scientific and regulatory issues specific to bioanalysis. The topics discussed at the meeting included integrative bioanalysis, patient-centric sampling, emerging technologies, data integrity, sample reconciliation efforts, discrepancies of ELISpot data, cross-validation harmonization, ultrasensitive platforms for immunogenicity assays, remote regulatory assessments, shedding assays by qPCR/dPCR, and biomarker assays. Conclusions and consensus from discussions of these topics are included in this article.

Biomarker analysis enables a deep understanding of physiological and biological processes and offers insights into pathological disease states and conditions. When measured in tissues, the spatial distribution of biomarkers may be evaluated. To meet regulatory and sponsor requirements, guidance on the approach to validation and the parameters to be evaluated is essential. The main goals of this GCC white paper are to disseminate the survey results discussed during the 16th& 17thGCC Closed Forums (2023 & 2024) and to provide recommendations from the GCC members on technical and regulatory considerations for the bioanalysis of biomarkers in tissues.

Gene therapy, cell therapy and vaccine research have led to an increased need to perform cellular immunity testing in a regulated environment to ensure the safety and efficacy of these treatments. The most common method for the measurement of cellular immunity has been Enzyme-Linked Immunospot assays. However, there is a lack of regulatory guidance available discussing the recommendations for developing and validating these types of assays. Hence, the Global CRO Council has issued this white paper to provide a consensus on the different validation parameters required to support Enzyme-Linked Immunospot assays and a harmonized and consistent approach to Enzyme-Linked Immunospot validation among contract research organizations.

Gene therapy, cell therapy and vaccine research have led to an increased use of qPCR/ddPCR in bioanalytical laboratories. CROs are progressively undertaking the development and validation of qPCR and ddPCR assays. Currently, however, there is limited regulatory guidance for the use of qPCR and a complete lack of any regulatory guidelines for the use of the newer ddPCR to support regulated bioanalysis. Hence, the Global CRO Council in Bioanalysis (GCC) has issued this White Paper to provide; 1) a consensus on the different validation parameters required to support qPCR/ddPCR assays; 2) a harmonized approach to their validation and 3) a consistent development of standard operating procedures (SOPs) for all the bioanalytical laboratories using these techniques.

The 13th Global CRO Council (GCC) closed forum for bioanalysis was held in New Orleans, LA, USA on 5 April 2019. This GCC meeting was organized to discuss the contents of the 2019 ICH M10 Bioanalytical Method Validation Draft Guideline published in February 2019 and consolidate the feedback of the GCC members. While ICH M10 will cover requirements for reference standards, one of the biggest challenges facing the CRO community is the lack of consistency and completeness of Certificates of Analysis for reference standards used in regulated bioanalysis. Similar challenges exist with critical reagents (e.g., capture and detection antibodies) used for assays supporting biologics. The recommendations provided in this publication are the minimum requirements for the content that GCC members believe should be included in Certificates of Analysis for reference standards obtained from commercial vendors, sponsors and compendial suppliers, for use in regulated bioanalytical studies. In addition, recommendations for internal standards, metabolites and critical reagents are discussed.

The 13th GCC Closed Forum for Bioanalysis was held in New Orleans, Louisiana, USA on April 5th, 2019. This GCC meeting was organized to discuss the contents of the 2019 ICH M10 Bioanalytical Method Validation Draft Guideline published in February 2019 and consolidate the feedback of the GCC members. In attendance were 63 senior-level participants from eight countries representing 44 bioanalytical CRO companies/sites. This event represented a unique opportunity for CRO bioanalytical experts to share their opinions and concerns regarding the ICH M10 Bioanalytical Method Validation Draft Guideline and to build unified comments to be provided to the ICH.

The 12th GCC Closed Forum was held in Philadelphia, PA, USA, on 9 April 2018. Representatives from international bioanalytical Contract Research Organizations were in attendance in order to discuss scientific and regulatory issues specific to bioanalysis. The issues discussed at the meeting included: critical reagents; oligonucleotides; certificates of analysis; method transfer; high resolution mass spectrometry; flow cytometry; recent regulatory findings and case studies involving stability and nonclinical immunogenicity. Conclusions and consensus from discussions of these topics are included in this article.

Over the last decade, the use of biomarker data has become integral to drug development. Biomarkers are not only utilized for internal decision-making by sponsors; they are increasingly utilized to make critical decisions for drug safety and efficacy. As the regulatory agencies are routinely making decisions based on biomarker data, there has been significant scrutiny on the validation of biomarker methods. Contract research organizations regularly use commercially available immunoassay kits to validate biomarker methods. However, adaptation of such kits in a regulated environment presents significant challenges and was one of the key topics discussed during the 12th Global Contract Research Organization Council for Bioanalysis (GCC) meeting. This White Paper reports the GCC members’ opinion on the challenges facing the industry and the GCC recommendations on the classification of commercial kits that can be a win-win for commercial kit vendors and end users.

The 11th Global CRO Council Closed Forum was held in Universal City, CA, USA on 3 April 2017. Representatives from international CRO members offering bioanalytical services were in attendance in order to discuss scientific and regulatory issues specific to bioanalysis. The second CRO-Pharma Scientific Interchange Meeting was held on 7 April 2017, which included Pharma representatives' sharing perspectives on the topics discussed earlier in the week with the CRO members. The issues discussed at the meetings included cumulative stability evaluations, matrix stability evaluations, the 2016 US FDA Immunogenicity Guidance and recent and unexpected FDA Form 483s on immunogenicity assays, the bioanalytical laboratory's role in writing PK sample collection instructions, biosimilars, CRO perspectives on the use of chiral versus achiral methods, hybrid LBA/LCMS assays, applications of fit-for-purpose validation and, at the Global CRO Council Closed Forum only, the status and trend of current regulated bioanalytical practice in China under CFDA's new BMV policy. Conclusions from discussions of these topics at both meetings are included in this report.

The 9th GCCClosed Forum was held just prior to the 2015 Workshop on Recent Issues in Bioanalysis (WRIB) in Miami, FL, USA on 13 April 2015. In attendance were 58 senior-level participants, from eight countries, representing 38 CRO companies offering bioanalytical services. The objective of this meeting was for CRO bioanalytical representatives to meet and discuss scientific and regulatory issues specific to bioanalysis. The issues selected at this year's closed forum include CAPA, biosimilars, preclinical method validation, endogenous biomarkers, whole blood stability, and ELNs. A summary of the industry's best practices and the conclusions from the discussion of these topics is included in this meeting report.

This was a single dose mass balance and metabolite characterization study of the antimalarial agent pyronaridine. Six healthy male adults were administered a single oral dose of 720 mg pyronaridine tetraphosphate with 800 nCi of radiolabeled (14)C-pyronaridine. Urine and feces were continuously collected through 168 h post-dose, with intermittent 48 h collection periods thereafter through 2064 h post-dose. Drug recovery was computed for analyzed samples and interpolated for intervening time periods in which collection did not occur. Blood samples were obtained to evaluate the pharmacokinetics of total radioactivity and of the parent compound. Total radioactivity in urine, feces, and blood samples was determined by accelerator mass spectrometry (AMS); parent concentrations in blood were determined with LC/MS. Metabolite identification based on blood, urine, and feces samples was conducted using a combination of LC + AMS for identifying radiopeaks, followed by LC/MS/MS for identity confirmation/elucidation. The mean cumulative drug recovery in the urine and feces was 23.7 and 47.8 %, respectively, with an average total recovery of 71.5 %. Total radioactivity was slowly eliminated from blood, with a mean half-life of 33.5 days, substantially longer than the mean parent compound half-life of 5.03 days. Total radioactivity remained detectable in urine and feces collected in the final sampling period, suggesting ongoing elimination. Nine primary and four secondary metabolites of pyronaridine were identified. This study revealed that pyronaridine and its metabolites are eliminated by both the urinary and fecal routes over an extended period of time, and that multiple, varied pathways characterize pyronaridine metabolism.

[目的]建立凤仙花药材中槲皮素、山奈酚的提取方法,对凤仙花药材中槲皮素、山奈酚进行薄层鉴别.[方法]采用羧甲基纤维素钠硅胶G板分离凤仙花药材中槲皮素、山奈酚成分.[结果]供试品色谱在与对照品色谱相应位置上显相同颜色的斑点,可用于鉴别凤仙花中的槲皮素和山奈酚.[结论]薄层色谱方法简便、专属性强、重现性好,可用于凤仙花药材的鉴别.

The topic of incurred sample stability (ISS) has generated considerable discussion within the bioanalytical community in recent years. The subject was an integral part of the seventh annual Workshop on Recent Issues in Bioanalysis (WRIB) held in Long Beach, CA, USA, in April 2013, and at the Global CRO Council for Bioanalysis (GCC) meeting preceding it. Discussion at both events focused on the use of incurred samples for ISS purposes in light of results from a recent GCC survey completed by member companies. This paper reports the consensus resulting from these discussions and serves as a useful reference for depicting ISS issues and concerns, summarizing the GCC survey results and providing helpful recommendations on ISS in the context of bioanalytical method development and application.

BioanalysisVol. 6, No. 20 CommentaryFree Access7th GCC Insights: incurred samples use; fit-for-purpose validation, solution stability, electronic laboratory notebook and hyperlipidemic matrix testingMario Rocci, Steve Lowes, Ronald Shoup, Fabio Garofolo, Raymond Farmen, Tianyi Zhang, John Allinson, Dominique Gouty, Roger Hayes, Robert Nicholson, Richard Houghton, Isabelle Dumont, Richard LeLacheur, Jennifer Zimmer, Maria Cruz Caturla, Philippe Couerbe, Kayode Awaiye, Saadya Fatmi, Curtis Sheldon, Joseph Bower, Michelle Fiscella, Douglas Fast, Stephanie Cape, Jim Hulse, John Kamerud, Stephanie Pasas-Farmer, Wei Garofolo, Marc Moussallie, Mike Buonarati, Nadine Boudreau, Brigitte Pellerin, Jenny Lin, Allan Xu, Mohammed Bouhajib, Mary Stipancic, Corey Nehls, Mark Warren, Shane Karnik, Craig Stovold, Scott Reuschel, Laura Cojocaru, John Marcelletti, Xinping Fang, Ian Smith & Andrea WatsonMario Rocci ICON Laboratories, Whitesboro, NY, USA, Steve Lowes Quintiles Bioanalytical and ADME Labs, Ithaca, NY, USA, Ronald Shoup AIT Bioscience, Indianapolis, IN, USA, Fabio Garofolo Algorithme Pharma, Laval, Quebec, Canada, Raymond Farmen Celerion, Lincoln, NE, USA, Tianyi Zhang Frontage Laboratories, Shanghai, China, John Allinson Previously with ICON Laboratories, Manchester, UK - Currently with LGC, Fordham, UK, Dominique Gouty Previously with Intertek, San Diego, CA, USA - Currently with BioAgilytix, Durham, NC, USA, Roger Hayes MPI Research, Mattawan, MI, USA, Robert Nicholson PPD, Richmond, VA, USA, Richard Houghton Quotient Bioresearch, Fordham, UK - Presently LGC, Isabelle Dumont Algorithme Pharma, Laval, Quebec, Canada, Richard LeLacheur Agilux Laboratories, Worcester, MA, USA, Jennifer Zimmer Alturas Analytics, Moscow, ID, USA, Maria Cruz Caturla Anapharm Europe, Barcelona, Spain, Philippe Couerbe Atlanbio, Saint-Nazaire, France, Kayode Awaiye BioPharma Services, Toronto, Ontario, Canada, Saadya Fatmi Biotrial Bioanalytical Services, Laval, Quebec, Canada, Curtis Sheldon Celerion, Lincoln, NE, USA, Joseph Bower Covance Laboratories, Chantilly, VA, USA, Michelle Fiscella Covance Laboratories, Chantilly, VA, USA, Douglas Fast Covance Laboratories, Madison, WI, USA, Stephanie Cape Covance Laboratories, Madison, WI, USA, Jim Hulse EMD Millipore, St. Charles, MO, USA - Presently Eurofins Pharma Bioanalytics Services US, John Kamerud EMD Millipore, St. Charles, MO, USA - Presently Eurofins Pharma Bioanalytics Services US, Stephanie Pasas-Farmer Frontage Laboratories, Exton, PA, USA, Wei Garofolo Global CRO Council for Bioanalysis (GCC), Marc Moussallie Previously with HLS, Somerset, NJ, USA, Mike Buonarati Intertek, El Dorado Hills, CA, USA, Nadine Boudreau InVentiv Health Clinical, Quebec City, Quebec, Canada, Brigitte Pellerin InVentiv Health Clinical, Quebec City, Quebec, Canada, Jenny Lin JCL Bioassay USA, Hoffman Estates, IL, USA, Allan Xu Keystone Bioanalytical, North Wales, PA, USA, Mohammed Bouhajib Pharma Medica Research, Mississauga, Ontario, Canada, Mary Stipancic Pharma Medica Research, Mississauga, Ontario, Canada, Corey Nehls PPD, Middleton, WI, USA, Mark Warren Previously with PRA International, Lenexa, KS, USA - Currently with Lambda Therapeutic Research, Toronto, Ontario, Canada, Shane Karnik Pyxant Labs, Colorado Springs, CO, USA, Craig Stovold Quotient Bioresearch, Fordham, UK - Presently LGC, Scott Reuschel Tandem Labs, Salt Lake City, UT, USA, Laura Cojocaru Tandem Labs, West Trenton, NJ, USA, John Marcelletti Tandem Labs, San Diego, CA, USA, Xinping Fang XenoBiotic Laboratories, Plainsboro, NJ, USA, Ian Smith York Bioanalytical Solutions, York, UK & Andrea Watson York Bioanalytical Solutions, York, UKPublished Online:21 Nov 2014https://doi.org/10.4155/bio.14.231AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInReddit Keywords: biomarker validationfit-for-purpose validationelectronic laboratory notebookELNhyperlipidemic matrixincurred sample stabilityincurred samplesISSlipemic plasmasolution stabilityThe Global CRO Council for Bioanalysis (GCC), a global independent consortium bringing together many senior level CRO representatives, was created in 2010 in order to openly discuss and share opinions on scientific and regulatory issues specific to the bioanalytical field [1]. Since its formation, the GCC members have met on a regular basis to discuss various topics and challenges faced by bioanalytical CRO companies. Several conference reports of past GCC meetings were published to share the discussions held during these events [2–4]. In addition, the GCC also published several influential White Papers on topics of widespread interest in bioanalysis [5–9]. These White Papers provide unified GCC recommendations that are helpful to the global bioanalytical community.The 7th GCC Closed Forum for Bioanalysis took place in Long Beach, CA, USA on 8 April 2013, one day before the start of the 7th Workshop on Recent Issues in Bioanalysis (WRIB). In attendance were 46 senior-level participants from six countries, representing 37 bioanalytical CRO companies/sites. This event represented a good opportunity for bioanalytical experts from CROs to share and discuss these issues of concern for the outsourcing industry.The 7th GCC Closed Forum was chaired by Mario Rocci, who started the meeting by communicating the official admonition statement, as has been done in all previous editions [1]. Prior to initiating discussions, participants introduced themselves. As in previous GCC meetings, multiple topics of current interest in bioanalysis were on the agenda of this closed forum. The nine topics for this 7th GCC meeting were the following:• Use of incurred samples for metabolite testing and method specificity assessments during method development;• Issues in performing incurred sample stability (ISS);• Fit-for-purpose method validation;• Determination of the stability of stock and working solutions in relation to the EMA guideline;• Electronic laboratory notebooks (ELN);• Discussion of recent US FDA findings;• Hyperlipidemic matrix testing;• Outcome of the GCC survey regarding sponsor requests versus regulatory expectations;• Feedback on the GCC and EBF recommendations on the validation of biomarkers.Use of incurred samples for metabolite testing & method specificity assessments during method developmentThe use of incurred samples collected as part of a trial for purposes other than initial analysis and various re-assays (analytical repeats, dilution repeats and incurred sample reanalysis [ISR]) can be of value when developing bioanalytical assays. However, regulatory and ethical concerns exist in using samples beyond their intended use as specified in the study protocol. In order to gauge the views of the bioanalytical CRO community on the benefits versus the risks of using incurred samples for method development purposes, a survey was generated and circulated to GCC member companies. In addition to metabolite testing and method specificity assessments, this survey included other purposes for the use of incurred samples, such as assisting in the determination of sample collection requirements, assay calibration ranges and quality control (QC) placement, selection of appropriate internal standards (IS), provisional stability, differential testing between LC–MS and LBA methods, as well as assessment of endogenous levels in biomarker assays.The results of this survey, which were presented during the meeting, expressed the view that there are obvious advantages to performing additional experiments with incurred samples as part of the development of robust LC–MS and LBA analytical methods for the generation of accurate and reliable data. In some situations, information obtained from experiments using these samples is critical to understanding method parameters. For instance, methods for bioequivalence (BE) studies would benefit from conducting tests on incurred samples only in some specific cases (e.g., unavailability of reference standard for metabolites), whereas for first-in-human studies, aspects such as testing for metabolites are more pertinent. It was also recognized that the information obtained from the use of incurred samples is often critical in establishing immunogenicity and biomarker assays.Sound practices, such as the use of pooled samples with adequate documentation, were suggested to minimize the potential for incurred samples misuse. In addition and as necessary, appropriate consent of volunteers/patients should be pursued to eliminate ethical concerns. During the discussion, it was acknowledged that the benefits and the risks associated with the use of incurred samples should be evaluated on a case-by-case basis considering the type of assay, its intended use, the resources involved and the value of the information that could be acquired. It was also clear that this benefit-to-risk approach must always take into account the importance of generating quality data when applying bioanalytical methods as part of sample analysis studies.This topic was also discussed as part of the 7th WRIB; a summary of the discussions and subsequent recommendations are presented in the 2013 White Paper in Bioanalysis [10].Issues in performing ISSWhile not a regulatory requirement, the conduct of ISS has been a topic of considerable interest in recent years and was extensively discussed as part of the 2012 and 2013 WRIB White Papers in Bioanalysis [10,11]. A portion of the GCC survey on the use of incurred samples was specifically dedicated to ISS questions, and the results of these survey questions were presented and discussed during the 7th GCC meeting. From the survey results, it was clear that most participants do not routinely conduct ISS experiments, and are in favor of performing ISS testing only in specific situations. Many respondents believe that incurred samples should be used to support ISR as part of additional ISR failure investigation experiments, and most respondents feel that ISS should not be included in regulations, but rather only a recommendation for use on a case-by-case basis. In addition to ISR failure investigations, examples of situations where ISS testing is warranted included back-conversion of unknown conjugates to analytes of interest, unstable glucuronides and other metabolites, stability in presence of co-administered compounds in oncology studies, inability to support stability with QC samples, enzymatic degradation of analytes and major matrix instability.Further discussion of this issue focused on the type of samples collected (blood, tissues, etc.), the sample collection/processing procedures employed and the determination of the 'time zero' concentration. Despite these potentially confounding issues, the value of investigating the stability of incurred samples was recognized under appropriate circumstances. An indirect way to assess ISS can be through the ISR evaluation, where incurred samples are re-assayed after a certain time interval has passed. While it can be valuable to perform ISR testing early to identify potential issues of concern as soon as possible, performing ISR testing later in the study can provide useful insights on the stability of incurred samples. Due to the importance of this topic, GCC decided to share its recommendations on ISS with the global bioanalytical community. A GCC White Paper on ISS was published in the September 2014 issue of Bioanalysis (Future Science Ltd, London, UK) [12].Fit-for-purpose method validationThe concept of fit-for-purpose method validation, which refers to an assay that does not fully meet current regulatory specifications for method validation, is another topic that has been discussed thoroughly at bioanalytical meetings in recent years. Interesting views and recommendations have been presented in several publications [2,9–10,13–16]. In non-regulated areas, the application of a fit-for-purpose approach is commonly applied to clinical therapeutics and biomarker studies, especially when encountering challenging assays with limitations. Fit-for-purpose validations, however, can pose challenges when applied in a highly regulated environment where the methods are driven by established regulatory guidance requirements.GCC members responded to several survey questions related to fit-for-purpose validation and associated method and validation parameters. Although there is concern that the use of a fit-for-purpose approach may have the potential to be negatively received by regulators if not applied correctly, the vast majority of survey respondents have not received regulatory deficiency findings when applying the fit-for-purpose approach. This could suggest that either the data were simply not submitted to regulatory authorities or the approach is being applied through the use of scientifically sound science, accompanied by documented justification that the approach employed was suitable for the intended purpose of the assay.Even though fit-for-purpose is often applied for particularly challenging types of assays, it was agreed that it should not be applied solely because of difficulties faced with a given assay. Rather the approach should be reserved for demonstrating acceptable assay performance when alternatives are not available.During the GCC meeting, the attendees were strongly in favor of a scientific and well-implemented application of a fit-for-purpose strategy, considering the end use of the bioanalytical data obtained. In other words, the quality of a quantitative assay should match the quality or importance of the decision being made based on the data generated.Determination of the stability of stock & working solutions in relation to the EMA guidelineThe stability of the analyte and IS in solution was a topic of discussion during this GCC Closed Forum. Given that solution preparation is time-consuming and costly, consideration should be given to best practices for prolonging the use of stock and working solutions while maintaining robust science.The current EMA Guideline on bioanalytical method validation (BMV) states the following regarding solution stability: "Stability of the stock and working solutions should be tested with an appropriate dilution, taking into consideration the linearity and measuring range of the detector" [17]. It is generally agreed that the stability evaluations in solvent are considered distinct and independent for a given type of solvent. When solutions of multiple concentrations are prepared for a given solvent, a bracketing approach is typically applied, where stability is assessed at the highest and lowest concentrations only. With this approach, the shortest stability duration available for the specified solvent is applied to solutions falling within the bracketed concentration range. Another important parameter that was considered during the meeting was the container material in which the solution is stored, which should be taken into account when assessing stability.Other aspects of solution stability discussed at the meeting were related to the acceptance criteria and the application of expiry dates. The maximum percent difference acceptance criterion applied between stability and comparison solutions appears to vary among CROs. Such criterion is not present in the FDA and EMA guidance documents. Some attendees expressed discomfort with the concept of 'daisy-chaining', which refers to expiry dating sub-stock solutions without regard to the age of the stock solution used in the preparation of the sub-stock. No clear consensus was achieved by the attendees on this 'age old' issue. Other discussion revolved around the impact of using a stock solution that was extensively used on previous occasions (retrieved many times from its storage location).In most cases, the expiry date of a solution is set based on the stability established for such a solution through testing. This does not systematically imply that the solution is not stable for a longer period (unless this has been established through testing). Consequently, the term 'retest date' was suggested as a more representative term than 'expiration date' for solutions where no instability has yet been demonstrated.ELNTraditionally, paper-based approaches have been used at bioanalytical laboratories for the documentation and recording of experiment conduct, standard operating procedures (SOPs),forms and other various documents. The emergence of ELN as an alternative to paper documentation is relatively recent in bioanalytical laboratories. Some organizations performing bioanalysis have decided to move forward in the implementation of this approach in their continuous search for quality improvement and better regulatory compliance.In order to seek the views and current state of ELN use at CROs, survey questions specific to ELN were sent to GCC members. The results of this survey indicate that paper continues to be the most widely used format for the vast majority of respondents and that ELN implementation is limited. This is surprising, considering the fact that several years have passed since ELN systems were first developed and proposed to bioanalytical facilities. ELN can be employed for many diverse applications, such as real-time recording of manual raw data, management of SOPs and other documents, collection of instrument raw data, calculation of secondary results (derived data), training and avoiding the inadvertent use of inadequate or uncalibrated equipment/materials and so on.Although ELN can be applied to a wide range of documentation tasks, the main reason CROs provided for not implementing an ELN system is the anticipated high cost of purchase and implementation of the system. Additional obstacles highlighted include the regulatory risk posed by novel systems that were not extensively tested yet; the difficulty in finding appropriate staff resources and time to implement, configure, customize and maintain the system; and the significant disruption to current operations.Some respondents currently using ELN acknowledge the advantages associated with its use, such as improved quality and completeness of records, as well as regulatory compliance. On the other hand, ELN users did not see much positive impact on their productivity, and the flexibility to handle different types of contracts was found to be an issue. Thus, the CRO community appears to be divided in considering ELN as the preferred choice for data and document recording and management.Discussion of recent FDA findingsThis GCC Closed Forum provided the opportunity to discuss recent FDA citations received by CROs. More specifically, the FDA findings received as part of a first FDA inspection conducted for a clinical bioequivalence study in China were shared. One legitimate question raised was whether these findings will be covered in the new FDA Draft Guidance on BMV [18], which had not yet been issued at that time? The first finding of particular interest was the failure to present long-term stability data in the presence of co-administered compounds. Such a finding was also previously observed as part of FDA inspections. Considerable debate and effort were recently expressed by the bioanalytical industry regarding this issue, as reported in the GCC White Paper specifically dedicated to this topic, as well as in the 2010, 2011 and 2012 White Papers in Bioanalysis [8,11,15–16]. The recommendation presented in the GCC White Paper is clear: a systematic evaluation of this stability is not believed to be required when scientific justifications suggest the absence of stability issues caused by the presence of the co-administered compound. The question remains as to whether the FDA will continue to cite such findings when this issue is not specifically included in the new 2013 draft BMV FDA guidance.Another thought-provoking FDA observation received for the study conducted in China pertained to IS response variability during sample analysis. Different approaches for IS response criteria were discussed in the past as part of the 2nd GCC Closed Forum [5]: the first approach being to establish upper and lower boundaries for IS response that would trigger a response or action should the criteria fail to be met, and the second approach consisting of a trend analysis of the IS variation of known samples to define the range of acceptability for IS variation in unknown samples. Pros and cons exist in applying either approach, as further explained in the GCC publication. It is generally agreed that acceptable performance of an assay can still be demonstrated despite the presence of IS variability. Suggested evidence that could be used to establish adequate performance may include acceptable ISR results, good accuracy of QCs prepared in pre-dosed subject matrix and low variability in relative response factors. The importance of having clear predefined criteria in SOPs and being ready to scientifically defend observations involving IS variability was also reiterated during the meeting.Other FDA comments, not listed as findings, were also shared at the meeting; they related to the conduct of partial versus full validation at a CRO laboratory when the method had previously undergone a full validation by the sponsor or another CRO, and the conduct of long-term stability testing at a CRO laboratory (i.e., not using existing stability data from the client or another CRO in the original full validation).A recurring question raised regarding findings or observations from regulatory agencies in general relates to whether the findings reflect the opinion of the individual inspector or of the agency as a whole? The global bioanalytical industry can significantly benefit from a certain level of uniformity within and between agencies in the application of regulatory requirements.Hyperlipidemic matrix testingThe need to test hyperlipidemic matrix during BMV was first introduced by ANVISA back in 2003, and is now included in current EMA and ANVISA guidelines as part of the matrix effect (EMA), and for both selectivity and matrix effect (ANVISA) [17,19]. While the EMA guideline is not particularly prescriptive on the procedural requirements for hyperlipidemic matrix testing, the ANVISA guideline provides slightly more details and defines a lipemic sample as "a high lipids degree sample, for example, coming from postprandial collection". This topic was brought to the industry's attention in the past year and was also part of the agenda of the 2013 7th WRIB, where issues were discussed and recommendations proposed [10].The value of including hyperlipidemic matrix testing during validation was deemed justified by the attendees, considering the likelihood that laboratories will handle and analyze study samples containing a high level of lipids and the potential effects of the presence of lipids on bioanalytical method performance. It was mentioned that the context of studies should be taken into account (e.g., high-fat fed studies, type of subjects enrolled, etc.), and the conduct of this test needs to be primarily science-driven. As a general approach, in order to be scientifically meaningful, the test should be representative of the samples to be analyzed and, thus, should take into account the approximate lipemia levels in matrix expected in study samples. Another issue discussed was the type of lipids to consider in the selection of an adequate matrix to be used for the test. Should specific classes of lipids be evaluated, such as triglycerides, phospholipids, or total cholesterol? With the goal of being representative, the use of 'artificial' lipemic matrix (i.e., matrix mixed with fat emulsion) may not represent a desirable approach, as opposed to the use of 'natural' matrix from high-fat donors (from subjects with high triglycerides levels or from subjects following a high-fat meal).Some concerns were also debated regarding the information obtained from vendors of lipemic matrix, which may pose problems as the information provided on the matrix lots is often insufficient. For instance, is the matrix supplied natural or artificial? If artificial, what is the identity and quantity of the fat emulsion used? If natural, from what type of donor? If from postprandial collection, what was the meal composition and collection time? It was concluded that several aspects need to be considered and many questions still remain unanswered regarding hyperlipidemic matrix testing during method validation.Outcome of the GCC survey regarding sponsor requests versus regulatory expectationsIt is generally understood that the global bioanalytical community is operating in a challenging environment with similar but different regulatory guidances and their interpretations, varying levels of experience within regulatory agencies, sponsors and CROs, timeline pressures, the need to reduce costs and increasing regulatory expectations. In an effort to eliminate some hurdles and improve workflows, initiatives were taken by both the industry and the regulatory agencies toward global harmonization, and specific groups were formed, such as the GCC in 2010 and the Global Bioanalysis Consortium (GBC) in 2011.A survey was sent to the GCC member companies where various sponsor requests were presented that could put the CRO at risk of facing regulatory issues and thus being 'caught in the middle' between sponsor requests and regulatory expectations. As CROs are frequently inspected and, thus, often have fresh regulatory insights, this survey served to gather useful opinions on such situations. As part of the survey, the respondents ranked the frequency of each sponsor request and the perceived regulatory risk associated with it. The responses were grouped by their risk factor based on a combination of frequency of a particular request and the perceived risk associated with the request. Each topic was then grouped as: 1 – relatively low overall concern; 2 – relatively moderate overall concern; 3 – relatively high overall concern. For example, whereas a particular sponsor request may carry a high level of regulatory objection, if the frequency of the request was rare, that specific scenario received a ranking of low general risk to the CRO community. Those scenarios categorized as high risk were associated with both a regulatory concern as well as a relatively high level of request frequency.For the survey results, the sponsor requests showing low general concern included:• To change the status from GLP to non-GLP;• To follow Sponsor's SOPs as opposed to CRO procedures/criteria;• To use a correction factor to normalize immunoassay results when standards do not match;• To add clarifications to archived documents;• To run Bioequivalence/Bioavailability (BE/BA)study by period to accelerate timelines;• To use Sponsor's spreadsheets that prevent verification of calculations;• To select ISR samples without a priori criteria for selection.The requests posing moderate concern were:• To use a Sponsor's decision tree for reporting re-assay results that omits considering data from acceptable runs;• To not follow CRO criteria for items not addressed in a Sponsor's SOP when a Sponsor's SOP is followed;• To report data from failed runs if the QCs in the range of the study samples are acceptable;• To employ contract language implying Sponsor's insertion between CRO and study director for GLP studies;• To use the analytical assay for different regulatory purposes than validated;• To transfer an assay validated elsewhere and only perform precision and accuracy testing as partial validation to support the transfer;• To provide input to the bioanalytical report that draws conclusions outside the scope of the data or does not reflect a conclusion reached by the CRO;• To not include ISRs in a regulated study (clinical study or first method application in preclinical study);• To omit valid results for various reasons.Finally, there were seven sponsor request scenarios where high overall concern was expressed (frequent occurrence combined with a high regulatory risk). They are listed below:• Short sample analysis timelines for a large number of samples, leading to a high percentage of initial and ISR results being generated prior to potential problems being identified (e.g., ISR failure, placement of QCs relative to unknown samples, etc.);• Re-assay of selected samples (PK repeats) without objective a priori criteria for selection;• Claims of GLP compliance for non-GLP clinical work;• Use of long-term stability data generated by the Sponsor or third party without details regarding SOPs used to generate the data;• Lack of information from the clinic regarding concomitant medications administered to individual subjects during the course of the study;• Study protocol and/or amendments not provided by the Sponsor, which may include information relevant to bioanalysis;• Discrepancies in sample management documentation provided by the sponsor or clinic and requests to make changes to correct a discrepancy without adequate supporting documentation.The survey results demonstrated that some areas of concern seem to stand out as areas for improvement. It was mentioned that the experiences of a CRO are likely to influence perceptions of risk and the frequency of exposure to areas of concern varies widely. As an industry, CROs should focus their efforts on areas of greatest frequency and risk, and use their expertise to find the right way to further discuss potential problems with the Sponsor that may be anticipated and the recommended approaches to be adopted.Feedback on the GCC & EBF recommendations on the validation of biomarkersThe topic of biomarker method validation has been a source of intense discussions as part of various GCC meetings in the past years [2,4,6]. Recommendations specific to the bioanalysis of biomarker assays were presented in several publications, including two White Papers, one prepared

The 8th GCC Closed Forum for Bioanalysis was held in Baltimore, MD, USA on 5 December 2013, immediately following the 2013 AAPS Workshop (Crystal City V): Quantitative Bioanalytical Methods Validation and Implementation--The 2013 Revised FDA Guidance. This GCC meeting was organized to discuss the contents of the draft revised FDA Guidance on bioanalytical method validation that was published in September 2013 and consolidate the feedback of the GCC members. In attendance were 63 senior-level participants, from seven countries, representing 46 bioanalytical CRO companies/sites. This event represented a unique opportunity for CRO bioanalytical experts to share their opinions and concerns regarding the draft FDA Guidance, and to build unified comments to be provided to the FDA.

BACKGROUND:To develop and validate an ultrasensitive bioanalytical assay for quantitation of fluticasone propionate in human plasma, aliquots of 0.6 ml of K(3)EDTA human plasma were treated with zinc sulfate solution and loaded onto a preconditioned SPE plate. The sample solutions were washed, eluted, dried and reconstituted. The extracted sample was injected onto a LC-MS/MS system and separated by a reverse-phase HPLC column with a 5 min gradient program, and detected by MS/MS for fluticasone propionate quantitation.RESULTS:Linearity was from 1 to 200 pg/ml. The intra- and inter-day accuracy and precision of the assay met validation acceptance criteria. Various stabilities were established and interference drug assessment was evaluated. The assay has been used for clinical studies.CONCLUSION:This ultrasensitive method has been successfully validated using LC-MS/MS for determination of fluticasone propionate in human plasma at low pg/ml level.

The decolorization kinetic of C.I. Reactive Black 5 was studied using Fenton-like oxidation. The effect of the major system parameters, such as initial pH of the solution, concentration of H2O2, concentration of Fe3+ and reaction temperature on the kinetics was determined. The results indicate that the operating parameters, such as initial pH of dye solution, initial concentration of Fe2+ and H2O2, reaction temperature, have strong influences on the degradation of C.I. Reactive Black 5 by Fenton-like oxidation process. In addition, it was also found that the decolorization of C.I. Reactive Black 5 followed the first-order kinetics well.

The Global CRO Council for Bioanalysis (GCC) was formed in September 2010. Since then, the representatives of the member companies come together periodically to openly discuss bioanalysis and the regulatory challenges unique to the outsourcing industry. The 4th GCC Closed Forum brought together experts from bioanalytical CROs to share and discuss recent issues in regulated bioanalysis, such as the impact of coadministered drugs on stability, some differences between European Medicines Agency and US FDA bioanalytical guidance documents and lessons learned following recent Untitled Letters. Recent 483s and agency findings, as well as issues on method carryover, were also part of the topics discussed.