Sotorasib is a small molecule KRASG12C inhibitor approved for the treatment of KRASG12C mutated locally advanced or metastatic non-small cell lung cancer in adult patients. The effect of hepatic impairment on the pharmacokinetics (PK) of sotorasib and major metabolites M10, M18, and M24, safety, and tolerability after a single oral dose of 960 mg was assessed in a phase 1, parallel-arm, multi-center (US), open-label study. Sotorasib AUCinf ratio for subjects with moderate (n = 7) or severe (n = 4) hepatic impairment relative to normal hepatic function (n = 7) was 0.746 (90% CI: 0.431-1.29) and 1.04 (0.545-1.97), respectively. Cmax ratio for moderate and sever hepatic impairment was 0.955 (0.512-1.78) and 1.43 (0.688-2.96), respectively. Mean t1/2 values for sotorasib were similar in subjects with normal hepatic function and subjects with moderate or severe hepatic impairment. Cmax and AUCinf of M10 and M24 increased, while M18 decreased with increasing severity of hepatic impairment. Treatment-emergent adverse events were mild in severity and no serious adverse events were reported. Overall, moderate or severe hepatic impairment did not considerably affect the exposure of sotorasib, M10, M18, and M24. This data supports that adjustments to sotorasib dosing are not indicated for moderate or severe hepatic impairment.

BioanalysisVol. 15, No. 3 CommentaryPerspectives on a flexible strategy for the management of nonregulated bioanalysisBernd A Bruenner, Roger V Pham, Wale Rufai & Christopher A JamesBernd A Bruenner *Author for correspondence: E-mail Address: berndb@amgen.comhttps://orcid.org/0000-0002-3785-6035Bioanalytical Sciences, Amgen, One Amgen Center Dr., Thousand Oaks, CA 91320-1799, USASearch for more papers by this author, Roger V PhamBioanalytical Sciences, Amgen, One Amgen Center Dr., Thousand Oaks, CA 91320-1799, USASearch for more papers by this author, Wale RufaiBioanalytical Sciences, Amgen, One Amgen Center Dr., Thousand Oaks, CA 91320-1799, USASearch for more papers by this author & Christopher A James https://orcid.org/0000-0001-8749-0011Bioanalytical Sciences, Amgen, One Amgen Center Dr., Thousand Oaks, CA 91320-1799, USASearch for more papers by this authorPublished Online:9 Feb 2023https://doi.org/10.4155/bio-2022-0094AboutSectionsView ArticleView Full TextPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInReddit View articleKeywords: bioanalysisnonregulatedoutsourcingqualityReferences1. Xu RN, Fan L, Rieser MJ, El-Shourbagy TA. Recent advances in high-throughput quantitative bioanalysis by LC–MS/MS. J. Pharm. Biomed. Anal. 44(2), 342–355 (2007).Crossref, Medline, CAS, Google Scholar2. Varma D, Jansen SA, Ganti S. Chromatography with higher pressure, smaller particles and higher temperature: a bioanalytical perspective. Bioanalysis 2(12), 2019–2034 (2010).Link, CAS, Google Scholar3. Smith NF, Raynaud FI, Workman P. The application of cassette dosing for pharmacokinetic screening in small-molecule cancer drug discovery. Mol. Cancer Ther. 6(2), 428–440 (2007).Crossref, Medline, CAS, Google Scholar4. Ho S. Best practices for discovery bioanalysis: balancing data quality and productivity. Bioanalysis 6(20), 2705–2708 (2014).Link, CAS, Google Scholar5. Timmerman P, White S, Mcdougall S et al. Tiered approach into practice: scientific validation for chromatography-based assays in early development – a recommendation from the European Bioanalysis Forum. Bioanalysis 7(18), 2387–2398 (2015).Link, CAS, Google Scholar6. Sydor J, Kim G. Are we ready for the evolution of electronic laboratory notebooks in regulated bioanalysis? Bioanalysis 9(16), 1203–1205 (2017).Link, CAS, Google Scholar7. Verhaeghe T. Bioanalytical outsourcing strategy at Janssen Research and Development. Bioanalysis 6(10), 1321–1327 (2014).Link, CAS, Google Scholar8. Soto M, Pham R, Almon V et al. Evaluation of matrix microsampling methods for therapeutic drug candidate quantification in discovery-stage rodent pharmacokinetic studies. Bioanalysis 6(16), 2135–2146 (2014).Link, CAS, Google Scholar9. Uyeda C, Pham R, Fide S et al. Application of automated dried blood spot sampling and LC–MS/MS for pharmacokinetic studies of AMG 517 in rats. Bioanalysis 3(20), 2349–2356 (2011).Link, CAS, Google Scholar10. Velghe S, Delahaye L, Stove CP. Is the hematocrit still an issue in quantitative dried blood spot analysis? J. Pharm. Biomed. Anal. 163, 188–196 (2019).Crossref, Medline, CAS, Google Scholar11. Lavelle A, Micheli J. COVID-19’s impact on bioanalytical labs. Bioanalysis 13(15), 1169–1171 (2021).Link, CAS, Google ScholarFiguresReferencesRelatedDetailsCited ByIntroducing the 2023 Bioanalysis special-themed issue on laboratory structure and management for nonregulated bioanalysisQin C Ji & Gary J Jenkins30 March 2023 | Bioanalysis, Vol. 15, No. 3 Vol. 15, No. 3 STAY CONNECTED Metrics Downloaded 110 times History Received 12 May 2022 Accepted 10 October 2022 Published online 9 February 2023 Published in print February 2023 Information© 2023 Newlands PressKeywordsbioanalysisnonregulatedoutsourcingqualityFinancial & competing interests disclosureThe authors are employees of and hold stock in Amgen, Inc. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.No writing assistance was utilized in the production of this manuscript.PDF download

Background: Sotorasib (AMG 510) is a first-in-class KRAS(G12C) inhibitor that received accelerated US FDA approval in 2021 for the treatment of patients with KRAS(G12C)-mutated locally advanced or metastatic non-small-cell lung cancer. Method: An LC-MS/MS method was developed and validated for the determination of sotorasib in human plasma to support clinical development studies. Samples were prepared using protein precipitation and analyzed by LC-MS/MS using gradient elution with a calibration standard curve range of 10.0-10,000 ng/ml. Stable isotope labeled [C-13, D-3]-sotorasib was used as an internal standard. Results and conclusion: The method fully met FDA guidelines for all validation parameters, including precision, accuracy, selectivity, matrix effect, recovery and stability and has been extensively used to support multiple clinical studies.

The 15th edition of the Workshop on Recent Issues in Bioanalysis (15th WRIB) was held on 27 September to 1 October 2021. Even with a last-minute move from in-person to virtual, an overwhelmingly high number of nearly 900 professionals representing pharma and biotech companies, contract research organizations (CROs), and multiple regulatory agencies still eagerly convened to actively discuss the most current topics of interest in bioanalysis. The 15th WRIB included 3 Main Workshops and 7 Specialized Workshops that together spanned 1 week in order to allow exhaustive and thorough coverage of all major issues in bioanalysis, biomarkers, immunogenicity, gene therapy, cell therapy and vaccines. Moreover, in-depth workshops on biomarker assay development and validation (BAV) (focused on clarifying the confusion created by the increased use of the term "Context of Use - COU"); mass spectrometry of proteins (therapeutic, biomarker and transgene); state-of-the-art cytometry innovation and validation; and, critical reagent and positive control generation were the special features of the 15th edition. This 2021 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2021 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 1A) covers the recommendations on Endogenous Compounds, Small Molecules, Complex Methods, Regulated Mass Spec of Large Molecules, Small Molecule, PoC. Part 1B covers the Regulatory Agencies' Inputs on Bioanalysis, Biomarkers, Immunogenicity, Gene & Cell Therapy and Vaccine. Part 2 (ISR for Biomarkers, Liquid Biopsies, Spectral Cytometry, Inhalation/Oral & Multispecific Biotherapeutics, Accuracy/LLOQ for Flow Cytometry) and Part 3 (TAb/NAb, Viral Vector CDx, Shedding Assays; CRISPR/Cas9 & CAR-T Immunogenicity; PCR & Vaccine Assay Performance; ADA Assay Comparabil ity & Cut Point Appropriateness) are published in volume 14 of Bioanalysis, issues 10 and 11 (2022), respectively.

Advances in the technologies to enable patient-centric sampling (PCS) have the potential to improve blood sample collection by enabling clinical trial participants to collect samples via self-collection or with the help of a caregiver in their home. Typically, blood samples to assess pharmacokinetics and pharmacodynamics of a drug during clinical development are collected at a clinical site via venous blood draw. In this position paper by the International Consortium for Innovation and Quality in Pharmaceutical Development (IQ), the potential value PCS can bring to patients, to the clinical datasets generated, and to clinical trial sponsors is discussed, along with considerations for program decision making, bioanalytical feasibility, operations, and regulatory implications. With an understanding of the value of PCS and considerations when implementing during clinical drug development, we can bring the promise of PCS closer to reality and enable decentralized clinical trials.

BioanalysisVol. 13, No. 5 EditorialBioanalysis and the oncology revolutionEnaksha R Wickremsinhe & Christopher A JamesEnaksha R Wickremsinhe*Author for correspondence: E-mail Address: enaksha@lilly.comEli Lilly & Company, Indianapolis, IN 46285, USASearch for more papers by this author & Christopher A JamesAmgen Research, Thousand Oaks, CA 91320, USASearch for more papers by this authorPublished Online:4 Mar 2021https://doi.org/10.4155/bio-2021-0014AboutSectionsView ArticleView Full TextPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInReddit View articleKeywords: bioanalyticalco-med interferenceco-med stabilitynovel drug modalitiesoncology studiespolypharmacyselectivitystability testingReferences1. Martin-Liberal J, Hierro C, Ochoa De Olza M, Rodon J. Immuno-oncology: the third paradigm in early drug development. Target Oncol. 12(2), 125–138 (2017).Crossref, Medline, Google Scholar2. 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A randomized, 3-arm, neoadjuvant, Phase II study comparing docetaxel + carboplatin + trastuzumab + pertuzumab (TCbHP), TCbHP followed by trastuzumab emtansine and pertuzumab (T-DM1 + P), and T-DM1 + P in HER2-positive primary breast cancer. Breast Cancer Res. Treat. 180(1), 135–146 (2020).Crossref, Medline, CAS, Google Scholar8. Wan X, Luo X, Tan C, Zeng X, Zhang Y, Peng L. First-line atezolizumab in addition to bevacizumab plus chemotherapy for metastatic, nonsquamous non-small-cell lung cancer: A United States-based cost–effectiveness analysis. Cancer 125(20), 3526–3534 (2019).Crossref, Medline, CAS, Google Scholar9. Mateos M-V, Cavo M, Bladé J et al. Daratumumab plus bortezomib, melphalan, and prednisone versus bortezomib, melphalan, and prednisone in patients with transplant-ineligible newly diagnosed multiple myeloma: overall survival in Alcyone. Blood 134(Suppl. 1), 859 (2019).Crossref, Google Scholar10. O'malley DM, Richardson DL, Vergote IB et al. Mirvetuximab soravtansine (MIRV), a folate receptor alpha (FRa)-targeting antibody-drug conjugate (ADC), in combination with carboplatin (CARBO) and bevacizumab (BEV): final results from a study in patients (pts) with recurrent platinum sensitive ovarian cancer. Ann. Oncol. 31, S626–S627 (2020).Medline, Google Scholar11. Paller CJ, Huang EP, Luechtefeld T et al. Factors affecting combination trial success (FACTS): investigator survey results on Early-Phase Combination Trials. Front Med. (Lausanne) 6, 122 (2019).Crossref, Medline, Google Scholar12. Bergholz JS, Wang Q, Kabraji S, Zhao JJ. Integrating immunotherapy and targeted therapy in cancer treatment: mechanistic insights and clinical implications. Clin. Cancer Res. 26(21), 5557–5566 (2020).Crossref, Medline, CAS, Google Scholar13. Sicklick JK, Kato S, Okamura R et al. Molecular profiling of cancer patients enables personalized combination therapy: the I-PREDICT study. Nat. 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In: Antibody-Drug Conjugates: Methods and Protocols. Tumey LN (Ed.). Springer, NY, USA (2020).Crossref, Google Scholar19. Liu J, Li J, Tran C et al. Oligonucleotide quantification and metabolite profiling by high-resolution and accurate mass spectrometry. Bioanalysis 11(21), 1967–1980 (2019).Link, CAS, Google Scholar20. Spengler M, Adler M, Niemeyer CM. Highly sensitive ligand-binding assays in pre-clinical and clinical applications: immuno-PCR and other emerging techniques. Analyst 140(18), 6175–6194 (2015).Crossref, Medline, CAS, Google Scholar21. De Zwart M, Lausecker B, Globig S et al. Co-medication and interference testing in bioanalysis: a European Bioanalysis Forum recommendation. Bioanalysis 8(19), 2065–2070 (2016).Link, Google Scholar22. US FDA. Bioanalytical Method Validation Guidance for Industry (2018). https://www.fda.gov/downloads/drugs/guidances/ucm070107.PdfGoogle Scholar23. European Medicines Agency. 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Bioanalysis 11(20), 1819–1821 (2019).Link, CAS, Google ScholarFiguresReferencesRelatedDetailsCited ByNanocatalysis meets microfluidics: A powerful platform for sensitive bioanalysisTrAC Trends in Analytical Chemistry, Vol. 1582021 White Paper on Recent Issues in Bioanalysis: Mass Spec of Proteins, Extracellular Vesicles, CRISPR, Chiral Assays, Oligos; Nanomedicines Bioanalysis; ICH M10 Section 7.1; Non-Liquid & Rare Matrices; Regulatory Inputs (Part 1A – Recommendations on Endogenous Compounds, Small Molecules, Complex Methods, Regulated Mass Spec of Large Molecules, Small Molecule, PoC & Part 1B - Regulatory Agencies' Inputs on Bioanalysis, Biomarkers, Immunogenicity, Gene & Cell Therapy and Vaccine)Surinder Kaur, Stephen C Alley, Matt Szapacs, Amanda Wilson, Eugene Ciccimaro, Dian Su, Neil Henderson, Linzhi Chen, Fabio Garofolo, Shawna Hengel, Wenying Jian, John F Kellie, Anita Lee, John Mehl, Joe Palandra, Haibo Qiu, Natasha Savoie, Diaa Shakleya, Ludovicus Staelens, Hiroshi Sugimoto, Giane Sumner, Jan Welink, Robert Wheller, Y-J Xue, Jianing Zeng, Jinhui Zhang, Huiyu Zhou, Jian Wang, Scott Summerfield, Olga Kavetska, Lieve Dillen, Ragu Ramanathan, Mike Baratta, Arindam Dasgupta, Anna Edmison, Luca Ferrari, Sally Fischer, Daniela Fraier, Sam Haidar, Kathrin Heermeier, Christopher James, Allena Ji, Lina Luo, Gustavo Mendes Lima Santos, Noah Post, Anton I Rosenbaum, Sune Sporring, Sekhar Surapaneni, Stephen Vinter, Katty Wan, Eric Woolf, Olga Kavetska, Seongeun (Julia) Cho, Elham Kossary, Sandra Prior, Mohsen Rajabi Abhari, Catherine Soo, Yow-Ming Wang, Abbas Bandukwala, Elana Cherry, Isabelle Cludts, Soma Ghosh, Shirley Hopper, Akiko Ishii-Watabe, Susan Kirshner, Kevin Maher, Kimberly Maxfield, Joao Pedras-Vasconcelos, Yoshiro Saito, Dean Smith, Therese Solstad, Daniela Verthelyi, Meenu Wadhwa, Leslie Wagner, Günter Waxenecker, Haoheng Yan & Lucia Zhang17 May 2022 | Bioanalysis, Vol. 14, No. 9Welcome to volume 14 of BioanalysisSankeetha Nadarajah29 November 2021 | Bioanalysis, Vol. 14, No. 1Bioanalytical Challenges due to Prior Checkpoint Inhibitor Exposure: Interference and Mitigation in Drug Concentration and Immunogenicity Assays4 October 2021 | The AAPS Journal, Vol. 23, No. 6Quantification of abemaciclib and metabolites: evolution of bioanalytical methods supporting a novel oncolytic agentEnaksha R Wickremsinhe & Lisa B Lee19 April 2021 | Bioanalysis, Vol. 13, No. 9 Vol. 13, No. 5 Follow us on social media for the latest updates Metrics Downloaded 227 times History Received 17 January 2021 Accepted 17 February 2021 Published online 4 March 2021 Published in print March 2021 Information© 2021 Newlands PressKeywordsbioanalyticalco-med interferenceco-med stabilitynovel drug modalitiesoncology studiespolypharmacyselectivitystability testingAcknowledgmentsThe authors would like to thank A Wolf, P Conliffe, B Ackermann and K Cox for helpful suggestions.Financial & competing interests disclosureThe authors of this article are current employees of their affiliated organizations. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.No writing assistance was utilized in the production of this manuscript.PDF download

The use of quality control (QC) samples in bioanalysis is well established and consistent with regulatory guidance. However, a systematic evaluation of whether QC samples serve the intended purpose of improving data quality has not been undertaken. The Translational and ADME Sciences Leadership Group (TALG) of the International Consortium for Innovation and Quality in Pharmaceutical Development (IQ) conducted an evaluation to assess whether closer agreement is observed when comparing pharmacokinetic data from two passed runs, than when comparing data from failed and passed (retest) runs. Analysis of data collected across organizations, molecular types and analytical platforms, revealed that bioanalytical methods are very reproducible; and that QC samples improve the overall quality of pharmacokinetic concentration data and justifies their continued use.

The 2019 13th Workshop on Recent Issues in Bioanalysis (WRIB) took place in New Orleans, LA on 1-5 April 2019 with an attendance of over 1000 representatives from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. WRIB was once again a 5-day, week-long event - a full immersion week of bioanalysis, biomarkers, immunogenicity and gene therapy. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small- and large-molecule bioanalysis involving LCMS, hybrid LBA/LCMS, LBA cell-based/flow cytometry assays and qPCR approaches. This 2019 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2019 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 2) covers the recommendations on the 2018 FDA BMV guidance, 2019 ICH M10 BMV draft guideline and regulatory agencies' input on bioanalysis, biomarkers, immunogenicity and gene therapy. Part 1 (Innovation in small molecules and oligonucleotides and mass spectrometry method development strategies for large molecules bioanalysis) and Part 3 (New insights in biomarker assay validation, current and effective strategies for critical reagent management, flow cytometry validation in drug discovery and development and CLSI H62, interpretation of the 2019 FDA immunogenicity guidance and gene therapy bioanalytical challenges) are published in volume 10 of Bioanalysis, issues 22 and 24 (2019), respectively.

The 2018 12th Workshop on Recent Issues in Bioanalysis (12th WRIB) took place in Philadelphia, PA, USA on April 9-13, 2018 with an attendance of over 900 representatives from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. WRIB was once again a 5-day full immersion in bioanalysis, biomarkers and immunogenicity. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small- and large-molecule bioanalysis involving LC-MS, hybrid ligand binding assay (LBA)/LC-MS and LBA/cell-based assays approaches. This 2018 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2018 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 1) covers the recommendations for LC-MS for small molecules, peptides, oligonucleotides and small molecule biomarkers. Part 2 (hybrid LBA/LC-MS for biotherapeutics and regulatory agencies' inputs) and Part 3 (large molecule bioanalysis, biomarkers and immunogenicity using LBA and cell-based assays) are published in volume 10 of Bioanalysis, issues 23 and 24 (2018), respectively.

Quantitative analysis of therapeutic monoclonal antibodies (mAbs) in biological matrices has become increasingly important with the growing number of novel therapeutic antibody-based therapeutics. Liquid chromatography-mass spectrometry (LC-MS/ MS) is emerging as a promising alternative and complementary assay platform for quantitative analysis of therapeutic antibodies. This chapter highlights some recent method development strategies and provides case studies to represent utility of mAb selective enrichment approach in support of preclinical therapeutic mAb discovery research. Surrogate peptides serve as the accurate quantitative. The strategies for sample preparation of antibody LC-MS/MS bioanalysis are divided into two main categories: immunoaffinity-based sample preparation and non-immunoaffinity-based sample preparation. In general, sample cleanup requirements are more challenging for antibody quantitation than for small-molecule LC-MS/MS bioanalysis. The use of internal standards is a near universal practice in quantitative LC-MS/MS bioanalysis to ensure assay accuracy, precision, and reproducibility.

A LC-MS/MS method has been developed and validated for the determination of glycine in human cerebrospinal fluid (CSF). The validated method used artificial cerebrospinal fluid as a surrogate matrix for calibration standards. The calibration curve range for the assay was 100-10,000 ng/mL and C-13(2), N-15-glycine was used as an internal standard (IS). Pre-validation experiments were performed to demonstrate parallelism with surrogate matrix and standard addition methods. The mean endogenous glycine concentration in a pooled human CSF determined on three days by using artificial CSF as a surrogate matrix and the method of standard addition was found to be 748 +/- 30.6 and 768 +/- 18.1 ng/mL, respectively. A percentage difference of -2.6% indicated that artificial CSF could be used as a surrogate calibration matrix for the determination of glycine in human CSF. Quality control (QC) samples, except the lower limit of quantitation (LLOQ) QC and low QC samples, were prepared by spiking glycine into aliquots of pooled human CSF sample. The low QC sample was prepared from a separate pooled human CSF sample containing low endogenous glycine concentrations, while the LLOQ QC sample was prepared in artificial CSF. Standard addition was used extensively to evaluate matrix effects during validation. The validated method was used to determine the endogenous glycine concentrations in human CSF samples. Incurred sample reanalysis demonstrated reproducibility of the method. (C) 2016 Elsevier B.V. All rights reserved.

The 2016 10th Workshop on Recent Issues in Bioanalysis (10th WRIB) took place in Orlando, Florida with participation of close to 700 professionals from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations, and regulatory agencies worldwide. WRIB was once again a 5-day, weeklong event - A Full Immersion Week of Bioanalysis including Biomarkers and Immunogenicity. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small and large molecule analysis involving LCMS, hybrid LBA/LCMS, and LBA approaches, with the focus on biomarkers and immunogenicity. This 2016 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. This white paper is published in 3 parts due to length. This part (Part 1) discusses the recommendations for small molecules, peptides and small molecule biomarkers by LCMS. Part 2 (Hybrid LBA/LCMS and regulatory inputs from major global health authorities) and Part 3 (large molecule bioanalysis using LBA, biomarkers and immunogenicity) will be published in the Bioanalysis journal, issue 23.

This paper was developed with the support of the International Consortium for Innovation and Quality in Pharmaceutical Development (IQ). IQ is a not-for-profit organization of pharmaceutical and biotechnology companies with a mission of advancing science-based and scientifically driven standards and regulations for pharmaceutical and biotechnology products worldwide. Within the IQ, various working groups (WG) have been formed, where the microsampling WG is committed to providing a scientific forum for the advancement of both wet and dry microsampling techniques within the pharmaceutical industry. This first output from the microsampling WG is to summarize and reflect on the current knowledge and opinions on DBS sampling, to stimulate discussion, and to encourage future creative applications of DBS sampling. Dried blood spot (DBS) sampling has established itself as an innovative sampling technique where wet blood is spotted onto absorbent paper or other paper materials and allowed to dry (1–4). DBS offers several potential benefits inherent to the technique, namely a low blood volume, simplified blood sample collection (5), and convenient sample storage and transfer. In certain applications, DBS sampling has been shown to stabilize certain analytes or metabolites without the addition of chemical modifiers (6–9). DBS has been routinely applied for decades in neonatal screening for phenylketonuria and other congenital metabolic disorders (10). The utility of DBS sampling has also been demonstrated for therapeutic drug monitoring (11) and for epidemiological studies (e.g., HIV and HBV detection/monitoring) (12) due to the practical advantages along with simplified sample collection and handling procedures. Finally, DBS can also be used for quantitative biomarker (PD) assessment from blood, where appropriate. However, the technique is relatively new to the pharmaceutical industry and to government regulators overseeing new drug applications. Nevertheless, over the past 5 to 7 years, the technique has been extensively evaluated for quantifying drug exposure in nonclinical and/or clinical studies in various stages of drug discovery and development. The ease to collect, transfer, store, and process small volumes of blood samples has generated considerable interest in providing utility in volume-limited situations (e.g., small rodent, human pediatric studies) for toxicokinetic (TK), pharmacokinetic (PK), or pharmacodynamic (PD) sampling. Discovery and nonclinical studies Rodent animal models are typically employed in these studies. The reduced blood volumes required for DBS can enable serial bleeding and, consequently, elimination of satellite animal groups and reduction of compound use. The ability to eliminate the satellite animal groups enables the assessment of exposure and toxic effects within the same animal. Studies involving expensive animal models (i.e., transgenic mice, knock-out mice, humanized mice, etc.) further highlight a persuasive scientific and economic case for DBS sampling since a complete pharmacokinetic profile can be obtained from a single study animal without the need for extra rodents merely for generating exposure data. These are perfectly in line with the principles of the 3Rs: reduction, refinement, and replacement of humane animal research (13–15). With greater emphasis from the regulatory authorities to study new drugs for infants, neonates, and pediatric populations, the requirement to conduct associated nonclinical juvenile rodent toxicity studies serves as an ideal scenario where the advantage of low blood volume in DBS sampling is undeniable. Although the advantages of DBS heavily favor rodent studies, it can also be used to refine non-rodent studies.

BioanalysisVol. 7, No. 8 EditorialHow can chromatographic baseline integration parameters be objectively established in the bioanalytical laboratory?Christopher A James, Bernd A Bruenner & Philip WongChristopher A JamesAuthor for correspondence: E-mail Address: cajames@amgen.comAmgen, Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USASearch for more papers by this author, Bernd A BruennerAmgen, Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USASearch for more papers by this author & Philip WongAmgen, Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USASearch for more papers by this authorPublished Online:12 May 2015https://doi.org/10.4155/bio.15.33AboutSectionsView ArticleView Full TextPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInRedditEmail View articleKeywords: baselinesbioanalysischromatographyintegrationLC–MS/MSreintegrationvalidationReferences1 Garofolo F, Rocci ML, Dumont I et al. White paper on recent issues in bioanalysis and regulatory findings from audits and inspections. Bioanalysis 3(18), 2081–2096 (2011).Link, CAS, Google Scholar2 Hill HM, Smeraglia J, Brodie RR, Smith GT. Where do you draw the line? Points to consider when implementing the FDA method Guidance on quantifying chromatographic peaks. Chromatographia 55(1 Suppl. 1), S79–S81 (2002).Crossref, CAS, Google Scholar3 Smith G, James CA, Scitt R, Woolf E. Chromatographic baselines. Bioanalysis 6(9), 1167–1170 (2014).Link, CAS, Google Scholar4 Hill HM, Bakes D, Love I. Manual chromatographic baseline integration: is it needed, if so when should it be used? Bioanalysis 6(9), 1171–1174 (2014).Link, CAS, Google Scholar5 European Medicines Agency. Guideline on bioanalytical method validation 2011. www.ema.europa.eu/docs/en_GB/document_library/Scientific_ guideline/2 011/08/WC500109686.pdf.Google Scholar6 US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM), May 2001. Guidance for Industry Bioanalytical Method Validation www.fda.gov/downloads/Drugs/Guidances/ucm070107.pdf.Google Scholar7 Woolf EJ, McDougall S, Fast DM et al. Small molecule specific run acceptance, specific assay operation, and chromatographic run quality assessment: recommendation for best practices and harmonization from the global bioanalysis consortium harmonization teams. AAPS J. 16(5), 885–893 (2014).Crossref, Medline, CAS, Google ScholarFiguresReferencesRelatedDetailsCited ByQuantitative glycomics using liquid phase separations coupled to mass spectrometry1 January 2017 | The Analyst, Vol. 142, No. 5 Vol. 7, No. 8 STAY CONNECTED Metrics Downloaded 212 times History Published online 12 May 2015 Published in print May 2015 Information© Future Science LtdKeywordsbaselinesbioanalysischromatographyintegrationLC–MS/MSreintegrationvalidationFinancial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.PDF download

Analysis of pharmaceutical compounds in cerebrospinal fluid (CSF) may present challenges due to the combination of the low protein content in this matrix and relatively low drug concentrations, often corresponding to free drug concentrations in plasma, typically found in CSF. A 30% loss of AMG 579 was observed during preparation of quality control samples and further investigation determined that this loss was likely due to binding to collection tubes. This observation also highlighted the possibility of additional losses of AMG 579 that could occur during collection of clinical samples, such as binding to catheters used in the collection of CSF.Loss of AMG 579 in QC samples was reduced from 30% to 5% when the volume of CSF stored in 1.5 mL vials was increased from 0.06 mL to 1 mL. Modest but unavoidable losses of about 20% of AMG 579 were also found following perfusion through both silicone and polypropylene (Pharmed (R) BPT) collection catheters. Silicone tubing was used for CSF collection based on clinical site preference. An LC-MS/MS method was validated to quantify AMG 579 in human CSF to support clinical testing. The original range of the assay was 1-1000 ng/mL but the LLOQ was subsequently lowered to 0.1 ng/mL to better meet project requirements. Interday bias (% RE) and precision (% CV) were -4.2% and 12.3% at the LLOQ and less than +/- 0.9% and 8.3% for higher concentrations, respectively. The compound was stable in human CSF for at least 5h at room temperature, 55 days at -70 degrees C (-60 to -80 degrees C range), and through three freeze-thaw cycles.Careful selection of assay conditions and materials minimized losses of the compound during sample collection and storage. While these losses could not be entirely eliminated, practical sample collection and storage conditions were established to allow for analysis of AMG 579 in human clinical trials. (C) 2015 Elsevier B.V. All rights reserved.

Future Science Book SeriesAdvanced LC-MS Applications in Bioanalysis Complex sample typesBernd A Bruenner, Mary C Wells & Christopher A JamesBernd A Bruenner, Mary C Wells & Christopher A JamesPublished Online:28 Oct 2015https://doi.org/10.4155/fseb2013.13.131AboutSectionsView ArticleView Full TextPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinkedInReddit View chapterReferences1. Bateman KP , Castonguay G , Xu L et al. Reduction of animal usage by serial bleeding of mice for pharmacokinetic studies: application of robotic sample preparation and fast liquid chromatography-mass spectrometry . J. Chromatogr. B Biomed. Sci. Appl. 754 ( 1 ), 245 – 251 ( 2001 ). Crossref, Medline, CAS, Google Scholar2. Shen DD , Artru AA , Adkison KK . Principles and applicability of CSF sampling for the assessment of CNS drug delivery and pharmacodynamics . Adv. Drug Deliv. Rev. 56 ( 12 ), 1825 – 1857 ( 2004 ). Crossref, Medline, CAS, Google Scholar3. 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Crossref, Medline, CAS, Google ScholarFiguresReferencesRelatedDetails Advanced LC-MS Applications in BioanalysisMetrics Downloaded 15 times History Published online 28 October 2015 Published in print October 2015 Information© Future Science Ltd© Future Science LtdPDF download

As part of this themed issue, Bioanalysis invited a selection of researchers to express their views on method transfer in the bioanalytical field. The topics discussed include the main challenges in method transfer, the importance of communication and approaches for assessing the acceptability of transfers. Their responses provide a valuable insight into the considerations that need to be taken into account for successful method transfer.

Background: AMG 517 or 1-aminobenzotriazole were quantified by LC-MS/MS from low blood/plasma volumes for rat pharmacokinetic (PK) characterization in order to qualify manual/automated dried blood spot (DBS) sampling and plasma separation capillary sampling. In addition, mouse serial automated blood sampling was compared with standard composite sampling. Materials & methods: AMG 517 or 1-aminobenzotriazole was administered to rats or mice and multiple microsampling techniques were used to obtain blood or plasma. Results: PK parameters derived from DBS and whole blood-obtained drug concentrations were within 7% for manual DBS and 20% for automated DBS. Plasma PK parameters derived from capillary or standard plasma-obtained drug concentrations differed by 6%. Plasma PK parameters obtained from serial automated blood sampling or manual composite sampling were within 20%. Conclusion: Collectively, these results suggest that the microsampling applications that were investigated are attractive approaches for quantifying drug candidates in low matrix volumes that can be successfully employed within discovery-stage rodent PK studies.

Bioanalysis invited a selection of leading researchers to express their views on chromatographic baseline assignment in the bioanalytical laboratory. The topics discussed include the challenges presented with ensuring automated baseline assignment is correct, when reintegration is necessary, regulation and consistency in terminology. Their enlightening responses provide a valuable insight into developing an industry consensus towards reintegration. An accompanying commentary article in this issue, authored by Howard Hill and colleagues (Huntingdon Life Sciences), provides background to this much debated topic.