Perspectives on gender parity in bioanalysis: an interview with Shane Needham.

Dr Shane Needham received his BS degree in chemistry from Washington State University and his PhD in chemistry from the University of Rhode Island. Dr Needham is Co-Founder and Chief Scientific Officer of Alturas Analytics, Inc. Dr Needham manages all scientific aspects of the HPLC/MS/MS bioanalytical contract laboratory at Alturas Analytics, Inc. Currently, Dr Needham's work is focused on the development and validation of assays for the determination of therapeutic agents and biomarkers from biological matrices. His laboratory leads in the area of dried blood spot analysis, microflow HPLC-MS/MS and LC-MS/MS of large molecules (Antibody Drug Conjugates, Biomarkers, New Biological Entities etc.) to support Drug Metabolism and Pharmacokinetics research.

Simultaneous extraction and analysis of multiple cystine-dense peptides by µSPE and microflow-MS/MS from plasma.

Aim: Develop a universal extraction and liquid chromatography-mass spectrometer method to simultaneously analyze cystine-dense peptide (CDP) miniproteins from rat and human plasma. The results of the analysis will be used to assist selection of therapeutic drug candidates from the vast CDP library. Methods & results: A micro-elution solid-phase extraction method was developed for the sample preparation of the CDP peptides in rat and human plasma followed by analysis by microflow liquid chromatography MS/MS. The methods developed for drug discovery were found to be accurate (±≤15.2% from nominal concentrations) and precise (≤13.4% CV), with a dynamic range of 1.00-500 ng/ml and extraction recoveries of 47.2-99.0%. Conclusion: This bioanalytical method can be utilized to screen CDP proteins and other miniproteins for drug discovery, candidate selection and further drug development.

Case Study: Absorption of Testosterone Cream via Scrotal Delivery.

Transdermal testosterone has been used for years to treat patients with low testosterone symptoms. Clinically, we have monitored patients to evaluate results of testosterone absorption via blood serum concentrations. The data on multiple time points to determine trough and peak concentrations is lacking in the literature. In this case study, we demonstrate the absorption of testosterone cream via scrotal delivery. The data suggests that after application therapeutic levels are reached with concentrations of (1204.7 ng/dL) within two hours. Additionally, consistent concentrations (1320.6 ng/dL) remain beyond six hours. To our knowledge, this is the first study to collect and measure multiple time points for testosterone via transdermal delivery. The research indicates that testosterone via transdermal delivery is an excellent method to achieve therapeutic concentrations of testosterone. Most importantly, the patient's symptoms resolved without side effects.

Clinical and Pharmaceutical Solutions through Analysis (CPSA BRASIL 2015): on the way to innovation - pharmaceutical/analytical technology, regulation and knowledge management.

Clinical and Pharmaceutical Solutions through Analysis, São Paulo, Brazil, 3-5 August 2015 The 2nd Annual Symposium on Clinical and Pharmaceutical Solutions through Analysis was held on 3-5 August 2015 at Club Transatlântico, São Paulo, Brazil. This annual meeting began in 2014 and was the first industry-led event in Brazil to focus on the specific needs of industry researchers while bringing together technology and regulators. The goal of CPSA is to provide an in-depth review of innovative technology and industry practices through open discussion of industry-related issues and needs. Education and specialized training are the foundation of all CPSA events. As the industry has evolved so has CPSA. The CPSA annual meeting thrived with high quality scientific content, open interaction from industry opinion leaders and a collegial environment.

Defining the challenges of the Modern Analytical Laboratory (CPSA USA 2014): the risks and reality of personalized healthcare.

The 17th Annual Symposium on Clinical and Pharmaceutical Solutions through Analysis (CPSA) 29 September-2 October 2014, was held at the Sheraton Bucks County Hotel, Langhorne, PA, USA. The CPSA USA 2014 brought the various analytical fields defining the challenges of the modern analytical laboratory. Ongoing discussions focused on the future application of bioanalysis and other disciplines to support investigational new drugs (INDs) and new drug application (NDA) submissions, clinical diagnostics and pathology laboratory personnel that support patient sample analysis, and the clinical researchers that provide insights into new biomarkers within the context of the modern laboratory and personalized medicine.

Conference report: Clinical and Pharmaceutical Solutions through analysis (CPSA USA 2013): connecting patients and subject numbers through analysis.

The 16th Annual Symposium on Clinical and Pharmaceutical Solutions through Analysis (CPSA) 7-10 October 2013, Sheraton Bucks County Hotel, Langhorne, PA, USA. The 2013 CPSA brought together the various US FDA regulated analytical fields affecting a 'patient' for the first time - bioanalysts supporting IND and NDAs, clinical diagnostic and pathology laboratory personnel, and clinical researchers that provide insights into new biomarkers. Although the regulatory requirements are different for each of the above disciplines, the unique analytical perspectives that affect the patient were shared - and the goal of the 2013 CPSA - 'Connecting Patients and Subject Numbers Through Analysis' was achieved.

Recent advances in the bioanalytical applications of dried matrix spotting for the analysis of drugs and their metabolites.

DBS techniques for the bioanalysis of drugs and metabolites from whole blood have been demonstrated to be a useful tool in drug development. The term dried matrix spot (DMS) has been used to indicate that the DBS technique has been applied to nonblood matrices. DMS methods often employ a color-indicating process that enhances the ability to analyze these mostly transparent fluids when spotted onto collection paper. The color-indicating dye allows the analyst to visually confirm the location of the dried sample spot. Other benefits of using a color-indicating dye include improved method accuracy and precision, because the process of adding the dye allows for the concurrent addition of the IS prior to sample addition and extraction. To date, matrices that have been analyzed using DMS include cerebrospinal fluid, synovial fluid, saliva, tears, urine and plasma.

The advantages of microflow LC-MS/MS compared with conventional HPLC-MS/MS for the analysis of methotrexate from human plasma.

BACKGROUND: In support of bioanalysis, there has always been a desire to improve detection limits and reduce scale. Microflow LC (MFLC) coupled with MS accomplishes both of these goals. RESULTS: As such, MFLC coupled with an MS system was used to generate bioanalytical validation data that met US FDA criteria. The MFLC-MS/MS data was compared with the same method with the use of conventional HPLC-MS/MS and a more than 14× S/N improvement was found with the MFLC-MS/MS method. Methotrexate was used as a model molecule to demonstrate the validation of the method from human plasma. The MFLC-MS/MS method was demonstrated to be accurate (±7%) and precise (12.9% at the LLOQ and a maximum of 11.6% at all other concentrations) across the dynamic range of the assay (1-1000 ng/ml) and compared well with the HPLC-MS/MS method. The MFLC bioanalytical validation was performed at a flow rate of 35 µl/min on a 0.5-mm inner diameter (I.D.) column, whereas, for the same linear velocities on the 2.0-mm I.D. column, the conventional HPLC bioanalytical validation was performed at 700 µl/min. Since the flow rate of the MFLC system is 20-times less than the HPLC system, the consumable solvent and disposal cost to perform the MFLC validation was significantly less. CONCLUSION: MFLC-MS/MS can be used to perform bioanalytical method validations with increased MS signal, reduced source contamination and reduced solvent consumption.

Transforming drug discovery and development through innovation: past, present and future.

This annual meeting began in 1998 and was the first industry-led event to focus on the specific needs of industry researchers. The goal of Clinical and Pharmaceutical Solutions Through Analysis (CPSA) is to provide an in-depth review of innovative technology and industry practices through open discussion of industry-related issues and needs. Education and specialized training are the foundation of all CPSA events. As the industry has evolved so has CPSA. Thus, the year the name was changed from Chemical and Pharmaceutical Structural Analysis to CPSA was to reflect the growing focus on clinical applications and the emergence of personalized medicine. Most importantly, the CPSA annual meeting has retained the same high-quality scientific content, open interaction from industry opinion leaders and a collegial environment.

Recommendations on bioanalytical method stability implications of co-administered and co-formulated drugs by Global CRO Council for Bioanalysis (GCC).

An open letter written by the Global CRO Council for Bioanalysis (GCC) describing the GCC survey results on stability data from co-administered and co-formulated drugs was sent to multiple regulatory authorities on 14 December 2011. This letter and further discussions at different GCC meetings led to subsequent recommendations on this topic of widespread interest within the bioanalytical community over the past 2 years.

4th Global CRO Council for Bioanalysis: coadministered drugs stability, EMA/US FDA guidelines, 483s and carryover.

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.

Science and technology coming together to make a difference.

This annual meeting began in 1998 and was the first industry-led event to focus on the specific needs of industry researchers. The goal of Chemical and Pharmaceutical Structure Analysis is to provide an in-depth review of innovative technology and industry practices through open discussion of industry-related issues and needs. Education and specialized training are the foundation of all Chemical and Pharmaceutical Structure Analysis events - where technology and solutions meet.

Use of conventional bioanalytical devices to automate DBS extractions in liquid-handling dispensing tips.

BACKGROUND: Conventional liquid-handling devices were employed, along with an improved punching device, to semi-automate dried blood spot (DBS) extraction of alprazolam, α-hydroxyalprazolam and midazolam from human whole blood. Liquid-handling devices were used to add internal standard to the DBS cards and to extract the analytes from the DBS, in order to be analyzed by HPLC-MS/MS. RESULTS: The technique was shown to be accurate (±12.0%) and precise (10.3%) across the dynamic range of the assay. CONCLUSION: The semi-automated extraction reduced sample preparation time by more than 50% when compared with more conventional DBS manual extraction methods.

Recommendations on: internal standard criteria, stability, incurred sample reanalysis and recent 483s by the Global CRO Council for Bioanalysis.

"The Global CRO Council (GCC) for Bioanalysis was formed in an effort to bring together many CRO leaders to openly discuss bioanalysis and the regulatory challenges unique to the outsourcing industry"

Validation of HPLC-MS/MS methods for analysis of loxapine, amoxapine, 7-OH-loxapine, 8-OH-loxapine and loxapine N-oxide in human plasma.

BACKGROUND: Two ESI-LC-MS/MS methods were validated for the quantitative analysis of loxapine, amoxapine, 7-OH-loxapine, 8-OH-loxapine and loxapine N-oxide in human K(2)EDTA plasma. Cation-exchange solid-phase extraction (SPE) was used to extract loxapine, amoxapine and the two hydroxylated metabolites, and organic precipitation was used to quantify loxapine N-oxide. RESULTS: Both methods were shown to be accurate (±13%), intra-assay precision was less than 15%, and inter-assay precision was less than 10% in all instances across the entire dynamic range of the assays (0.0500-50.0 ng/ml for the SPE method and 0.100-25.0 ng/ml for the precipitation method). CONCLUSION: The validated methods for loxapine, amoxapine, 7-OH-loxapine, 8-OH-loxapine and loxapine N-oxide have been used to successfully support clinical trials.