Comprehensive Bioanalytical Support for Research and Innovation
Mission
The Bioanalytical Core, located within the Portland Laboratory for Biotechnology and Health Sciences (PLBHS), provides a comprehensive approach to absorption, distribution, metabolism and excretion (ADME) models, as well as pharmacokinetic and bioanalytical assay development to support members of the 91AV research community, as well as external clients. We use the highest level of expertise, training, and instrumentation (LC-MS/MS) to enable moderate throughput pharmacology, endocrine and drug metabolism assays with applications in drug discovery, translational biology, toxicology and proof-of-concept studies. Staff have significant pharmacology and Drug Metabolism and Pharmacokinetics (DMPK) experience from both academic and corporate (pharma) research environments, enabling quick method development and analysis for diverse types of projects.
Bioanalytical Core Leadership
Director, Bioanalytical Core
Professor of Pharmacology
Vice President for Research
Bioanalytical Research Specialist
Areas of Expertise
Drug Discovery
ADME Models
- Aqueous solubility
- LogD
- Microsomal stability
- Small molecule determinations following in vivo dosing
- Identification of small molecule metabolites
- Plasma protein binding
Validated Biomarker Quantification
- Bioanalytical quantification of iron species, lipids, and corticosteroids in tissues and biofluids
- Custom assay development for the quantification of drugs and biomarkers in diverse biological tissues such as plasma, serum, brain, spinal cord, and bone
Pharmacokinetics
- Single-dose pharmacokinetic assessment with non-compartmental analysis
Translational Pharmacology
- Expertise in assay development for the quantification of drugs and biomarkers in diverse biological tissues such as plasma, serum, brain, spinal cord, liver, and bone
- Tissue niche exposure of many drug classes, including antipsychotics, opioids, and beta blockers in pre-clinical models
Supporting Research in Maine
The 91AV Bioanalytical Core supports the research community in the state of Maine (and beyond) through collaborations with research institutions, health care centers, and private companies, including start-ups. The core currently provides bioanalytical expertise for NIH-funded grants focused on drug discovery, basic and translational biology, and pharmacology. Specifically, the core provides in vitro screening for physicochemical properties, pharmacokinetic (PK) analysis and tissue niche drug exposure for diverse classes of molecules, from pre-clinical and clinical studies. The core also provides assay development and quantification of important biomolecules to support proof of concept and mechanistic studies. Additionally, quantification of molecules in environmental samples and field studies is an emerging area of focus for the core.
Developing Maine’s Workforce
Since 2010, the Bioanalytical Core has played a crucial role in preparing young scientists for a future in research in Maine. Mentoring students at the high school level through graduate and medical studies, the Core has introduced basic laboratory skills such as pipetting and solution preparation, along with more advanced analytical methodologies such as sample extraction, ELISA, and LC-MS/MS analysis. The Core has developed a short course on mass spectrometry to introduce students and researchers alike to this widely used industrial and academic research technique. These efforts have provided data and experience for high school Capstone projects, medical research fellowships, INBRE summer research fellows, and graduate student theses (M.S. and Ph.D.).
Work Spaces
91AV’s Bioanalytical Core is housed within the Portland Laboratory for Biotechnology and Health Sciences (PLBHS), a collaborative space for 91AV researchers, graduate students, postdoctoral fellows, and start-ups. Located on the third floor of the School of Pharmacy building, the Core shares a 3,220-square-foot open lab with 22 benches and a 204-square-foot cell culture facility equipped with two biological safety cabinets and incubators.
Key Instrumentation
Shared with the PLBHS, instrumentation includes:
- High-sensitivity LC-MS/MS (Agilent 6460 equipped with 1200 series HPLC)
- Five chemical fume hoods
- -80°C Freezers
- refrigerator/freezers
- -20°C freezers
- Olympus BX51 microscope
- Cytex Guava EasyCyt flow cytometer
- BioRad thermocyclers
- Protein Simple FluorChem M
- Protein Discovery Gel Free 8100 Fractionation System
- Fluoroskan Ascent fluorometer
- Malvern Zetasizer Ultra
- TA instruments Isothermal Titration Calorimeter
- Spectromax plate reader
- Low, moderate and high speed centrifuges
- WinNonLin software for PK analysis
Current Catalog of LC-MS/MS Methods
Biomarkers
A validated supported liquid extraction method with deuterated internal standards; analytes derivatized with dansyl chloride subsequent to extraction. Linear range: 0.100-50.0 nM in 40 µL plasma/serum for testosterone and estradiol.
A validated strong cation exchange solid phase extraction from 20 µL plasma using deuterated internal standards. Linear range from plasma/serum 1.00-1000 nM for dopamine, 2.50-1000 nM for epinephrine, and norepinephrine.
Extraction from pooled zebrafish larvae or DMEM cell culture media, using deuterated internal standards. Linear range: 2.00-500 nM for PGD2 and 20.0-200 nM for PGE2
Drugs
A protein precipitation method for the determination of risperidone and olanzapine from single zebrafish larvae when dosed in tank water. Linear range: 100-10,000 nM for risperidone and paliperidone; 200-10,000 nM for olanzapine.
A validated method for the determination of analytes in 20 µL mouse plasma and bone marrow using paroxetine as an internal standard. Plasma and marrow were prepared by protein precipitation with acetonitrile. Linear range: 0.500-1,000 nM for each analyte in plasma and marrow.
A supported liquid extraction method for quantitation of nimodipine and deuterated internal standard from human plasma and cerebrospinal fluid. Linear range: 2.50-120 nM.
Method developed to quantitate all four analytes in one analytical run, uses deuterated internal standards. Developed for use in determining concentration in drug formulation. No extraction; sample diluted and analyzed by LC-MS/MS. Linear range 50.0-1,000 nM for acetaminophen, leucovorin, and ampicillin. Linear range 5.00-1,000 nM for levetiracetam.
Protein precipitation method from cell lysates; linear range 5-1,000 nM.
Protein precipitation from 20 µL mouse plasma; linear range 1.00-1,000 nM.
Concentrations isolated from mouse plasma, brain, and spinal cord via protein precipitation; linear range 2.00-1,000 nM.
Extracted from 50 µL mouse plasma via protein precipitation. Linear range 1.00-2000 nM.
A validated solid phase extraction method from 20 µL mouse plasma using a deuterated internal standard. Linear range 1.00-1,000 nM.
