The Pharmacoanalytical Shared Resource supports pre-clinical and clinical drug development at OSUCCC–James by providing high quality, cost-effective and reliable bioanalytical method development, quantitative sample analysis, and pharmacokinetic/pharmacodynamic/pharmacogenetic experimental design and data analysis.
Primary PhASR equipment comprises three liquid chromatography/mass spectrometry (LC-MS) systems, which include a quadrupole time-of-flight (Q-tof) and two triple-quadrupole mass spectrometers. One triple quadrupole instrument is supported by an ultra-high pressure liquid chromatography (UHPLC) system designed for high-throughput sample analysis, and the other two are supported by high pressure liquid chromatography (HPLC) systems.
Liquid Chromatography/Mass Spectrometry (LC-MS) System Details:
Thermo Fisher TSQ Quantiva: a triple quadrupole mass spectrometer with Ion Max NG™ ionsource and EASY-Spray NG™ ionsource (nanospray) is connected to a Thermo Dionex UltiMate 3000 RSLCnano System (degasser, binary nano pump, ternary microflow pump, column heater and autosampler). The Quantiva system has attogram sensitivity and is the most powerful triple quadrupole instrument on the market. Using the "Active Ion Management" technology the Quantiva system provides high sensitivity and high resolution for both Q1/Q3 full scan and selected-reaction monitoring (SRM) with the mass range of m/z 10-1850. It can perform 15,000 amu/s full scan at a resolution of 2 FWHM and 500 SRMs/s with up to 30,000 definable SRMs. The Quantiva is coupled with an Ultimate 3000 RSLC nano system enabling robust HPLC capabilities for the separation of complex matrices. This system went on-line in 2014, and is utilized primarily for quantitative protein/peptide assays and for methods requiring high sensitivity, such as accurate quantification in low quantity samples (e.g. fine needle biopsies).
Thermo Finnigan TSQ Quantum Ultra: a triple quadrupole mass spectrometer with extended mass range (EMR) and electrospray ionization (ESI) probe is connected to a Thermo Dionex UltiMate 3000 RSLCnano System (degasser, binary nano pump, ternary microflow pump, column heater and autosampler). The TSQ Quantum Ultra EMR is capable of quantifying charged molecules with a mass/charge ratio (m/z) up to 3000 at high resolution (0.2 m/z mass peak width) and high sensitivity. The EMR capability allows improved high-resolution analysis of larger biomoleculescompared to the standard m/z limit of 1500. The Ultimate 3000 RSLC nano system enables robust separation of complex matrices andcan be operated at capillary or micro flowrates as necessary. This system went on-line in 2006, and is utilized primarily for method development and secondarily for sample analysis.
Thermo Finnigan TSQ Quantum Discovery Max: a triple quadrupole mass spectrometer with ESI and atmospheric pressure chemical ionization (APCI) probes is coupled with a Thermo Accela UHPLC system (quaternary pumps, degasser, column heater, PDA, and a CTC autosampler with temperature control and expandable multiplexing capabilities). The TSQ Quantum Discovery is capable of quantifying molecules with a mass/charge ratio up to 1500 with high resolution (0.2 m/z mass peak width) and moderate to high sensitivity. The APCI probe is interchangeable with the ESI probes on both the Discovery and Ultra systems which provide alternative ionization strategies for large molecules and for molecules with low ionization potential. Importantly, the Accela UHPLC with CTC autosampler is expandable for online sample extraction and multiplexing that would enable automated method development and simultaneous runs using multiple methods. This LC-MS system went on-line 2005 and is used primarily for sample analysis and secondarily for method development.
Thermo Finnigan LCQ Deca: An ion trap mass spectrometer with Ion Max API Source connected to a Shimadzu Prominence LC20 system (degasser, binary pumps, mobile phase switching valve, column oven, autosampler and controller). The LCQ Deca system has improved ion optics for enhanced sensitivity and greater precision with mass range up to m/z 2000. It is particularly useful for qualitative structural characterization due to its ability to perform multiple stage (MSn) ion fragmentation analyses (up to 10 stages). This system provides reliable and precise metabolite identification, peptide mapping, and complex mixture analysis. The Shimadzu Prominence LC system enables robust HPLC performance and flexibility This system went on-line in 2005 and is utilized primarily for metabolite identification and secondarily for nucleotide/deoxynucleotide triphosphates analysis.
Waters/Micromass Q-Tof Micro hybrid: A quadrupole/time of flight hybrid mass spectrometer with lockspray, ESI and APCI probes coupled with a Waters 2795 Alliance HT HPLC containing a ternary pump, temperature controlled column compartment and temperature controlled autosampler. This system enables accurate mass determination of small molecules and supports metabolite identification. A hexapole collision cell also provides the enhanced structural investigations such as evaluation of metabolites and degradation products.
PhASR also maintains full access to other resources necessary to develop and run biological correlative assays, including UV-Vis and fluorescence spectrometers, plate-readers, thermal cyclers, centrifuges, cell culture and animal facilities, and freezer and refrigerator storage.
WinNonlin and NONMEM Pharmacokinetic Analysis Software Packages are also available for data analysis.