The NJ Mass Spectrometry Discussion Group presents its June meeting on Tuesday, June 14, 2016 at at the Holiday Inn Somerset-Bridgewater, 195 Davidson Ave, Somerset NJ 08873 [ hotel website ]
Sponsored by Shimadzu
The evening is free for attendees, courtesy of our sponsor!
Please register here.
5:30 pm Social and registration
6:15 pm Complimentary dinner
7:00 pm Welcome and opening remarks
7:05 pm Speakers
I. Evelyn Wang, PhD Candidate, August 2016
University of Texas at Arlington
Intact Protein quantitation using a Triple Quadrapole Mass Spectrometer
The demand is increasing for protein detection and quantitation in biological fluids for disease detection, protein therapeutics monitoring, and drug development. Current methods use highly sensitive and specific triple quadrupole mass spectrometry (QqQ-MS) to quantify protein-digested peptides to then correlate original intact protein concentrations in the sample. This bottom-up method for protein quantitation can introduce errors. Therefore, for more accurate protein quantitation researchers use expensive isotopically labeled proteins for standards. A method that bypasses the protein digestion step to directly quantify intact proteins on QqQ-MS was developed and will be presented. Myoglobin, cytochrome c, lactalbumin, lysozyme, and ubiquitin were selected as protein standards for the proof-of-principle work. Our intact protein quantitation method was developed on a Shimadzu LCMS-8050 QqQ-MS utilizing multiple reaction monitoring (MRM). The experimental pathway and associated challenges that ultimately led to MRM transitions for all protein standards will be shown. The result was calibration curves of respectable linearity (R2>0.99). Further, in order to address complex matrices in biological fluids for future applications, a generic reversed-phase chromatography method was developed on Restek Wide Pore Viva C4, C8, C18, Biphenyl, and PFP Propyl (2.1 x 100 mm; 5 ?m; 300 Å) columns. Prostate specific antigen (PSA) was also included in the study to prove the feasibility of the method for both the liquid valtvalacyc.com chromatography and mass spectrometry aspects. Specificity of the MRM detection was evaluated for both urine and plasma matrices. The method is envisioned to be a model for future development of targeted methods for analysis of important disease indicators such as proteins in biological fluids, especially for clinical diagnostic and treatment advancements.
II. Scott A. Kuzdzal, PhD
General Manager of Marketing, Shimadzu Scientific Instruments
Fully Integrated & Automated LC-MS/MS Sample Preparation and a Brief Introduction to other New Innovations from Shimadzu
In clinical and pharmaceutical laboratories, time is money. Sample preparation for the detection of target analytes such as steroids and immunosuppressant drugs in serum by LC-MS/MS typically involves complex, offline extraction methods such as solid phase extraction or liquid/liquid extraction, both of which require additional sample concentration and reconstitution in appropriate solvents. These sample preparation methods are time-consuming, often taking one hour or more per sample, and are more vulnerable to variability due to errors in manual preparation. We herein present the full integration and automation of steroid and immunosuppressant assay sample preparation and LC-MS/MS analysis using an innovative, automated sample preparation instrument (CLAM-2000) coupled directly to Shimadzu ultra-fast mass spectrometry. This completely integrated, automated quantification method for immunosuppressant and steroid drug compounds allows routine analysis with high data quality/precision, reduced time, increased throughput and enhanced safety.
This talk will also introduce a new ultra-high speed LCMS system for multiplexed analyses that allows users to double the throughput of an existing method. In an analysis of four biomarker compounds for the four major molecular species in the Cytochrome P450 family, the innovative, new Nexera-MX system completed the analysis in only 38 seconds whereas conventional HPLC required one minute and twenty-two seconds.