{"id":943,"date":"2014-06-18T08:05:39","date_gmt":"2014-06-18T15:05:39","guid":{"rendered":"http:\/\/sop.washington.edu\/?page_id=943"},"modified":"2025-04-30T08:54:37","modified_gmt":"2025-04-30T15:54:37","slug":"services","status":"publish","type":"page","link":"https:\/\/staff.washington.edu\/twentzel\/wordpress\/department-of-medicinal-chemistry\/research\/mass-spectrometry-center\/services\/","title":{"rendered":"Services"},"content":{"rendered":"

Recognizing the diversity of research interests on- and off-campus, the Center offers the following three \u201cfor fee\u201d services to the scientific community: Submitted Sample Analysis; Investigator Instrument Use; and Protocol Development. A breakdown of charges for services under each category may be found under\u00a0Rates.<\/a><\/p>\n

Submitted Sample Analysis<\/h3>\n

Listed below are the types of analyses routinely performed by Center personnel on submitted samples. A rate schedule for these analyses may be found in\u00a0Rates under Submitted Sample Analysis<\/a>. Sample preparation and submission requirements are discussed in\u00a0Sample Submission<\/a>. Please contact the Center prior to submission of samples requiring \u201cspecial\u201d handling or analysis.<\/p>\n

\n\n\n\n\n\n\n\n\n\n\n
TYPE<\/td>\nDESCRIPTION<\/td>\n<\/tr>\n
I<\/td>\nLR (MALDI, ESI & APcI; \u00b1 eV)<\/strong>
\nLow Resolution (LR unit mass) analysis of gases, liquids and solids by an appropriate sample inlet system<\/td>\n<\/tr>\n
II<\/td>\nHR (MALDI, ESI & APcI; \u00b1 eV)<\/strong>
\nHigh Resolution accurate mass measurement (AMM) coupled with empirical formula or amino acid determination; mass range < 2000 Dalton; Resolving Power (RP) 5000 \u2013 > 400,000.<\/td>\n<\/tr>\n
III<\/td>\nLC-MS, LR (ESI & APcI; \u00b1 eV)<\/strong>
\nLiquid Chromatographic (LC) separation combined with LR mass spectral analysis; mass range < 3000 Dalton<\/td>\n<\/tr>\n
IV<\/td>\nLR LC-MS and LC-MS\/MS Quantification and\/or Detection<\/strong>
\nLC separation coupled with the selected ion monitoring (SIM), or selected and multiple reaction monitoring techniques, SRM and MRM, respectively, using tandem mass analyzers (MS\/MS).<\/td>\n<\/tr>\n
V<\/td>\nHR LC-MS\/MS Structure Elucidation and\/or Component Scouting<\/strong>
\nUse of collisional induced dissociation (CID) in product (daughter) ion, precursor (parent) ion or neutral loss scanning analysis.<\/td>\n<\/tr>\n
VI<\/td>\nIntact Protein and Peptide Molecular Weight<\/strong>
\nMultiple charge state ESI spectra of a biopolymer(s) are deconvoluted to determine the zero charge mass of the component(s). (Depending on presence of salts, detergents or other ionic impurities in the sample, on-line LC-MS or other off-line cleanup procedures may be required.)<\/td>\n<\/tr>\n
VII<\/td>\nPeptide Mapping and Proteomics Studies<\/strong><\/p>\n

Sequence coverage and modification ID.\u00a0 Protein samples are cleaned up using s-trap spin columns and trypsin digested.\u00a0 Depending\u00a0on samples either nanoflow or microflow LC conditions will be applied.<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n

See Ionization Methods and Operational Modes<\/a> for more information relating Center instrumentation and capabilities.<\/p>\n

Investigator Instrument Use<\/h3>\n

\u201cHands-on\u201d instruction in the use of all Center instruments is available on all of our equipment. The level of instruction is such that following the completion of training (usually 2 to 4 hrs) and several periods of supervised use, the researcher may exercise independent unattended use of an instrument with instructor approval. Instruction is instrument specific and is provided at the discretion of the management based on anticipated project or research group needs. Charges for instruction and investigator use may be found under Investigator Instrument Use.<\/p>\n

Protocol Development<\/h3>\n

The Center offers both Sample Preparation and Mass Spectrometry Assay protocol development services. Dependent upon the need, assay protocol development may include: a methodology appropriate for sample preparation prior to MS analysis; a methodology for on-line LC-MS separations, if necessary; the running of preliminary Standard Curves, Reproducibility Studies; and the determination of Limits of Detection (LOD). The transfer of the final protocol to project personnel may also be included, however, complete validation of the method is not.<\/p>\n

Protocol development services are provided at the discretion of the management and are based, in part, on the expectation that Center instrumentation will be used in carrying out subsequent studies. For information on the costs associated with this service, please check Rates under Protocol Development or contact\u00a0Dale Whittington<\/a>\u00a0of our Staff.<\/p>\n

The Center is not equipped or licensed to handle radioisotopes and, consequently, will NOT work with radioactive materials. However, the Center will consult on sample preparations involving these media.<\/p>\n

Quantitative Proteomics<\/h3>\n

We apply LC-MS\/MS proteomics (MRM) for the quantification of proteins in complex biological systems, e.g., tissues, blood, urine and cerebrospinal fluid (CSF). The advantages of MRM proteomics are: i) selectivity, i.e., it can distinguish highly homologous proteins; ii) quantification of multiple proteins in a short time (approximately 10 proteins in 20 min) with almost similar or better sensitivity than western blotting; iii) antibodies are not required for quantification; iv) it offers absolute protein expression, and; v) data quality is better than western blotting as MRM relies on multiple signals, i.e., multiple peptides and multiple MRM transitions.<\/p>\n

We offer research support to projects such as 1) targeted quantification of proteins (in tissues and biofluids) to investigate disease pathophysiology, and 2) discovery\/validation of hypothesis driven biomarkers of diseases\/drug efficacy and toxicity.<\/p>\n

Ionization Methods and Operational Modes<\/h3>\n

Positive and negative ions may be generated in all modes of ionization. The choice of an appropriate ionization mode and its polarity is dependent on the nature of the compound of interest (functional groups, volatility, stability, etc.) and on the method of sample introduction amenable to it.<\/p>\n

In addition, the mass analyzer (time-of-flight, quadrupole, etc.) used in conjunction with a particular ionization and sample inlet technique will set limits on the resolution and mass range of the analysis. The following table attempts to give an overview of useful combinations based on the instrumentation available in the Center.<\/p>\n

\n\n\n\n\n\n\n\n\n\n
Ionization Mode<\/strong><\/td>\nESI<\/strong><\/td>\nAPcl<\/strong><\/td>\nMALDI<\/strong><\/td>\n<\/tr>\n
Mass Range Daltons (Da)<\/td>\n50 \u2013 > 6,000<\/td>\n50 \u2013 6,000<\/td>\n500 \u2013 100,000<\/td>\n<\/tr>\n
Analyte Polarity<\/td>\nMedium \u2013 Very High<\/td>\nMedium \u2013 High<\/td>\nLow \u2013 Very High<\/td>\n<\/tr>\n
Ion Charge electron Volts (eV)<\/td>\n\u00b1<\/td>\n\u00b1<\/td>\n\u00b1<\/td>\n<\/tr>\n
LC-MS<\/td>\nYes<\/td>\nYes<\/td>\nYes<\/td>\n<\/tr>\n
HR High Resolution<\/td>\nYes<\/td>\nYes<\/td>\nYes<\/td>\n<\/tr>\n
CID Collision Induced Dissociation<\/td>\nYes<\/td>\nYes<\/td>\nYes<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n","protected":false},"excerpt":{"rendered":"

Recognizing the diversity of research interests on- and off-campus, the Center offers the following three \u201cfor fee\u201d services to the scientific community: Submitted Sample Analysis; Investigator Instrument Use; and Protocol Development. 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