scholarly journals Single Quadrupole Multiple Reaction Monitoring Q-MRM enables Quantitative Mass Spectrometry

2021 ◽  
Author(s):  
Jingchuan Xue ◽  
Rico J.E. Derks ◽  
William Wwebb ◽  
Aries Aisporna ◽  
Martin Giera ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Quantitative Analysis ◽  
Matrix Effects ◽  
Multiple Reaction Monitoring ◽  
Mass Spectrometry Analysis ◽  
Good Precision ◽  
Reaction Monitoring ◽  
Font Size ◽  
Quantitative Mass Spectrometry ◽  
Fragment Ions

<p> </p><p>A single quadrupole combined with enhanced in-source fragmentation/annotation (EISA) was used to perform multiple reaction monitoring (Q-MRM) for quantitative mass spectrometry analysis. EISA amplifies fragmentation of traditional soft electrospray ionization to produce fragment ions that have been found to be identical to those generated in tandem mass spectrometry. We have combined EISA fragmentation data with criteria established by the European Union Commission Directive 2002/657/EC for electron ionization single quadrupole quantitative analysis to perform quantitative Q-MRM experiments. These experiments were performed on multiple types of complex samples that included a mixture of 50 standards, as well as cell and plasma extracts. The dynamic range for Q-MRM quantitative analysis was comparable to triple quadrupole multiple reaction monitoring (QqQ-MRM) analyses at up to 5 orders of magnitude with the cell and plasma extracts showing similar matrix effects across both platforms. Amino acid and fatty acid measurements performed from certified NIST 1950 plasma with isotopically labelled standards demonstrated Q-MRM accuracy in the range of 91-110% for the amino acids, 76-129% for the fatty acids, and good precision (coefficient of variation < 10%). In order to enhance specificity, a newly developed Correlated Ion Monitoring (CIM) algorithm was designed to facilitate these analyses. CIM autonomously processes, aligns, filters, and compiles multiple ions within one chromatogram enabling both precursor and in-source fragment ions to be correlated within a single chromatogram, also enabling the detection of co-eluting species based on precursor and fragment ion ratios. Q-MRM and CIM with single quadrupole instrumentation has been designed as an alternative to QqQ-MRM technology by correlating precursor and fragment ions to facilitate high sensitivity Q-MRM quantitative analysis on existing instrumentation that are generally inexpensive, easy to operate, and technically less complex. </p> <p>@font-face {font-family:"Cambria Math"; panose-1:2 4 5 3 5 4 6 3 2 4; mso-font-charset:0; mso-generic-font-family:roman; mso-font-pitch:variable; mso-font-signature:3 0 0 0 1 0;}p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-unhide:no; mso-style-qformat:yes; mso-style-parent:""; margin:0in; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Times New Roman",serif; mso-fareast-font-family:"Times New Roman";}.MsoChpDefault {mso-style-type:export-only; mso-default-props:yes; font-size:11.0pt; mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:DengXian; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi; mso-fareast-language:ZH-CN;}.MsoPapDefault {mso-style-type:export-only; margin-bottom:8.0pt; line-height:107%;}div.WordSection1 {page:WordSection1;}</p>

scholarly journals EISA-MRM: Multiple Reaction Monitoring with Single Quadrupole Mass Spectrometry

2021 ◽  
Author(s):  
Jingchuan Xue ◽  
Rico J.E. Derks ◽  
William Wwebb ◽  
Aries Aisporna ◽  
Martin Giera ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Quantitative Analysis ◽  
Dynamic Range ◽  
Matrix Effects ◽  
Multiple Reaction Monitoring ◽  
Mass Spectrometry Analysis ◽  
Reaction Monitoring ◽  
Quadrupole Mass ◽  
Fragment Ions ◽  
Specificity And Sensitivity

<p>Enhanced in-source fragmentation annotation combined with single quadrupole multiple reaction monitoring (EISA-MRM) has been designed for quantitative mass spectrometry analysis. EISA contrasts to traditional electrospray as a soft ionization technology and is proving to be advantageous since the resulting fragment ions are identical as those generated in tandem mass spectrometry. Criteria established by the European Union Commission Directive 2002/657/EC for electron ionization single quadrupole quantitative analysis was used for the EISA-MRM quantitative analyses and experiments were performed on multiple types of complex samples that included a mixture of 50 standards, as well as cell and plasma extracts. The dynamic range for quantitative analysis was comparable to QqQ-MRM analyses at up to 5 orders of magnitude and the EISA-MRM and QqQ-MRM of the cell and plasma extracts showed similar matrix effects. Amino acid and fatty acid measurements performed from certified NIST 1950 plasma with isotopically labelled standards demonstrated EISA-MRM accuracy in the range of 91-110% for the amino acids, 76-129% for the fatty acids, and precision with a CV < 10%. In order to enhance specificity and sensitivity, a newly developed Correlated SIM Chromatogram (CSC) algorithm was designed to facilitate MRM quality analyses. The EISA-MRM quantitative analysis with CSC informatics enables both precursor and in-source fragment ions to be correlated within a single quadrupole mass spectrometer.</p>

scholarly journals EISA-MRM: Multiple Reaction Monitoring with Single Quadrupole Mass Spectrometry

2021 ◽  
Author(s):  
Jingchuan Xue ◽  
Rico J.E. Derks ◽  
William Wwebb ◽  
Aries Aisporna ◽  
Martin Giera ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Quantitative Analysis ◽  
Dynamic Range ◽  
Matrix Effects ◽  
Multiple Reaction Monitoring ◽  
Mass Spectrometry Analysis ◽  
Reaction Monitoring ◽  
Quadrupole Mass ◽  
Fragment Ions ◽  
Specificity And Sensitivity

<p>Enhanced in-source fragmentation annotation combined with single quadrupole multiple reaction monitoring (EISA-MRM) has been designed for quantitative mass spectrometry analysis. EISA contrasts to traditional electrospray as a soft ionization technology and is proving to be advantageous since the resulting fragment ions are identical as those generated in tandem mass spectrometry. Criteria established by the European Union Commission Directive 2002/657/EC for electron ionization single quadrupole quantitative analysis was used for the EISA-MRM quantitative analyses and experiments were performed on multiple types of complex samples that included a mixture of 50 standards, as well as cell and plasma extracts. The dynamic range for quantitative analysis was comparable to QqQ-MRM analyses at up to 5 orders of magnitude and the EISA-MRM and QqQ-MRM of the cell and plasma extracts showed similar matrix effects. Amino acid and fatty acid measurements performed from certified NIST 1950 plasma with isotopically labelled standards demonstrated EISA-MRM accuracy in the range of 91-110% for the amino acids, 76-129% for the fatty acids, and precision with a CV < 10%. In order to enhance specificity and sensitivity, a newly developed Correlated SIM Chromatogram (CSC) algorithm was designed to facilitate MRM quality analyses. The EISA-MRM quantitative analysis with CSC informatics enables both precursor and in-source fragment ions to be correlated within a single quadrupole mass spectrometer.</p>

scholarly journals Precision of Multiple Reaction Monitoring Mass Spectrometry Analysis of Formalin-Fixed, Paraffin-Embedded Tissue

2012 ◽  
Vol 11 (6) ◽  
pp. 3498-3505 ◽  
Author(s):  
Robert W. Sprung ◽  
Misti A. Martinez ◽  
Kristen L. Carpenter ◽  
Amy-Joan L. Ham ◽  
Mary Kay Washington ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Multiple Reaction Monitoring ◽  
Mass Spectrometry Analysis ◽  
Reaction Monitoring ◽  
Spectrometry Analysis ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

scholarly journals Calcineurin Activity Assay Measurement by Liquid Chromatography–Tandem Mass Spectrometry in the Multiple Reaction Monitoring Mode

2014 ◽  
Vol 60 (2) ◽  
pp. 353-360 ◽  
Author(s):  
Lynn Carr ◽  
Anne-Laure Gagez ◽  
Marie Essig ◽  
François-Ludovic Sauvage ◽  
Pierre Marquet ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Mononuclear Cells ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Reaction Monitoring ◽  
Activity Assay ◽  
Peripheral Blood Mononuclear ◽  
Blood Concentrations ◽  
Transplant Patients

Abstract BACKGROUND Blood concentrations of the calcineurin inhibitors (CNIs) cyclosporine and tacrolimus are currently measured to monitor immunosuppression in transplant patients. The measurement of calcineurin (CN) phosphatase activity has been proposed as a complementary pharmacodynamic approach. However, determining CN activity with current methods is not practical. We developed a new method amenable to routine use. METHODS Using liquid chromatography–multiple reaction monitoring mass spectrometry (LC-MRM-MS), we quantified CN activity by measuring the dephosphorylation of a synthetic phosphopeptide substrate. A stable isotope analog of the product peptide served as internal standard, and a novel inhibitor cocktail minimized dephosphorylation by other major serine/threonine phosphatases. The assay was used to determine CN activity in peripheral blood mononuclear cells (PBMCs) isolated from 20 CNI-treated kidney transplant patients and 9 healthy volunteers. RESULTS Linearity was observed from 0.16 to 2.5 μmol/L of product peptide, with accuracy in the 15% tolerance range. Intraassay and interassay recoveries were 100.6 (9.6) and 100 (7.5), respectively. Michaelis–Menten kinetics for purified CN were Km = 10.7 (1.6) μmol/L, Vmax = 2.8 (0.3) μmol/min · mg, and for Jurkat lysate, Km = 182.2 (118.0) μmol/L, Vmax = 0.013 (0.006) μmol/min · mg. PBMC CN activity was successfully measured in a single tube with an inhibitor cocktail. CONCLUSIONS Because LC-MRM-MS is commonly used in routine clinical dosage of drugs, this CN activity assay could be applied, with parallel blood drug concentration monitoring, to a large panel of patients to reevaluate the validity of PBMC CN activity monitoring.

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