Strategies and Analytical Workflows to Extend the Dynamic Range in Quantitative LC–MS/MS Analysis

Ediage, Emmanuel Njumbe; Aerts, Tania; Lubin, Arnaud; Cuyckens, Filip; Dillen, Lieve; Verhaeghe, Tom

doi:10.4155/bio-2018-0309

Cited by 6 publications

(3 citation statements)

References 19 publications

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“…To avoid detector saturation (an issue seen at the highest concentrations of HMP), only 5 μL of the highest calibrators was injected. This allows to extend the calibration range, 28 rendering it possible to quantify potentially very high concentrations in the genetically modified lines. For all analytes, a bias below 12.5% was found for all concentration levels.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

The First Comprehensive LC–MS/MS Method Allowing Dissection of the Thiamine Pathway in Plants

et al. 2020

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Arabidopsis thaliana serves as a model plant for genetic research, including vitamin research. When aiming at engineering the thiamine (vitamin B1) pathway in plants, the availability of tools that allow the quantitative determination of different intermediates in the biosynthesis pathway is of pivotal importance. This is a challenge, given the nature of the compounds and the minute quantities of genetically engineered material that may be available for analysis. Here, we report on the first LC–MS/MS method for the simultaneous quantification of thiamine, its mono- and diphosphate derivatives and its precursors 4-methyl-5-(2-hydroxyethyl) thiazole (HET) and 4-amino-2-methyl-5-hydroxymethylpyrimidine (HMP). This method was optimized and validated for the quantitative determination of these analytes in Arabidopsis thaliana. All analytes were chromatographically separated within less than 2.5 min during an 8 min run. No unacceptable interferences were found. The method was fully validated based on international guidelines. Accuracy (%bias) and total imprecision (%CV) were within preset acceptance criteria for all analytes in both QC and real samples. All analytes were stable in extracted samples when stored for 48 h at 4 °C (autosampler stability) and when reanalyzed after storage at −80 °C and −20 °C for 2 weeks (freeze/thaw stability). We demonstrated the start material should be stored at −80 °C to ensure stability of all analytes during short- and long-term storage (up to 3 months). The validity and applicability of the developed procedure was demonstrated via its successful application on Arabidopsis lines, genetically engineered to enhance thiamine content.

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Section: ■ Results and Discussionmentioning

confidence: 99%

The First Comprehensive LC–MS/MS Method Allowing Dissection of the Thiamine Pathway in Plants

et al. 2020

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“…This process can be further enhanced and automated with the deployment of imaging tools and imaging analysis software packages. In addition, further insight may be gained from future studies if the analytical approach included the use of LC-MS/MS (46). LC-MS/MS is highly sensitive and selective and can provide insight into the ionization state which may not be possible with the chromatography approach presented here (47).…”

Section: Discussionmentioning

confidence: 99%

Automated High Throughput pKa and Distribution Coefficient Measurements of Pharmaceutical Compounds for the SAMPL8 Blind Prediction Challenge

Bahr

Nandkeolyar

Kenna

et al. 2021

Preprint

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The goal of the SAMPL (Statistical Assessment of the Modeling of Proteins and Ligands) challenge is to improve the accuracy of current computational models to estimate free energy of binding, deprotonation, distribution and other associated physical properties that are useful for the design of new pharmaceutical products. New experimental datasets of physicochemical properties provide opportunities for prospective evaluation of computational prediction methods. Here, aqueous pKa and a range of bi-phasic logD values for a variety of pharmaceutical compounds were determined through a streamlined automated process to be utilized in the SAMPL8 physical property challenge. The goal of this paper is to provide an in-depth review of the experimental methods utilized to create a comprehensive data set for the blind prediction challenge. The significance of this work involves the use of high throughput experimentation equipment and instrumentation to produce acid dissociation constants for twenty-three drug molecules, as well as distribution coefficients for eleven of those molecules.

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“…As a result, tissue samples may contain NP concentrations spanning 6 to 10 orders of magnitude (23). This makes quantitative studies challenging because endogenous NP levels, typically between 3 to 5 orders of magnitude, may fall outside of the linear dynamic range of the mass spectrometer (24). Wide NP concentrations within a sample are problematic for MS quantitation, as these could lead to spacecharge effects, which reduce the linear intra-scan dynamic range of the instrument, effectively masking the signal from trace NPs.…”

Section: Biological Factors Influencing Quantitationmentioning

confidence: 99%

Mass Spectrometry Measurements of Neuropeptides: From Identification to Quantitation

Toba

Bell

Romanova

et al. 2022

Annual Rev. Anal. Chem.

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Neuropeptides (NPs), a unique class of neuronal signaling molecules, participate in a variety of physiological processes and diseases. Quantitative measurements of NPs provide valuable information regarding how these molecules are differentially regulated in a multitude of neurological, metabolic, and mental disorders. Mass spectrometry (MS) has evolved to become a powerful technique for measuring trace levels of NPs in complex biological tissues and individual cells using both targeted and exploratory approaches. There are inherent challenges to measuring NPs, including their wide endogenous concentration range, transport and postmortem degradation, complex sample matrices, and statistical processing of MS data required for accurate NP quantitation. This review highlights techniques developed to address these challenges and presents an overview of quantitative MS-based measurement approaches for NPs, including the incorporation of separation methods for high-throughput analysis, MS imaging for spatial measurements, and methods for NP quantitation in single neurons. Expected final online publication date for the Annual Review of Analytical Chemistry Volume 15 is June 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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Strategies and Analytical Workflows to Extend the Dynamic Range in Quantitative LC–MS/MS Analysis

Cited by 6 publications

References 19 publications

The First Comprehensive LC–MS/MS Method Allowing Dissection of the Thiamine Pathway in Plants

The First Comprehensive LC–MS/MS Method Allowing Dissection of the Thiamine Pathway in Plants

Automated High Throughput pKa and Distribution Coefficient Measurements of Pharmaceutical Compounds for the SAMPL8 Blind Prediction Challenge

Mass Spectrometry Measurements of Neuropeptides: From Identification to Quantitation

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