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Standard Addition as a Method for Quantitative Mass Spectrometry Imaging.

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Quantifying analytes in mass spectrometry imaging (MSI) is challenging. This study introduces a novel standard addition method using tissue extracts for cost-effective and versatile pixel-by-pixel quantification, overcoming limitations of internal standards.

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Area of Science:

  • Analytical Chemistry
  • Biomedical Imaging

Background:

  • Quantification in mass spectrometry imaging (MSI) is difficult due to complex sample matrices.
  • Isotopically labeled internal standards (IS) are commonly used for relative quantification but have limitations like preselection needs and cost.
  • Alternative methods are needed for accurate and accessible MSI quantification.

Purpose of the Study:

  • To develop and validate a novel quantification workflow for MSI using the standard addition method.
  • To compare the performance of standard addition with existing methods like IS quantification and external calibration.
  • To demonstrate the feasibility of using endogenous tissue molecules as calibration standards for standard addition in MSI.

Main Methods:

  • Developed a workflow for acquiring and processing quantitative data using the standard addition method in MSI.
  • Applied standard addition to MSI experiments, utilizing tissue extracts as calibration standards.
  • Compared quantification results from standard addition against IS quantification and external calibration methods.

Main Results:

  • The standard addition method provides a viable approach for pixel-by-pixel quantification in MSI.
  • Quantification results obtained by standard addition were compared with those from IS quantification and external calibration.
  • Demonstrated the successful application of endogenous tissue molecules as readily accessible standards for standard addition quantification.

Conclusions:

  • The proposed standard addition method offers a cost-effective, accessible, and versatile alternative for MSI quantification.
  • This approach overcomes the limitations associated with traditional methods, such as the cost and availability of isotopically labeled IS.
  • The use of endogenous analytes as calibration standards enhances the practicality and applicability of standard addition in MSI.