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Multicenter Validation Study of Quantitative Imaging Mass Spectrometry.

Jeremy A Barry1, Rima Ait-Belkacem2, William M Hardesty1

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Summary
This summary is machine-generated.

Quantitative imaging mass spectrometry (IMS) shows promising precision and accuracy for drug analysis in rat liver tissue. Stable-isotopically labeled standards improved quantification, especially in dilution series experiments.

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Area of Science:

  • Pharmacology
  • Analytical Chemistry
  • Biomedical Imaging

Background:

  • Quantitative imaging mass spectrometry (IMS) is a powerful tool for analyzing drug distribution in tissues.
  • Variability across sites, analysts, and instruments can hinder the reliability of IMS quantification.
  • Standardization of protocols is crucial for reproducible IMS results.

Purpose of the Study:

  • To evaluate sources of variability in quantitative IMS across different sites, analysts, and instruments.
  • To assess the accuracy and precision of two common IMS quantification methods: mimetic tissue model and dilution series.
  • To investigate the utility of stable-isotopically labeled (SIL) standards for improving IMS quantification and reducing image variability.

Main Methods:

  • Rat liver samples perfused with clozapine were analyzed at three sites by three analysts using a standardized protocol.
  • Clozapine and norclozapine were quantified using mimetic tissue and dilution series methods.
  • Quantification accuracy and precision were compared against liquid chromatography-tandem mass spectrometry (LC-MS/MS).
  • The impact of SIL normalization on precision, accuracy, and image quality was assessed.

Main Results:

  • Both quantitative IMS methods achieved low teens relative standard deviations and approximately 80% accuracy compared to LC-MS/MS.
  • SIL normalization significantly improved accuracy and precision in the dilution series method.
  • Optimal SIL normalization occurred when the analyte-to-internal standard intensity ratio was near one.
  • SIL normalization showed less impact on the mimetic tissue model.

Conclusions:

  • Quantitative IMS can achieve reliable drug quantification in rat liver tissue with standardized protocols.
  • SIL normalization is a valuable tool for enhancing the accuracy and precision of IMS, particularly for the dilution series method.
  • The effectiveness of SIL normalization is dependent on the analyte-to-internal standard ratio, limiting its universal application.