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Inductively Coupled Plasma-Mass Spectrometry (ICP-MS): Interferences

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Inductively coupled plasma–mass spectrometry (ICP–MS) is a highly selective and sensitive technique for accurate elemental analysis. Though the analysis of ICP–MS mass spectra is comparatively straightforward, it is affected by spectroscopic and non-spectroscopic interferences. Spectroscopic interferences arise when the plasma contains ionic species with an m/z value the same as the analyte ion. Spectroscopic interference can be categorized as isobaric, polyatomic ions, and...

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Related Experiment Video

Updated: Oct 9, 2025

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging cPILOT

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Published on: December 18, 2020

Assessing interference in isobaric tag-based sample multiplexing using an 18-plex interference standard.

Jose Navarrete-Perea¹, Xinyue Liu¹, Ramin Rad¹

¹Department of Cell Biology, Harvard Medical School, Boston, MA, USA.

|December 17, 2021

Summary

This summary is machine-generated.

New Triple yeast KnockOut (TKO) standards and updated software (TVT2.5) help reduce reporter ion interference in isobaric mass spectrometry experiments. Real-time database search (RTS) improves proteome coverage and minimizes interference, offering a versatile platform for method development.

Keywords:

Eclipse FAIMS IFI RTS SPS-MS3 TMTpro18

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Area of Science:

Proteomics
Mass Spectrometry
Analytical Chemistry

Background:

Reporter ion interference is a key challenge in isobaric tag-based proteomics, limiting multiplexing accuracy.
Existing methods like real-time database search (RTS) show promise but require robust benchmarking tools.
There is a continuous need to evaluate sample preparation and data acquisition strategies for interference reduction.

Purpose of the Study:

To introduce updated Triple yeast KnockOut (TKO) standards (TKO18 and TKO18L) for assessing isobaric tag-based experiments.
To evaluate the effectiveness of real-time database search (RTS) and LysC digestion in reducing interference and improving proteome coverage.
To present an upgraded TKO viewing tool (TVT2.5) for analyzing proteomic data.

Main Methods:

Development of TKO18 (TMTpro18-plex) and TKO18L (LysC-digested TKO18) standards.
Acquisition of proteomic data using different acquisition modes.
Analysis of data using the updated TKO viewing tool (TVT2.5) with and without RTS.
Comparison of proteome coverage and interference levels between TKO18 and TKO18L.

Main Results:

Real-time database search (RTS) significantly reduces reporter ion interference and enhances proteome coverage.
LysC-only digestion offers a modest reduction in interference but comes at the cost of reduced proteome depth.
The TKO18 and TKO18L standards, analyzed with TVT2.5, provide a comprehensive platform for method evaluation.

Conclusions:

The updated TKO standards and TVT2.5 software offer a valuable resource for developing and validating methods to mitigate interference in isobaric tag-based proteomics.
RTS is an effective strategy for improving data quality in multiplexed quantitative proteomics.
Optimizing sample preparation and data acquisition is crucial for maximizing proteome coverage and accuracy.