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High-Resolution Mass Spectrometry (HRMS)

High-Resolution Mass Spectrometry (HRMS)

The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For...

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Related Experiment Video

Updated: May 6, 2026

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method

Published on: October 7, 2025

Intact microRNA analysis using high resolution mass spectrometry.

Majlinda Kullolli¹, Emily Knouf, Maria Arampatzidou

¹Canary Center for Cancer Early Detection, Department of Radiology, Stanford University School of Medicine, Palo Alto, CA, 94304, USA.

Journal of the American Society for Mass Spectrometry

|November 1, 2013

Summary

High resolution mass spectrometry can separate and measure microRNAs (miRNAs) and their variants. This technique, using liquid chromatography-mass spectrometry, offers a new tool for analyzing these important regulatory molecules.

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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

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Last Updated: May 6, 2026

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method

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Published on: October 7, 2025

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

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Published on: November 18, 2014

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

Area of Science:

Molecular Biology
Biochemistry
Analytical Chemistry

Background:

MicroRNAs (miRNAs) are small non-coding RNAs crucial for gene regulation, cellular processes, and disease.
Understanding miRNA origins and functions requires advanced analytical tools.

Purpose of the Study:

To evaluate high-resolution mass spectrometry for analyzing microRNAs (miRNAs).
To demonstrate the capability of mass spectrometry in separating and quantifying miRNAs and their 3' variants.

Main Methods:

Proof-of-concept experiments using high-resolution mass spectrometry.
Analysis of miRNA separation and mass accuracy.
Comparison of collision-induced dissociation (CID) and higher-energy collisional dissociation (HCD) fragmentation.
Measurement of dynamic range, limit of detection, and limit of quantitation.
Online liquid chromatography-mass spectrometry (LC-MS) with data-dependent acquisition.

Main Results:

Mass spectrometry successfully resolved and separated miRNAs and their 3' variants in mixtures with low ppm mass accuracy.
Both CID and HCD fragmentation provided similar sequence coverage, with HCD offering additional fragmentation.
Linear dynamic range, limit of detection, and limit of quantitation were determined for miRNA analysis.
Online LC-MS enabled fragmentation of multiple charge states, achieving near-full miRNA sequence coverage on a chromatographic timescale.

Conclusions:

High-resolution mass spectrometry is effective for separating and measuring miRNAs in complex mixtures.
Standard LC-MS setups can be adapted for online miRNA analysis, providing valuable insights into their physiology and function.