What is the principle of high-resolution mass spectrometry?

High-Resolution Mass Spectrometry (HRMS) is an analytical technique capable of accurately determining ion mass with extremely high mass resolution. It is commonly used for analyzing trace components in complex biological samples and is widely applied in fields such as proteomics, metabolomics, drug metabolism, and biomarker discovery. Its core advantage lies in its ability to accurately measure ion mass, distinguish very close mass differences, and thus achieve high sensitivity and specificity detection of compounds in complex mixtures.

1. Core Principles of High-Resolution Mass Spectrometry: From Ionization to High-Precision Measurement

The principle of high-resolution mass spectrometry is essentially to perform high-resolution determination of the mass-to-charge ratio (m/z) of ions in an ultra-high precision mass analyzer. The process mainly includes the following key steps:

1. Ionization

Using soft ionization techniques, such as ESI (Electrospray Ionization) or MALDI (Matrix-Assisted Laser Desorption/Ionization), molecules in the sample are converted into charged particles. Soft ionization methods can prevent sample fragmentation, maintaining the intact structure of molecules, suitable for the analysis of large molecules (such as proteins, peptides, metabolites, etc.).

2. Ion Transmission and Focusing

The generated ions are transmitted to the mass analyzer under the guidance of an electric field. During this process, ion lenses or quadrupole systems are used to focus the ions' energy, enhancing analysis efficiency and resolution ability.

3. Mass Analyzer: The Core Device of High-Resolution

Currently, mainstream high-resolution mass spectrometers are mainly based on the following three types of mass analyzers:

(1) Orbitrap

• Principle: Ions are captured in a electrostatic potential trap, revolving around the central electrode and oscillating axially. The oscillation frequency is inversely proportional to m/z.
• Features: No magnetic field required, compact structure, resolution up to 100,000–500,000, suitable for high-precision analysis of complex samples.
• Applications: Quantitative proteomics, unknown metabolite identification, metabolic flux analysis.

（2）TOF（Time-of-Flight）

• Principle: Ions pass through an acceleration field and then travel in a vacuum flight tube, with flight time depending on their m/z.
• Features: The longer the flight distance, the higher the resolution; can be combined with MALDI for high-throughput analysis.
• Applications: Rapid metabolite screening, microorganism identification, polymer analysis.

(3) FT-ICR (Fourier Transform Ion Cyclotron Resonance)

• Principle: Ions perform cyclotron motion in a magnetic field, and their frequency signals are analyzed through Fourier transform.
• Features: Extremely high resolution (>1,000,000), but requires high environmental stability, and is relatively costly.
• Applications: Analysis of structurally complex natural products, isotopic fine difference determination.

2. Key Advantages of High-Resolution Mass Spectrometry

High-resolution mass spectrometry demonstrates multiple technical advantages in practical applications:

• It possesses extremely high mass resolution, capable of distinguishing substances with mass differences as small as the third decimal place, such as C₃H₇N (73.057) and C₂H₅O₂ (73.042), which is particularly important for component analysis of complex mixtures.
• HRMS offers excellent mass accuracy, with mass error usually controlled at the ppm level, aiding in the inference of compound molecular formulas and reducing the probability of misjudgment.
• HRMS is typically equipped with MS/MS multi-stage fragmentation analysis capabilities, enabling dual tasks of compound structure analysis and quantitative analysis.

It also has high detection sensitivity and a wide linear dynamic range, capable of detecting trace components in biological samples, making it an important tool for trace analysis of metabolites, proteins, and drugs.

 

3. Advantages of Biotree Biotechnology in High-Resolution Mass Spectrometry Applications

As a leading domestic provider of proteomics and metabolomics services, Biotree Biotechnology relies on high-end mass spectrometry platforms (including Thermo Orbitrap Exploris 480 and Bruker timsTOF Pro 2) to build comprehensive solutions encompassing basic research and translational medicine. Our core advantages include:

• Complete sample pre-processing and standardized mass spectrometry process systems
• Self-developed databases and AI algorithms, enhancing the ability to identify unknown substances
• Providing full-process technical support from project design to result interpretation

High-resolution mass spectrometry is not just a high-precision instrument technology; it is a key support for advancing modern biomedical research from characterization to mechanism exploration. It allows us to identify molecular isomers at the microscopic level, track changes in metabolic pathways, and even achieve early disease diagnosis and precision treatment biomarker screening. Between basic research and clinical translation, HRMS is building a bridge connecting deep data with biological functions. In the future, as AI algorithms and multi-omics integration continue to evolve, high-resolution mass spectrometry will show broader application prospects in fields such as precision medicine, systems biology, and green pharmaceuticals. If you are conducting complex sample analysis, metabolite identification, or protein quantification research, feel free to contact us to experience Biotree Biotechnology's professional services and technical strength in the field of high-resolution mass spectrometry.

Biotree Biotechnology--Characterization of Biological Products, Quality Service Provider for Multi-Omics Biological Mass Spectrometry Detection

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