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What is MRM

Multiple Reaction Monitoring (MRM) is a quantitative mass spectrometry technique that enables highly sensitive and selective detection of target molecules by monitoring specific precursor-product ion transitions.

 

Principle of MRM

MRM operates on a triple quadrupole mass spectrometer. In this mode, the first quadrupole (Q1) selectively filters a precursor ion of interest. The ion then undergoes fragmentation via collision-induced dissociation (CID) in the second quadrupole (q2). Finally, the third quadrupole (Q3) isolates and detects a specific product ion, ensuring high selectivity and sensitivity.

 

Advantages of MRM

1. High Selectivity

By monitoring predefined precursor-product ion pairs, MRM effectively eliminates interference from non-target compounds, improving specificity.

 

2. High Sensitivity

Selective monitoring of specific ion transitions reduces background noise, enhancing signal detection and overall sensitivity.

 

3. Accurate Quantification

MRM employs internal standards for quantification. By introducing an internal standard, variations in sample preparation and instrumental response are corrected, ensuring reliable and reproducible quantification.

 

Applications of MRM in Biopharmaceutical Research

1. Drug Metabolism Studies

MRM facilitates the quantitative analysis of drug metabolites, providing insights into metabolic pathways and metabolite formation in vivo.

 

2. Pharmacokinetic Analysis

MRM enables precise quantification of drug absorption, distribution, metabolism, and excretion (ADME), facilitating pharmacokinetic assessments.

 

3. Protein Quantification

MRM is widely applied in targeted protein quantification, including the measurement of drug-target protein levels to elucidate drug-target interactions.

 

Experimental Workflow of MRM

1. Selection of Precursor-Product Ion Pairs

Suitable precursor-product ion transitions are identified based on the molecular structure, fragmentation pattern, and mass-to-charge ratio of the target compound.

 

2. Optimization of Collision Energy

The collision energy is fine-tuned to maximize the generation of product ions, ensuring optimal signal intensity.

 

3. Determination of Quantifier ions

The most suitable precursor-product ion transitions are chosen for precise quantification.

 

4. Calibration Curve Construction

A series of standard solutions with known concentrations are prepared to construct a calibration curve, enabling accurate quantification of the target molecule.

 

5. Sample Preparation and Analysis

Appropriate sample preparation techniques, such as extraction and purification, are applied before mass spectrometric analysis to ensure data accuracy and reproducibility.

 

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