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LC-MS (Liquid Chromatography-Mass Spectrometry)

A powerful analytical technique that combines the separation capabilities of liquid chromatography with the quantitative and qualitative capabilities of mass spectrometry.

LC MS Sample Prep 1

Sample preparation for LC-MS is the process of extracting, purifying, and concentrating analytes from a sample matrix to be suitable for LC-MS analysis. The primary purpose of LC-MS Sample Preparation is to remove any potential interferences, concentrate the analytes of interest, and make the sample compatible with the LC-MS system to ensure accurate and reliable data.

Workflows involving LC-MS Sample Preparation

LC-MS Sample Prep is commonly used in these workflows:

Proteomics: For protein identification, quantification, and post-translational modification analysis.

Metabolomics: To study small molecules and metabolic pathways.

Pharmaceutical Analysis: For drug discovery, pharmacokinetics, and metabolism studies.

Environmental and Food Analysis: For detecting contaminants or residues.

 

Top methods for LC-MS Sample Preparation

 

Solid Phase Extraction (SPE) 

Solid Phase Extraction (SPE) is a crucial technique in analytical laboratories for sample preparation, especially for chromatographic analyses like LC-MS. This method focuses on isolating analytes from liquid samples using a solid stationary phase, effectively purifying and concentrating them while removing interfering compounds. SPE enhances sample compatibility with chromatographic methods, improves qualitative and quantitative analysis, and extends the lifetime of analytical systems. Notable for its rapid processing and adaptability to automation, SPE is particularly suitable for handling complex matrices such as urine, blood, and food samples, offering superior recovery yields compared to liquid-liquid extraction.

Procedure:

 

Protein Precipitation

Protein Precipitation is a widely used technique aimed at removing proteins from biological samples. This method is essential for preparing samples with high protein content, such as plasma or serum. By precipitating proteins, it simplifies the sample matrix, reducing interference in subsequent LC-MS analysis. The process is favored for its simplicity, speed, and effectiveness in handling large volumes and complex biological matrices. It not only improves the analysis of small molecules but also minimizes the potential for matrix effects that could impact the accuracy and sensitivity of LC-MS analysis.

Procedure:

 

Liquid-Liquid Extraction (LLE) 

Liquid-Liquid Extraction involves separating analytes based on their differential solubilities in two immiscible liquids, typically an aqueous phase and an organic solvent. This method is essential for extracting analytes from complex aqueous matrices, such as biological fluids, and is particularly effective for non-polar or moderately polar compounds. LLE is appreciated for its ability to efficiently separate and concentrate analytes while removing water-soluble interferences, thus enhancing the sensitivity and specificity of LC-MS analysis. The technique is versatile and can be scaled to accommodate various sample volumes.

Procedure:

 

Derivatization for LC-MS Sample Preparation

Derivatization in LC-MS sample preparation is a chemical modification process where reactive groups of analytes are transformed to enhance their detection and quantification. This technique is particularly useful for analytes with low inherent detectability, such as those lacking chromophores or fluorophores. Derivatization improves the physicochemical properties of analytes, such as volatility, stability, and ionization efficiency, making them more amenable to LC-MS analysis. It is crucial for analyzing specific compound classes like steroids, amino acids, and sugars, and can significantly increase sensitivity, selectivity, and overall analytical performance.

 

Why is LC-MS Sample Preparation so difficult:

LC-MS Sample Prep has never been easier

The OT-2 is a bench-top liquid handler designed to be accessible and flexible enough to automate many common applications.

Benefits of Automation over Manual Pipetting for LC-MS Sample Preparation:

Protocol Spotlights

Opentrons helps you automate LC-MS Sample Prep with open-source protocols for the OT-2 and Opentrons Flex

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