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Analysis of Metabolites by Liquid Chromatography-Mass Spectrometry (LC-MS) (CAT#: STEM-CT-1638-CJ)

Introduction

In biochemistry, a metabolite is an intermediate or end product of metabolism. The term is usually used for small molecules. Metabolites have various functions, including fuel, structure, signaling, stimulatory and inhibitory effects on enzymes, catalytic activity of their own (usually as a cofactor to an enzyme), defense, and interactions with other organisms (e.g. pigments, odorants, and pheromones).




Principle

Liquid chromatography (LC) is a technique widely used to separate compounds from a sample prior to analysis and is frequently coupled to mass spectrometry (MS). With LC, separation of the sample components is based on the interactions of the compounds with the mobile and stationary phases, and the degree of compound separation is related to each compound's affinity for the mobile phase. Following chromatographic separations, compounds elute off of the column, desolvated into the gas phase and ionized at an ionization source, and are then introduced into the mass spectrometer for mass analysis.

Applications

Biochemistry; Biomedical

Procedure

1. Sample Collection
2. Sample Preparation
3. Standardization and Calibration
4. Chromatographic Separation and Analysis

Materials

• Sample: Blood; Urine; Plasma; Crops; Livestock tissues; Milk; Eggs; Soil; Water; Sediment; Nucleotides; Peptides; Steroids; Hormones; Dyes; Fatty acids; Alcohols
• Equipment: Liquid-phase separation system; Mass spectrometer; Liquid chromatograph mass spectrometers
• LC-MS grade water; Acetonitrile
• Ion pairing agents: Trifluoroacetic acid (TFA); Formic acid (FA); Heptafluorobuteric acid (HFBA)

Notes

1. LC-MS is a dominant analytical technique that has a very high accuracy, sensitivity, and specificity.
2. Although known for its key features in providing rapid analysis of complex mixtures and quality performance, LC-MS is an expensive instrument.