Rapid, sensitive and facile measurement of intra- and extracellular lactate and pyruvate in cells by LC-MS/MS
Year: 2025
Type: article
Abstract: Lactate and pyruvate are essential metabolites of cellular function. They are involved in mitochondrial function and cellular energetics. Their quantification in extra- and intracellular matrices is crucial to understand mitochondrial alterations that could be associated with certain pathologies, such as cancer. In this study, we present an optimized protocol for the simultaneous quantification of lactate and pyruvate in both intra- and extra-cellular compartments by LC-MS/MS. Lactate and pyruvate were separated on a HILIC stationary phase with isocratic elution in less than 4 minutes and using a mobile phase of acetonitrile/H 2 O (60/40). Recoveries were in the range of 84.15% - 102.85% and relative standard deviation of intra-day an inter-day precision was lower than 6.53%. The method was validated according to the accuracy profile method. The lower limit of quantification (LLOQ) was 1.78 μM for lactate and 0.18 μM for pyruvate. This method was applied to extract and quantify the intracellular and extracellular lactate and pyruvate from two respiratory cell lines, BEAS-2B and A549, cultivated in a 24-well plate. The developed technique is fast, sensitive, specific, reproducible and could be adaptable to various cellular models. • The method involves fast sample preparation and analysis • A high level of sensitivity, accuracy and precision was observed in the method • This method allows the measurement of intra- and extracellular lactate and pyruvate • This method is adaptable to many cellular models and other complex matrices • The method is optimized for a 24-well plate, allowing the replication of experimental conditions
Source: Talanta
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,610
Funder: Université de Rouen
OpenAlex ID: W4415929584