The use of solid phase microextraction for metabolomic analysis of non-small cell lung carcinoma cell line (A549) after administration of combretastatin A4

Abstract

Use of solid phase microextraction (SPME) for cell culture metabolomic analysis allows for the attainment of more sophisticated data from in vitro cell cultures. Moreover, considering that SPME allows the implementation of multiple extractions from the same sample due to its non/low-depletive nature, time course studies using the same set of samples are thus facilitated via this method. Such an approach results in a reduction in the number of samples needed for analysis thus eliminates inter-batch variability related to biological variation occurring during cell culturing. The current work aims to demonstrate the capability of SPME for measurements of combretastatin A4 (CA4) effectiveness on non-small cell cancer cell line. A cultivation protocol was established in the 96-well plate, and a fiber format of SPME was selected for metabolite extraction. The extracellular metabolic pattern of cells was changed after administration of the tested drug. This suggests pharmacological activity of the administered compound towards the studied cell line model. Results support that the use of direct immersion SPME for analysis of cell cultures does not affect cells growth or contaminate sample. Consequently, SPME allows the attainment of accurate information regarding drug uptake, metabolism, and metabolomic changes in the studied cells induced by exposure to the drug simultaneously in a single experiment.

Recurrent Topics in Mass Spectrometry-Based Metabolomics and Lipidomics—Standardization, Coverage, and Throughput

“…While multianalyte quantification remains a challenge, low invasiveness of SPME and the nonexhaustive nature of extraction, together with recently developed extractive phases, make the technique particularly attractive for time-resolved or spatially resolved metabolomics fingerprinting . For example, a high-throughput time-course metabolomic analysis was achieved through multiple extraction of 96-well-plate cell cultures . Direct immersion (DI) in vivo sampling enabled time-resolved metabolic fingerprinting of animal brains , and a method for the analysis of small molecules from semi-solid tissue relying on DI-SPME and desorption electrospray ionization, (DESI)-MS, has been proposed, promising space-resolved analysis of tissues .…”

Toxicometabolomics: Small Molecules to Answer Big Toxicological Questions

“…The extraction method must be highly efficient and nonselective, and its choice presents a great challenge due to the heterogeneity of the metabolome. Liquid–liquid extraction is one of the most applied extraction methods [ 76 , 77 , 78 ], although several other methods can be used (e.g., supercritical fluid extraction [ 79 ], solid-phase extraction [ 80 ], or solid-phase microextraction [ 81 ]). Depending on the planned analysis, a monophasic (such as water/methanol, water/acetonitrile, 100% methanol) or biphasic (water and methanol, often associated with a nonpolar solvent such as chloroform or dichloromethane) solvent solution, in varied proportions and temperatures, can be used for extraction of a large panel of metabolites [ 29 , 73 , 77 , 82 , 83 ].…”

One extraction tool for in vitro-in vivo extrapolation? SPME-based metabolomics of in vitro 2D, 3D, and in vivo mouse melanoma models

“…Coupling of SPME with high-resolution mass spectrometry (HRMS), e.g., orbitrap mass spectrometer or time-of-flight, enables detection of metabolites with high mass accuracy and thus precise identification of these low molecular weight compounds. SPME has also been shown to be suitable for extraction of metabolites excreted by cell cultures [ 10 , 11 ].…”