Metabolomics is the comprehensive assessment of endogenous metabolites of a biological system. "Oncometabolomics" is a rapidly emerging field with potential for developing specific biomarkers for early detection, diagnosis, and disease prognosis. Given the power of this technology, the availability of standardized sample preparation methods for immortalized human cancer cell lines is critical toward augmenting research in this direction. Using MCF-7 cells as a model system, we describe an approach for intracellular metabolite extraction from cell cultures for reproducible and comprehensive metabolite extraction. The samples, when injected onto a reverse-phase 50 × 2.1 mm Acquity 1.7-μm C18 column, using an ultra performance liquid chromatography system (UPLC) coupled with electrospray ionization-quadrupole-time-of-flight-mass spectrometry (ESI-Q-TOF-MS) in positive and negative modes, yielded a data matrix with a total of 2600 features. This method, when compared with a water-extraction procedure described earlier, was found to yield significantly higher coverage and detection of molecular features. Finally, we successfully tested the performance of this method for an array of human cancer cell lines used widely in the cancer research field.
The article is published online in The Journal of Biomolecular Techniques and is free to access.
Small Molecule Metabolite Extraction Strategy for Improving LC/MS Detection of Cancer Cell Metabolome
News Apr 06, 2011
In the womb, a human fetus has limited contact with pathogens. However, once born, babies face a myriad of germs completely new to their bodies. Their immune system must rapidly develop to ensure early protection from infection. But what is exactly the dynamic of the immune system development in the first days of life?READ MORE
In humans, vitamin D is formed in the skin following its exposure to sunlight. In comparison to the body's own formation of vitamin D, dietary consumption generally makes up only a relatively small proportion of the vitamin D supply to the body. While an overdose resulting from the body's own production is not possible, it certainly can result from the consumption of highdoses of vitamin D - such as via certain food supplements.READ MORE