CD BioGlyco provides systematic and one-stop services for ergosterol lipid analysis and ensures the continuity and transparency of the experimental process. We will continue to improve our standards to meet customers' needs for sterol lipids analysis.
Due to the increase in the incidence of fungal infections and related antifungal resistance patterns for different species, their accurate and rapid detection is becoming increasingly important in numerous fields. At present, the detection of ergosterol, a fungal biomarker, is considered to be the method of choice. Numerous papers have been published about the use of ergosterol as a general indicator of fungal contamination. Ergosterol is the primary sterol in the cell membranes of filamentous fungi and is either absent or a minor component in the majority of higher plants. It is also present in the membranes of the yeast cell wall. Ergosterol is essential for fungal growth. Ergosterol content has been used to estimate fungal biomass in various environments, e.g. in soil and aquatic systems because a strong correlation exists between ergosterol content and fungal dry mass.
Ergosterol assay is generally considered to be the most promising method for the detection of fungi because of (a) the specificity of ergosterol to fungi, (b) the fact that it indicates live fungal mass (ergosterol becomes oxidized upon cell death), and (c) the relative constancy of the conversion factors compared to other alternative methods.
Fig.1 HPLC analysis of total cellular ergosterol. (South, et al.,2013)
Ergosterol is the major sterol constituent of most fungi. Since it is present in negligible amounts in higher plants, it can be used as a chemical marker for the presence of fungal contamination. CD BioGlyco provides various ergosterol lipid analysis services. Our technology for ergosterol lipid analysis includes but is not limited to:
Sterols are extracted from yeast using potassium hydroxide in ethanol. After extraction, nonpolar lipids are separated using N-hexane two times. Nonpolar sterols are crystallized after evaporation of the N-hexane and redissolved in methanol. Samples are then analyzed and separated using a C-18 column. A standard curve is determined using known concentrations of purified ergosterol.
This method enables accurate, rapid, and efficient determination of ergosterol content. Such a method allows for minimizing the potential degradation of ergosterol because of light exposure, with a subsequent decrease in false-negative results.
Samples are saponified in methanolic KOH. Ergosterol is extracted by hexane extraction and subsequently silylated by N-trimethylsilylimidazole / trimethylchlorosilane reagent at room temperature. The method can be easily applied to handling a large number of samples.
The sample is evaporated on a vacuum evaporator and suspended in methanol. Compounds are separated using an HP-5 capillary column with a non-polar stationary phase. The carrier gas used is helium.
We constantly update our technology platform to provide customers with comprehensive and high-quality lipidomics analysis services. CD BioGlyco is committed to raising the standard to meet customers' ergosterol lipid analysis needs. If you are interested in our services, please feel free to contact us. We are looking forward to being your indispensable assistant.
Reference: