Development of a new isotopically labeled internal standard for ergosterol measurement by GC/MS.

Joanne Sordillo, Donato Vespa, Linda Haggerty, Frederick Youngs, Diane Gold, Donald Milton
Author Information
  1. Joanne Sordillo: Channing Laboratory, Boston, MA, USA.

Abstract

Environmental exposure to fungi has been associated with a variety of adverse health effects. Ergosterol, a marker of total fungal biomass, can be used to quantify exposure to fungi. Unfortunately, environmental ergosterol measurement using published GC/MS/MS methods is prone to bias introduced by sample matrix effects, resulting in potential measurement inaccuracy. We developed an isotopically labeled internal standard ((13)C ergosterol) for ergosterol quantification by GC/MS/MS, to eliminate bias due to sample matrix effects and selective losses during preparation. To produce (13)C ergosterol, we grew Saccharomyces Cerevisiae on (13)C D-glucose under aerobic conditions at room temperature. The (13)C labeled ergosterol comprised 97.1% of the ergosterol in the dry yeast preparation. Ergosterol spike-recovery from house dust samples averaged 99.3% using the isotopically labeled yeast preparation as the internal standard (I.S.). By contrast, spike-recovery averaged 42.4% when 7-dehydrocholesterol (7-DHC) was the internal standard. Analysis of indoor house dust samples from a large epidemiologic study also showed the systematic underestimation of ergosterol when analyzed using 7-DHC as compared with using the yeast preparation containing the isotopically labeled authentic compound, (13)C-ergosterol.

MeSH Term

Carbon Isotopes
Dehydrocholesterols
Ergosterol
Gas Chromatography-Mass Spectrometry
Glucose
Isotope Labeling
Reference Standards
Saccharomyces cerevisiae

Chemicals

Carbon Isotopes
Dehydrocholesterols
7-dehydrocholesterol
Glucose
Ergosterol

Word Cloud

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