Optimized method for analyzing volatile compounds in edible yeast using headspace SPME-GC-MS

Volatile compounds produced by edible yeasts play a critical role in food flavor and consumer perception. This study aimed to evaluate how different yeast preparation methods influence the detection of volatile compounds using gas chromatography-mass spectrometry (GC-MS) coupled with headspace solid-phase microextraction (HS-SPME). Saccharomyces cerevisiae was prepared using four methods-broth culture, agar culture, supernatant, and yeast cell pellet-and volatile profiles were compared with non-polar (DB-5), mid-polar (DB-17), and polar (VF-WAX) GC columns. The supernatant consistently exhibited the greatest diversity and abundant volatile compounds, whereas agar cultures and cell pellets led to fewer volatiles. Principal component analysis (PCA) demonstrated distinct clustering of volatile profiles according to the preparation method, with major compounds such as hexanoic acid ethyl ester and phenylethyl alcohol contributing to group separation. Additionally, the effect of salting-out agents (NaCl and H₂NaPO₄) on volatiles extraction efficiency was examined, showing that NaCl led to increased levels of alcohols, while H₂NaPO₄ enhanced acid extraction. These findings underscore the importance of optimizing sample preparation conditions, column polarity, and extraction parameters for accurate and reproducible analysis of yeast-derived volatiles. The results provide practical insights into targeted flavor profiling and the development of yeast-derived flavor applications in the food and fermentation processes.