Two-Dimensional GC–ToFMS Analysis of Volatile Organic Compounds in Fermented Camel Milk (Shubat)

Shubat, a traditional fermented camel milk from Kazakhstan, is renowned for its unique flavor and nutritional properties, though its volatile compound profile remains poorly characterized. In this study, headspace solid-phase microextraction coupled with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (HS-SPME-GC×GC-ToFMS) was employed to qualitatively identify and semi-quantitatively analyze volatile metabolites in seven Shubat samples collected from four regions of Kazakhstan. Of the 372 volatile organic compounds initially detected, 202 were retained after screening, predominantly comprising esters, acids, alcohols, ketones, and aldehydes. Esters, acids, and alcohol were found to be the most abundant categories. Diversity analyses (α and β) revealed substantial variation across regions, likely influenced by Shubat's rich and region-specific microbiome. An UpSet analysis demonstrated that 75 volatile compounds were shared among all samples, accounting for over 87% of the total volatile content, indicating a chemically stable core. These findings underscore the chemical complexity of Shubat and provide novel insights into its metabolite composition, thereby establishing a foundation for future sensory, microbial, and quality-related research.