Gas chromatography analysis of major free mono- and disaccharides in milk: Method assessment, validation, and...

Saccharides are functional constituents of milk. Although d-lactose represents almost the totality of the saccharides in the milk, minor species, like d-glucose, d-galactose, myo-inositol and, as a result of severe thermal treatments, monosaccharides like d-tagatose, are also detectable. Although chromatography has been the main analytical approach used for accomplishing this task, quite surprisingly a validated gas chromatographic method aimed at the simultaneous determination of these compounds is still needed. Hence, our contribution is devoted to fill this gap. After the optimization of clean-up and derivatization (conversion of saccharides in their trimethyl silyl ethers) steps, the adoption of a highly cross-linked silphenylene stationary phase permitted to obtain high resolution and a fast chromatographic run. Validation was accomplished in terms of limit of detection, limit of quantification, linearity, precision, and trueness. The accuracy of the method was successfully tested on a number of partially skimmed milk samples. Excellent limits of detection for all analytes make this method eligible, also with respect to a gas chromatographic/mass spectrometry approach, for routine analysis and quality control in the dairy industries.