Mass Spectrometry (MS) has emerged as an indispensable tool for the analysis of biomolecules due to its sensitivity, versatility and ease of applicability to complex samples. Nevertheless, the analysis of free oligosaccharides and protein bound sugars in secretions such as milk poses certain challenges. In this review, the benefits and limitations of different sample preparation approaches for the mass spectrometric analysis of free oligosaccharides and glycoproteins are discussed. Appropriate sample preparation is the first crucial step for successful mass spectrometric analysis. Different MS techniques and instrument combinations already successfully applied to the analysis of milk oligosaccharides are also introduced. Available tandem and MSⁿ applications for the differentiation of structural isomers are described and their limitations discussed. This review is intended to give an overview on the available MS methodology and technology available for analysing various kinds of oligosaccharides in milk.