Vancomycin belongs to the vancomycin-ristocetin family of glycopeptides, and is a subclass of linear sugar containing peptides composed of seven amino acids. Its stereochemical configuration forms the basis of a particular mode of action, though its complexation with the D-alanyl-D-alanine terminus of peptidoglycon monomer. The glycosylated hexapeptide chain consists of chloro-β-hydroxytyrosines, p-hydroxyphenylglycines, N-methylleucine and aspartic acid forms a rigid molecular frame work and gives the difficulty in the analysis. Vancomycin in the serum samples is usually estimated by liquid chromatography and the bacterial sensitivity was genereally tested by the microbiological assay. The present review deals with the qualitative, quantitative, microbiological and immunological assays and the comparison of the quantitative methods. Clinical implications of vancomycin have also been cited in the review.