Four novel derivatives of azo glycol derivatives (9-12) have been synthesized through multi-step synthetic scheme starting from glycerol. Compounds were also found to be active against gram-positive bacteria (Streptococcus pneumoniae, Staphylococcus epidermidis and Bacillus pumilus) and gram-negative bacteria (Pseudomonus aeruginosa and E.coli) as well as yeast (Candida albicans). In addition, urease inhibition activity of all compounds (9-12) is also reported. Most stable conformation and optimal dihedral angle of glycol moiety is also evaluated by computational method.