The K-LOLAC test kit offers a rapid, novel, sequential measurement of free-glucose and lactose in conventional, low-lactose and lactose-free dairy products. This sequential assay format reduces the manual input required by an analyst when compared to traditional lactose assay formats and therefore improves both accuracy and efficiency. When used in combination with the Megazyme Creep Calculator provided, the β-galactosidase employed in this kit allows for the selective measurement of lactose in the presence of galacto-oligosaccharides (GOS) which are commonly found in lactose-free dairy products. This constitutes a significant improvement over existing commercially available lactose assay kits which typically overestimate lactose content in lactose-free samples due to the unselective hydrolysis of GOS by β-galactosidase. Lastly, the sensitivity of the K-LOLAC assay kit has been doubled through the use of a cascade biochemical pathway, helping to significantly reduce the LOD and LOQ for the assay.