Lactose is a disaccharide composed of galactose and glucose and has the molecular formula C₁₂H₂₂O₁₁. Lactose makes up around 2–8% of milk (by mass). The name comes from lac (gen. lactis), the Latin word for milk, plus the suffix -ose used to name sugars. The compound is a white, water-soluble, non-hygroscopic solid with a mildly sweet taste. It is used in the food industry.

Lactose is a disaccharide composed of galactose and glucose, which form a β-1→4 glycosidic linkage. Its systematic name is β-D-galactopyranosyl-(1→4)-D-glucose. The glucose can be in either the α-pyranose form or the β-pyranose form, whereas the galactose can have only the β-pyranose form: hence α-lactose and β-lactose refer to the anomeric form of the glucopyranose ring alone. Detection reactions for lactose are the Wöhlk and Fearon tests. They can be used to detect the different lactose content of dairy products such as whole milk, lactose free milk, yogurt, buttermilk, coffee creamer, sour cream, kefir, etc.

Lactose is hydrolysed to glucose and galactose, isomerised in alkaline solution to lactulose, and catalytically hydrogenated to the corresponding polyhydric alcohol, lactitol. Lactulose is a commercial product, used for treatment of constipation.

Lactose appears in the milk of many different mammal species, though not all. Rhesus macaque monkeys have a concentration of lactose of 8%. Many mammals, like bears, produce milk with no lactose. Bovine milk has a concentration of around 4.8%.

Whey or milk plasma is the liquid remaining after milk is curdled and strained, for example in the production of cheese. Whey is made up of 6.5% solids, of which 4.8% is lactose, which is purified by crystallisation. Industrially, lactose is produced from whey permeate – whey filtrated for all major proteins. The protein fraction is used in infant nutrition and sports nutrition while the permeate can be evaporated to 60–65% solids and crystallized while cooling. Lactose can also be isolated by dilution of whey with ethanol.

Infant mammals nurse on their mothers to drink milk, which is rich in lactose. The intestinal villi secrete the enzyme lactase (β-D-galactosidase) to digest it. This enzyme cleaves the lactose molecule into its two subunits, the simple sugars glucose and galactose, which can be absorbed. Since lactose occurs mostly in milk, in most mammals, the production of lactase gradually decreases with maturity due to weaning; the removal of lactose from the diet removes the metabolic pressure to continue to produce lactase for its digestion.

Many people with ancestry in Europe, West Asia, South Asia, the Sahel belt in West Africa, East Africa and a few other parts of Central Africa maintain lactase production into adulthood due to selection for genes that continue lactase production. In many of these areas, milk from mammals such as cattle, goats, and sheep is used as a large source of food. It was in these regions that genes for lifelong lactase production first evolved. The genes of adult lactose tolerance have evolved independently in various ethnic groups.^{[page needed]} By descent, more than 70% of western Europeans can digest lactose as adults, compared with less than 30% of people from areas of Africa, eastern and south-eastern Asia and Oceania. In people who are lactose intolerant, lactose is not broken down and provides food for gas-producing gut flora, which can lead to diarrhea, bloating, flatulence, and other gastrointestinal symptoms.

The sweetness of lactose is 0.2 to 0.4, relative to 1.0 for sucrose. For comparison, the sweetness of glucose is 0.6 to 0.7, of fructose is 1.3, of galactose is 0.5 to 0.7, of maltose is 0.4 to 0.5, of sorbose is 0.4, and of xylose is 0.6 to 0.7.