同位素分离通过将某种化学元素的其它类型的同位素去除而达到浓缩某种特殊的同位素的目的。例如，通过同位素分离可以将天然铀分离成浓缩铀和贫铀，这是为核电站以及铀核武器制造铀燃料的关键技术。钚核武器所使用的钚在反应堆中制成，同样需要制备某种特别的同位素。同位素分离的理论由查尔斯·汤斯首先提出。化学元素的提纯可以通过化学过程，但是由于相同元素的同位素拥有几乎完全相同的化学性质，使得化学方法几乎无法分离同位素，除非是分离氘。

Isotope separation is the process of concentrating specific isotopes of a chemical element by removing other isotopes. The use of the nuclides produced is various. The largest variety is used in research (e.g. in chemistry where atoms of "marker" nuclide are used to figure out reaction mechanisms). By tonnage, separating natural uranium into enriched uranium and depleted uranium is the largest application. In the following text, mainly the uranium enrichment is considered. This process is a crucial one in the manufacture of uranium fuel for nuclear power stations, and is also required for the creation of uranium based nuclear weapons. Plutonium-based weapons use plutonium produced in a nuclear reactor, which must be operated in such a way as to produce plutonium already of suitable isotopic mix or grade. While different chemical elements can be purified through chemical processes, isotopes of the same element have nearly identical chemical properties, which makes this type of separation impractical, except for separation of deuterium.