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It is a cytotoxic nitrogen mustard derivative widely used in cancer chemotherapy. It cross-links DNA, causes strand breakage, and induces mutations. Its clinical activity is associated with a decrease in aldehyde dehydrogenase 1 (ALDH1) activity. This substance is listed as a known human carcinogen.

Cyclophosphamide-d₄ contains four deuterium atoms. It is intended for use as an internal standard for the quantification of cyclophosphamide by GC- or LC-MS. Cyclophosphamide is a nitrogen mustard alkylating agent used in the treatment of cancers and autoimmune disorders. In cells with low levels of aldehyde dehydrogenase, cyclophosphamide acts as a prodrug and is metabolized to the active compound phosphoramide mustard, which crosslinks with DNA and causes cell death.