As determined by a colorimetric assay measuring parent compounds plus ether-extractable nitroso-containing metabolites, N,N'-bis(2-chloroethyl)-N-nitrosourea (BCNU) disappeared more rapidly than N-(2-chloroethyl)-N'cyclohexyl-N-nitrosourea (CCNU) and N-(2-chloroethyl)-N'-(4-transmethylcyclohexyl)-N-nitrosourea (MeCCNU) and their products from the tissues of mice injected IV. Assay of selected samples by high-pressure liquid chromatography revealed that the colorimetric assay for BCNU was specific in that the two assays gave equivalent results. Following IV-injections of CCNU and MeCCNU, however, levels of the parent compounds decreased much more rapidly than the total, color-producing material. Of seven tissues, the largest Cxt values for BCNU, as determined by the colorimetric assay, were noted for blood (442 microgram-min/ml) and large intestine (285 microgram-min/g). Liver (29 microgram-min/g) had the lowest Cxt value, reflecting rapid metabolism of the compound by this organ. Color-producing material related to CCNU disappeared from the solid tissues of mice in a manner generally parallel to that for blood. Of the Cxt values for this compound and its products, those for lung (1753 microgram-equivalents-min/g), kidney (1633 microgram-equivalents-min/g), and small intestine (1557 microgram-equivalents-min/g) were highest. Consistent with its slower rate of metabolism, MeCCNU and its color-producing metabolites remained in most tissues of mice for 12 h following injection. Except for brain (1434 microgram-equivalents-min/g), Cxt values for this nitrosourea and its metabolites in tissues were higher than those of blood (5485 microgram-equivalents-min/ml), with kidney (15,324 micrograms-equivalents-min/g), liver (12,921 microgram-equivalents-min/g), and large intestine (11,501 microgram-equivalents-min/g) being highest. For each nitrosourea, a fair correlation was observed between the Cxt values for tissues and the toxic and/or antitumor effects at those sites.
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