Mass spectrometry of permethylated folic acid analogs.

Folic acid and several analogs have been characterized as their permethylated derivatives. These derivatives are readily evaporated from the solid probe and provide spectra with abundant molecular ions and characteristic fragment ions. This method has been used to confirm the identities of the two methotrexate metabolites, 7-hydroxymethotrexate and 4-amino-4-deoxy-N(10)-methylpteroic acid, which have been isolated from human urine. The permethylation products of the folate cofactors, dihydrofolic acid, tetrahydrofolic acid and leucovorin, are also characterized.