A variety of compounds which bind to Fe have substantial reductions in absorption when co-administered with Fe compounds. The binding of both Fe2+ and Fe3+ ions to pteroylmonoglutamic acid and the pteroylmonoglutamate dianion was examined in vitro. In dimethylsulphoxide (DMSO) alone, pteroylmonoglutamate formed a 2:1 (pteroylmonoglutamate: Fe3+ ion) complex. However, in DMSO–aqueous Bis-Tris buffer (4:1, v/v; pH 6.0) no evidence of complex formation could be seen. Likewise spectroscopic evidence was obtained for complex formation with Fe2+ ion and pteroylmonoglutamate in DMSO alone but not in the aqueous DMSO buffer. In vivo studies examined the effect of FeSO4 on pteroylmonoglutamic acid absorption in an isolated perfused rat jejunal model of nutrient absorption. The dose of pteroylmonoglutamic acid approximated a human dose of 1 mg for the rat, while the FeSO4 doses were chosen to represent 6.4 mg, 64 mg and 300 mg human doses. There was no significant effect of FeSO4 on pteroylmonoglutamic acid absorption or instability of pteroylmonoglutamic acid in vivo in the presence of FeSO4 in the rat. Although 2:1 binding of pteroylmonoglutamic acid to Fe ions could be demonstrated in DMSO alone, no binding could be demonstrated in DMSO–Bis-Tris buffer (4:1, v/v; pH 6.0). It is unlikely that there will be a significant reduction in pteroylmonoglutamic acid absorption during concurrent ingestion of Fe preparations.