The aim of this study was to evaluate the levels of plasma homocysteine (Hcy), vitamin B12, and folic acid in patients with pseudoexfoliation glaucoma (PEXG), pseudoexfoliation syndrome (PEXS), PEXS plus normotensive glaucoma (NTG). In total, 24 patients with PEXG, 35 patients with PEXS, 18 patients with PEXS plus NTG, and 35 control subjects were enrolled into study. Their Hcy levels were measured by high performance liquid chromatography (HPLC); the levels of vitamin B12 and folic acid were measured by a competitive electrochemiluminescence immunoassay. Higher plasma Hcy levels and lower folic acid and vitamin B12 levels were found in all 3 patients' groups compared with the control group (all P<0.001, expect for folic acid in the PEXG group, P=0.03). Although plasma Hcy levels in the PEXG and PEXS groups were similar, the PEXS plus NTG group had significantly higher Hcy levels compared with these groups (P=0.019 and P=0.032, respectively). Our study showed that there was an association between hyperhomocysteinemia and PEXS either with or without glaucoma. The patients with PEXS plus NTG had higher plasma Hcy levels than the patients with PEXS or PEXG and the healthy controls. The treatment of hyperhomocysteinemia by taking low-cost vitamin B12 and folic acid preparations may prevent additional vascular problems.