As average lifespan increases, so too does the prevalence of neurodegenerative disorders such as Normal Pressure Hydrocephalus (NPH). NPH can result in dementia symptoms, gait disturbance and incontinence, however drainage of CSF through a lumbar peritoneal shunt can relieve these symptoms for some patients. Why this condition occurs, and how treatment brings about relief is still not clearly understood despite decades of research.Folate has been shown to be correlated with several neurological disorders, and our previous work showed that there is evidence for changes in folate cycle components in the CSF of NPH patients as a result of drainage (Miyan, et al., 2013). The aim of this work is to further investigate the possible role of folate metabolism in NPH patients (n=5) relative to Controls (n=15) at the beginning of the drainage procedure.CSF was collected from patients in accordance with the method published previously by Woodruff et al., (2013) and analysed in triplicate for 5-methyltetrahydrofolate (5MTHF), B6, B12, Homocysteine, Cysteinebetasynthase (CBS) and Formyltetrahydrofolatedehydrogenase (FDH) using a standard dot blot technique.Both 5-MTHF and CBS levels were found to be reduced in NPH patients compared to controls, whereas B12 and B6 levels did not differ. Homocysteine was found to be increased along with FDH, which was found to be increased by up to tenfold in NPH patients when compared to controls.Reduced 5-MTHF levels have been shown to result in hyperhomocysteinaemia, and Homocysteine has previously been shown to have a negative correlation with conditions including dementia and stroke (Smith et al., 2010), CBS activity is closely related to Homocysteine, and this part of the folate pathway has been linked to neurotransmitter production eg. serotonin, and dopamine (Bottiglieri, et al., 2000). The results of this study confirm the CSF of NPH patients shows differences to that of normal patients, related to detrimental disruptions the folate pathway. These findings confirm the possibility of a link between folate metabolism and NPH. The role of FDH in NPH requires further investigation, if only as a possible marker for diagnosis of this condition.