Guanidino compounds in cerebrospinal fluid (CSF) of nonneurological and epileptic patients were investigated. After separating using high performance liquid chromatography, and reacting with phenanthrenequinone, the compounds were analyzed fluorometrically. Arginine (Arg), creatinine (CRN) and homoarginine (HArg) were found in all subjects. Trace amounts of guanidinosuccinic acid (GSA) and guanidinoacetic acid (GAA) were detected in some subjects regardless of disease state. Positive correlations were noted in females between Arg and CRN levels and age. Serum Arg and HArg levels were related to in urine concentrations, but there was no association between CSF guanidino compound levels and serum levels. Guanidinoethanesulfonic acid (GES) was detected in the CSF of some epileptic patients. This was confirmed by high voltage paper electrophoresis. This finding suggests that GES may relate to epileptic seizure mechanism.