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Galantamine (GAL), donepezil (DON) and rivastigmine (RIV) are cholinesterase inhibitors administered to patients who suffer from Alzheimer's disease (AD). We have currently developed and validated an HPLC-DAD method for the determination of GAL, DON, RIV in cerebrospinal fluid (CSF), blood serum and urine. The retention times of the drugs were 1.5, 2.2 and 3.1 min, respectively and the total time of analysis was 5 min. Validation was performed in terms of linearity, selectivity, accuracy, precision and stability. Following validation, the method was applied to clinical CSF, blood serum and urine samples. The currently developed method is a reliable tool for monitoring CSF, serum and urine galantamine, donepezil and rivastigmine levels in patients under treatment with these drugs.