The diagnosis of central nervous system diseases can be a challenge in Veterinary Medicine. When suitable, the exam of the cerebrospinal fluid (CSF), also named liquor, can be of extreme value in the evaluation of those diseases, mainly when evaluated together with patient's history and clinical exam. Even with the great utility of the physicochemical exam of liquor in neurology, few are the studies about their elements' stability under storage. Considering this, the purpose of the present work was to verify the influence of temperature and period of conservation in physicochemical characteristics of liquor of healthy dogs. To perform this, samples of CSF were collected through puncture of cisterna magna of clinically healthy dogs, and then submitted for analysis of density, pH, glycorrhachia, total proteins and activity of the enzymes creatine kinase (CK) and aspartate aminotransferase (AST), after storage in different temperatures (25°C, 4°C, 4°C, 20°C, 70°C) and periods (immediately after collection and after 24 hours, 48 hours, one week and one month). Regarding the results it was possible to verify that, in the samples stored at 25°C, the concentrations of glucose in the CSF, as well as the activity of CK enzyme remain stable for 48 hours and the other parameters appraised for one month. For temperature of cooling, after a month of storage, only glucose concentration suffered statistically significant alteration. For samples kept under freezing temperature of 4°C there was statistically significant alteration in activity of CK after a month of storage, and for the others freezing temperatures (20°C and 70°C), all of the evaluated parameters were stable during all experimental protocol. Considering the results above described, for conservation of the physicochemical parameters of dogs liquor samples, the freezing temperatures 20°C and 70°C are the ideal temperatures of storage.