Objective To evaluate the cold chain in the vaccination points of a health department in the autonomous region of Valencia, including training of the staff responsible for the cold chain. Method We performed a crosssectional study to review all the vaccination points (n=50, with 68 refrigerators) of a health department in the autonomous region of Valencia. A structured questionnaire was used to gather information on the characteristics of the vaccination center, the staff responsible for the cold chain, the infrastructure and control of the elements composing the cold chain, the procedures used in vaccine conservation and handling, and knowledge of the staff. Results The response rate was 100%. Most of the refrigerators were domestic refrigerators without alarm systems to alert staff to technical failure or electrical cuts (76.7%), open door alarm (98.5%), external thermometer (92.6%), automatic defrosting facilities (76.5%), internal temperature sensors or connection to emergency circuits (85.3%). Temperatures were mostly in the correct range (83.9%) and the vaccines were correctly located (88.2%). At least one staff member was responsible for the vaccines in all centers. Food was found in 33.8% of the refrigerators, vaccines were conserved on the vaccination table on the day of vaccination in 32.4%, and the temperature chart was not recorded daily in 75%. Conclusion The appropriate temperature was maintained in five out of every six refrigerators. However, the vaccine infrastructure in primary care centers, as well as training of the staff responsible for the cold chain, should be improved.