BackgroundHematological reference values are important for the clinical decisions in laboratory diagnosis and monitoring of patients. The correct interpretation of laboratory results depends entirely on the reference intervals that have been established for the locality. But, in sub-Saharan African countries particularly in Ethiopia, locally derived reference intervals were not established and they are forced to use intervals established from western population. Thus this study aimed to establish locally derived hematological reference values that could be used in Northwest Ethiopia.MethodsA cross sectional study was conducted from April to May 2014 with 120 male and 120 female apparently health adult blood donors at Gondar University Hospital. A structured pretested questionnaire was used for socio demographic and clinical data collection. About 4 ml of blood was collected with EDTA test tube and analyzed using Cell-Dyn 1800 to enumerate the hematological parameters. The data were collected and entered into SPSS version 20 for analysis. Mann–Whitney U test was used to determine reference intervals and Harris and Boyd test was used to determine the reference intervals that need partition. The 95th percentile of measurements was taken as a reference interval.ResultsMedian and 95th percentile of WBC for general population were lower than Caucasian population, Addis Ababa, Burkina Faso and Kenya of similar studies. The RBC, Hgb and PCV lower 95% limit values of both sex were lower than studies in Addis Ababa, Kenya, Burkina Faso and text book. While PCV upper limit values higher than the above countries. MCV values of the current study were higher than those countries while MCHC values were lower. Similarly, the absolute values of neutrophils in the current study were lower than Caucasian and Afro Caribbean but higher than African countries and Jamaica but lymphocyte count was higher.ConclusionsThe hematological reference intervals established in this study was different from those reported in other part of Ethiopia or African countries as well as Caucasian population. The RBC, PCV, Hgb and MCHC reference intervals were different in gender. Thus, using of locally determined reference range is advisable.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-016-2288-8) contains supplementary material, which is available to authorized users.