Abstract Background This study aimed to define RIs for red blood cell series and total leukocytes in the complete blood count, in adult patients. Methods We used a sample of 822 patients aged 18 to 60 years, both female and male. The study focused on the red blood cell series (RBC Count, hemoglobin, Hematocrit, MCV, MCH, MCHC and RDW) and total leukocytes. RIs and respective 90% confidence intervals were estimated using the refineR method and a multi-criteria method combining the Expectation Maximization algorithm, mode, antimode analysis, an adapted Horn algorithm (5 cycles of Tukey's test with intermediate trimming and 2 cycles of Tukey's test with Box-Cox transformed data if applicable after analyzing Lambda, kurtosis, and skewness coefficients) and distribution truncation based on clusters and antimode. The RIs estimates using refineR and multi-criteria methods were compared to the manufacturer's specifications (Mindray). Flagging Rates were used to validate the method that estimates the most representative RI for the population served by the laboratory. Equivalence Interval and Bias Ratio were utilized to assess the practical relevance of sex-based partitioning. Results The method based on the multicriteria approach demonstrated the best performance, with flagging rates showing results lower than 4.7% and higher than 0.9% after the truncation of the results distribution. In general, the refineR method generated very narrow RIs, while the manufacturer's specifications resulted in very wide or shifted RIs, which do not adequately represent the data distribution profile. It was identified the need to partition by sex for the parameters Red Blood Cell Count, Hemoglobin, and Hematocrit, which is in agreement with the manufacturer's specifications. Conclusions The estimated RIs better represent the laboratory-served population, highlighting the importance of adhering to ISO 15189 normative requirements. This involves verifying and/or determining RIs to provide representative information of the served population, thereby aiding in medical decision-making.