Presently, only a few clinical laboratories produce their own reference values, while the great majority use reference intervals reported in the literature. An alternative to this unsatisfactory situation is to estimate indirect reference limits by means of mathematical/statistical procedures from patients' results obtained routinely in the laboratory. The procedures of Bhattacharya (A simple method of resolution of a distribution into Gaussian components. Biometrics 1967;23:115–135) Martin et al. (Reference values based on populations accessible to hospitals. In: Gräsbeck R, Alström T, editors. Reference Values in Laboratory Medicine. Chischester: Wiley, 1981:233–262) and Kairisto et al. (Generation of reference values for cardiac enzymes from hospital admission laboratory data. Eur J Clin Chem Clin Biochem 1994;32:789–796) were applied to 14 biochemical quantities. In order to verify these procedures, the indirect reference limits obtained from patients' results were validated by statistical comparison with reference limits estimated from a reference sample according to recommendations of the International Federation of Clinical Chemistry (IFCC). Calculated indirect reference limits for most quantities studied were reliable, but indirect reference limits for bilirubins and potassium ion substance concentrations, alanine aminotransferase, and aspartate aminotransferase catalytic concentrations in serum were not suitable. We conclude that indirect reference limits can be obtained from patients' results by all procedures studied when skewness and kurtosis of mixed distribution are not too large, but other factors also seem to have an influence on the reliability of these procedures.