Calibration problems connected with the determination of C20-C44 petroleum hydrocarbons by gas chromatography – flame ionization detection (GC-FID) and IR spectrometric methods were studied in this paper. It has been found that the mass responses and mass response factors of FID determined by capillary GC-FID as well as by IR spectrometry, at wave number 2959 and 2924 cm-1, of high boiling petroleum distillation fractions (in the range C20-C46) are not constant. Moreover, it was found that mass response factors for differentpetroleum oils that have the same boiling temperature interval might differ up to 1.5 fold both for FID and IRD methods, dependingon their hydrocarbon group composition. However, calibration of the IR spectrometric method using the empirical equations proposed in DIN 38 409, Part 18, norm (for the determination of total petroleumhydrocarbons in waters polluted with high boiled petroleum fractions)gave results which overlapped within ±10% of the expected values.