Two anal. methods (spectrophotometry, spectrofluorometry) were tested for their applicability for enzymatic detn. of total and free L-carnitine contents in cow, goat and sheep milk samples. The samples were preliminary prepd. by hydrolysis with KOH at 60 degrees C for 1 h, neutralization with HClO(4) and ultrafiltration and then analyzed after enzymatic acetylation of L-carnitine with carnitine acetyltransferase either by colour reaction of the released free coenzyme A with 5,5'-dithio-bis(2-nitro-benzoic) acid (absorption at 412 nm) or by reaction of the coenzyme A with 2-oxoglutarate in presence of its dehydrogenase to hydrogenated nicotin-amide-adenine dinucleotide (absorption at 340 nm). Both methods tested were consistent but the spectrophotometric method was distinguished by higher simplicity. The highest content of total L-carnitine was found in sheep milk (10.8-10.9%) and the lowest in the goat milk (5.7-5.9%). Both the storage of the milk samples at -18 degrees C for 4 weeks and the pasteurization of cow milk at 75 degrees C and 90 degrees C resulted in decrease in the L-carnitine content.