Increasing sales of organic milk mean intensified tests for authenticity are required. In addition to comprehensive documentation, analytical methods to identify organic milk, and thus to differentiate it from conventional milk, are needed for consumer protection. Because the composition of milk is fundamentally dependent on the feeding of the cows, thirty-five samples from both production systems in Germany, including farm and retail milk, were collected within 12 months, to reflect seasonal variation, and appropriate properties were analysed. Fatty acid analysis enabled organic and conventional milk to be completely distinguished, because of the higher alpha-linolenic acid (C18:3omega3) and eicosapentaenoic acid (C20:5omega3) content of the former. Organic milk fat contained at least 0.56% C18:3omega3 whereas the maximum in conventional milk was 0.53%. Because of the parallel seasonal course of the C18:3omega3 content of organic and conventional retail samples, however, time-resolved comparison at the five sampling dates resulted in a clearer difference of 0.34+/-0.06% on average. Analysis of stable carbon isotopes (delta13C) also enabled complete distinction of both types of milk; this can be explained by the different amounts of maize in the feed. For conventional milk fat delta13C values were -26.6 per thousand or higher whereas for organic milk fat values were always lower, with a maximum of -28.0 per thousand. The time-resolved average difference was 4.5+/-1.0 per thousand. A strong negative correlation (r=-0.92) was found between C18:3omega3 and delta13C. Analysis of a larger number of samples is required to check the preliminary variation ranges obtained in this pilot study and, probably, to adjust the limits. Stable isotopes of nitrogen (delta15N) or sulfur (delta34S) did not enable assignment of the origin of the milk; in cases of ambiguity, however, some trends observed might be useful in combination with other properties.