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This paper describes the most significant achievements related to the creation and development of separation methods. Tswiett Nerst’s achievements and the role of such centers as Lublin, Warsaw, Gdańsk, Poznań, Łódź, Silesia, Kraków and Toruń are presented, as well as the role of leaders who created theories and methodological studies on chromatography and related techniques. The role of sample preparation methods is emphasized based on the same or similar theoretical foundations. The directions of developing these techniques in Poland today and in the future are presented.