The invention relates to a preparation method of a chitosan hydroxybutyl derivative. Chitosan is used as a raw material; 1,2-epoxybutane is used as an etherifying agent; the hydroxybutyl modification of the chitosan is performed in an isopropanol aqueous solution dispersion system under microwave action with controlled reaction temperature and time; a chitosan derivative, hydroxybutyl chitosan, is successfully prepared; the preparation period is greatly shortened, and the preparation efficiency is increased. During the reaction, the hydroxybutyl modification of the chitosan is performed orderly under the microwave action and with controlled reaction temperature and time, which shortens the preparation period. The product has a wider pH dissolution range, and has a soluble property in weakacid, neutral, and weak alkaline aqueous solutions, that is to say, the limitation that chitosan is only soluble in acids or acid solutions is improved; the aqueous solution of the product has temperature sensitivity, can be transformed between an aqueous solution and hydrogel by adjusting the temperature; and the product is widely applicable to fields of biomedicine, tissue engineering, pharmaceutical carriers, chemical engineering, food, cosmetics, etc.