The present invention relates to the field of surfactants, particularly to a sorbitan fatty acid ester glycoside preparation method, which comprises: pouring a span-based compound and glucose into the same reactor, carrying out a dehydrating condensation reaction in the presence of an acidic catalyst to form a glycosidic bond, adding an alkali after completing the reaction, neutralizing to obtain a span polyglycoside crude product, and separating to obtain the glycoside represented by the following formula. According to the present invention, the preparation method has advantages of simple process, green and environmentally friendly raw materials, and low cost; the water solubility problem of the span series products can be effectively improved, the application range of the span series products can be expanded, and the sorbitan fatty acid ester glycoside is the green surfactant, and most likely becomes the substitute for the tween series products. The formula is defined in the specification.