The invention provides a preparation method for an alkynyl-containing steroid nucleus derivative. The method comprises the following three steps of first replacement reaction, second replacement reaction and extraction. The alkynyl-containing steroid nucleus derivative with high purity can be prepared by using the method. In the method, an alkali compound is used for preparing alkyne salt in an aprotic polar solvent, the coordination of the aprotic polar solvent and the alkali compound are used for increasing the alkalinity of the alkali compound exponentially and effectively increasing the solubility of the ethyne, and the activity of the generated alkyne salt is strengthened because of the adding of the aprotic polar solvent. In addition, the method has the characteristics of high reaction efficiency, moderate reaction condition, good selectivity, high yield, and convenient operation. Compared with the prior art, the preparation method has the obvious economic benefit.