The present invention provides an alkynyl multi-arm polyethylene glycol derivative having a structure of a general formula I or general formula X VIII. In the derivative, X1, X2, X3 and X4 are linking groups, F1, F2, F3 and F4 are end groups, the end groups may be the same or may also be different, and are selected from: hydroxy, carboxyl, ester group, amino, alkynyl or the like, at least one of the end groups is alkynyl, and PEG is the same or different —(CH2CH2O)m—, wherein m is an integer ranging from 3 to 250, and l is an integer greater than or equal to 1. The multi-arm polyethylene glycol derivatives have stronger application flexibility, and have good application prospect in aspects such as organic synthesis, medicine synthesis and medical instruments.