The invention relates to a content determination method for a spray for tendon and bone injury, in particular to effective ingredients paeoniflorin, icariin, ellagic acid, epimedin A, epimedin B and epimedin in the tendon and bone injury spray The content assay method of 7 kinds of compositions such as C, baohuoside I, comprises the preparation of need testing solution, the preparation of reference substance solution, HPLC method mensuration, adopts the content assay method provided by the invention, can measure multiple active ingredients simultaneously, The detection line is stable, the separation is good, the detection accuracy is high, and the stability is good, the content determination time is shortened, the detection efficiency is improved, and a relatively comprehensive quantitative detection method is provided for controlling the quality of the bone injury spray. The study on the stability of the tendons and bones injury spray provides a more comprehensive method for the quantitative study of the stability of the active ingredients.