The invention relates to an isopentene flavonol glycoside derivative with a structure shown as a general formula (I) and a preparation method and application thereof. In the isopentene flavonol glycoside derivative, R1 refers to rhamnosyl, R2, R3, R4 and R7 refer to H, R5 refers to methoxyl, and R6 refers to hydroxyl; or R1, R4, R6 and R7 refer to H, R2 refers to xylosyl, R3 refers to acetyl, and R5 refers to hydroxyl; or R1, R3, R6 and R7 refer to H, R2 refers to xylosyl, R4 refers to glucosyl, and R5 refers to hydroxyl; or R1 refers to rhamnosyl, R2, R3 and R6 refer to H, R4 refers to glucosyl, R5 refers to methoxyl, and R7 refers to hydroxyl. The invention further relates to application of the isopentene flavonol glycoside derivative to the prevention and treatment of osteoporosis.