First, each sample (30.00 mg) was weighed and transferred to a 2 mL centrifuge tube, followed by adding two 5 mm magnetic beads, an appropriate amount of SDS-containing protein lysis buffer, and 1× Cocktail (protease inhibitor) with a final concentration of EDTA. The samples were disrupted and lysed using an automatic grinder, then centrifuged at 25,000g at 4℃ for 15 minutes to collect the supernatant. Next, DTT was added to the supernatant to a final concentration of 10 mM, and the mixture was incubated in a 37℃ water bath for 30 minutes. Subsequently, IAM was added to a final concentration of 55 mM, and the samples were placed in a dark room for 45 minutes. Five volumes of pre-cooled acetone were then added, and the mixture was stored in a -20℃ refrigerator for 2 hours, followed by centrifugation at 25,000g at 4℃ for 15 minutes to discard the supernatant. The precipitate was air-dried, and an appropriate amount of SDS-free protein lysis buffer was added. An automatic grinder was used to promote protein dissolution, and the mixture was centrifuged again at 25,000g at 4℃ for 15 minutes. The resulting supernatant was the final protein solution, which was used for subsequent quantification and analysis.