The invention discloses a semi-aqueous cleaning composition and a preparation method and application thereof. The composition comprises 5-20% of a permeation solvent, 1-5% of surface active peptide, 2-10% of sophorolipid, 1-5% of NaOH, 1-5% of sodium carbonate, 3-8% of sodium metasilicate, 2-15% of N, N-diethylhydroxylamine and water. The osmotic solvent is one or two of aldehyde condensation 2, 3-butanediol and ketone condensation 2, 3-butanediol, and the osmotic solvent is one or two of aldehyde condensation 2, 3-butanediol and ketone condensation 2, 3-butanediol; the mold cleaning agent for resin lens production, provided by the invention, has small damage to a mold, has a good removal effect on residual adhesives, resin monomers and resin solids at the edge of the mold, and effectively improves the qualified rate of resin lens production. Meanwhile, the cleaning agent provided by the invention has the characteristics of thorough oil removal, dust removal, surface residual fiber removal and the like, and the qualified rate of resin lens coating is effectively improved.