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A process for the purification of Semaglutide backbone (H-His-Aib-Glu-Gly-Thr-Phe- Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-IIe-Ala-Trp-Leu- Val-Arg-Gly-Arg-Gly-OH) comprising, subjecting a solution of Semaglutide backbone to RP-HPLC purification by using buffer solution as mobile phase, wherein the buffer solution is selected from aqueous ammonia, aqueous ammonium chloride, aqueous ammonium acetate, aqueous ammonium bicarbonate, aqueous sodium acetate, aqueous sodium chloride, aqueous sodium carbonate, aqueous sodium bicarbonate, aqueous monosodium phosphate, aqueous disodium phosphate, aqueous KH 2 PO4, aqueous tris(hydroxymethyl)aminomethane (Tris buffer), aqueous formic acid, aqueous acetic acid, citric acid in water, aqueous boric acid, aqueous sulfuric acid, isopropyl methylphosphonic acid in water, O -phthalaldehyde (OPA) in water or mixtures thereof

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