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a) dissolving crude Semaglutide backbone in a buffer solution, b) subjecting the solution to RP-HPLC purification using buffer solution as mobile phase A and acetonitrile optionally in mixture with alcohol solvent as mobile phase B, c) subjecting the solution to RP-HPLC purification by using buffer solution as mobile phase A and acetonitrile as mobile phase B, d) subjecting the solution to RP-HPLC by using water as mobile phase A and acetonitrile as mobile phase B