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Analysis and Preparation of Linaclotide by High Performance Liquid Chromatography
Yi-Shen Zhu, Tao Feng, Yong-Qin Diao and Chun-Yan Tu
Linaclotide, a synthetic peptide, is a guanylate cyclase-C (GC-C) agonist for treating irritable bowel syndrome with constipation and chronic idiopathic constipation. A simple, precise and selective RP-HPLC method for linaclotide analysis was developed and validated. Analysis was achieved with a SinoChrom ODS-BP column, mobile phase A and B comprised 30 mol·L-1 phosphate (pH 2.8) solution and acetonitrile respectively, gradient elution B% set to 7%?25% from 0 to 32 min and to 25% from 32 to 50 min, flow rate of 1.0 mL·min-1, oven temperature at 40?, injection volume of 20 µL, and UV detection wavelength at 214 nm. Method validation was carried out in linearity, LOD, LOQ, precision and stability. Modified method for linaclotide preparation on a preparative liquid chromatography was developed from the analytical method and applied successfully.