RP-HPLC determination of recombinant Escherichia coli fermentation broth of the content of antihypertensive peptides.doc

 PAGE \* MERGEFORMAT 10 RP-HPLC determination of recombinant Escherichia coli fermentation broth of the content of antihypertensive peptides Author: Liao Xiaohui Sun Haiyan Liu Peng Guanghua Zhang Shenghua 【Abstract】 Objective To establish a reversed-phase high performance liquid chromatography (RP  HPLC) determination of recombinant Escherichia coli fermentation broth method of antihypertensive peptides, their in-depth study provides methodological basis. Methods Aydac C18 reversed-phase column (4.6 mm × 250 mm, 5 μm), Aydac guard column, column temperature 30 ℃, with acetonitrile: water (22:78), 0.05% TFA as mobile phase, flow rate 1.0 ml / min The detection wavelength was 215 nm, reference wavelength 350 nm, injection volume 20 μl. The results of the method makes antihypertensive peptides and other similar products are well separated, at 5 ~ 250 μg / ml a good linear relationship, r = 0.999 8, the minimum detection limit of 8 ng, low, medium and high concentration increases in sample recovery rate, days, day precision requirements are in line with the methodology. Conclusion The method is stable, high sensitivity, easy operation, suitable for re-determination of antihypertensive peptides. Keywords: recombinant Escherichia coli; antihypertensive peptides; RP-HPLC Abstract: ObjectiveA simple and rapid reversed-phase high performance liquid chromatographic method was developed for the AHP in Recombinant E.coli fermentation broth.MethodsThe separation conditions were: Aydac C18 reversed phase column, Aydac guard column. Mobile phase was acetonitrile: water (22 : 78), 0.05% TFA, flow rate 1ml/min.The detection wavelength was 215nm, reference wavelength was 350 nm.Column temperature was 30 ℃. Injection volume: 20 μl. ResultsAHP has good separation from other components in fermentation broth. And there was perfect linear correlation between 5 ~ 250 μg / ml, r = 0.999 8. The limit of detection was 8ng. The intra-and inter-assay precisio