Superiority of a new gradient system utilizing a critical threshold for the adsorption capacity of polypeptides to column packing when compared with a standard gradient system for the simultaneous and quantitative analysis of polypeptides.
Compared with a standard gradient system, the new gradient system which we developed has a major advantage because it permits a wide range of acetonitrile content, e.g. more than the critical threshold, in the polypeptide solution and allows the quantitative analysis of the polypeptide with satisfactory analytical precision. Additionally, this new gradient system allows the enhancement of the sensitivity of the polypeptide analysis proportionate to the increased volume of solution loaded with the same levels of precision. In contrast, when using a standard gradient system it is difficult to analyze a polypeptide quantitatively with good precision due to either adsorption to various materials or to irregular change in the ratio between a retained and a passed peak of the polypeptide. Additionally, the appearance of a passed peak results in a loss in the sensitivity of the polypeptide analysis, although no adsorption of a polypeptide to various materials occurs in a solution with acetonitrile content more than the critical threshold. Consequently, the new gradient system is effective for the simultaneous and quantitative analysis of different polypeptides with good precision and without any loss of sensitivity due to either adsorption to various materials or the appearance of a passed peak.
Goda
Ryoya
R
Drug Metabolism and Pharmacokinetics Research Laboratories, R&D Division, Daiichi Sankyo Co. Ltd, 1-2-58, Hiromachi, Shinagawa-ku, Tokyo, Japan. goda.ryoya.u4@daiichisankyo.co.jp
Masumoto
Hiroshi
H
Okazaki
Osamu
O
Sudo
Kenichi
K
D000327:Adsorption; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods; D010455:Peptides / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D054832:Urocortins / Q000032:analysis
D010455:Peptides; D054832:Urocortins