Wilson K J, Honegger A, Hughes G J
Biochem J. 1981 Oct 1;199(1):43-51. doi: 10.1042/bj1990043.
The chromatographic properties of peptides ranging in length from 2 to 65 residues have been compared on reverse-phase packings in three buffer systems at low pH. Of the buffers examined, two are widely used in connection with u.v. detection [(i) phosphate/acetonitrile or (ii) phosphate/propan-2-ol] and the third for fluorescence detection [(iii) pyridine/formate-pyridine/acetate/propan-1-ol]. The addition of a chaotropic salt, NaClO4, to the phosphate buffers, as first described by Meek (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 1632-1636, is shown to significantly improve the chromatographic behaviour of more hydrophobic peptides. The two most commonly used detection systems, u.v. and fluorescence, are compared in terms of ease of use and sensitivity.
在低pH值下,已在三种缓冲系统中,于反相填料上比较了长度范围为2至65个残基的肽的色谱特性。在所研究的缓冲液中,两种广泛用于紫外检测[(i)磷酸盐/乙腈或(ii)磷酸盐/丙-2-醇],第三种用于荧光检测[(iii)吡啶/甲酸盐-吡啶/乙酸盐/丙-1-醇]。如Meek(1980年,《美国国家科学院院刊》77, 1632 - 1636)首次所述,向磷酸盐缓冲液中添加离液盐高氯酸钠,可显著改善疏水性更强的肽的色谱行为。就易用性和灵敏度而言,对两种最常用的检测系统,即紫外检测和荧光检测进行了比较。