Gabriel T F, Jimenez M H, Felix A M, Michalewsky J, Meienhoffer J
Int J Pept Protein Res. 1977;9(2):129-36. doi: 10.1111/j.1399-3011.1977.tb03472.x.
A simple preparative system is described for rapid and efficient purification of protected synthetic peptides on a gram scale by high performance liquid chromatography on prepacked silica gel 60 columns. A variety of protected peptides up to tetradecapeptides have been chromatographed at pressures of 50 to 150 psi and obtained in analytically pure from within 2 to 4 h. With such commonly used protecting groups as N-benzyloxycarbonyl (Z), N-2-(p-biphenylyl)-2-propyloxycarbonyl (Bpoc), N-t-butyloxycarbonyl (Boc), O- and S-t-butyl (But), and S-acetamidomethyl (Acm), compounds were sufficiently soluble in chloroform, alcohols, acetic acid, or mixtures of these solvents for column loading. Dimethylformamide was also used as a solvent for loading. Solvent systems for column elution in isocratic, stepwise, or gradient modes were composed of chloroform, isopropanol, ethanol, or methanol and acetic acid in ratios that differed for each protected peptide depending on Rf values on t.l.c. plates. A simple chromatography is described which was self-assembled using standard instruments commonly in use in most laboratories. A shut-off valve was designed to prevent loss of material between fractions.
本文描述了一种简单的制备系统，用于在预装硅胶60柱上通过高效液相色谱法快速高效地纯化克级规模的受保护合成肽。多种高达十四肽的受保护肽已在50至150 psi的压力下进行色谱分离，并在2至4小时内获得分析纯品。使用诸如N-苄氧羰基（Z）、N-2-（对-联苯基）-2-丙氧羰基（Bpoc）、N-叔丁氧羰基（Boc）、O-和S-叔丁基（But）以及S-乙酰氨基甲基（Acm）等常用保护基团时，化合物在氯仿、醇类、乙酸或这些溶剂的混合物中具有足够的溶解性以用于柱上负载。二甲基甲酰胺也用作负载溶剂。用于等度、逐步或梯度模式柱洗脱的溶剂系统由氯仿、异丙醇、乙醇或甲醇与乙酸组成，其比例因每种受保护肽而异，具体取决于薄层层析板上的比移值。本文描述了一种使用大多数实验室常用的标准仪器自行组装的简单色谱法。设计了一个截止阀以防止各馏分之间的物料损失。
