Zheng Zhaobin, Ying Wantao, Cai Yun, Tian Zhongmin, Qian Xiaohong
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation, Beijing 102206, China.
Se Pu. 2007 Nov;25(6):804-8.
On the basis of theoretical pH calculation for buffer systems, an off-line linear pH gradient-strong cation exchange chromatographic method was developed for peptides separation. For the acetate buffer system, the peptides from tryptic digest of BSA were eluted by the linear pH gradient (pH 3.7-6.0) with a low concentration of ammonium acetate salt gradient, whose salt is volatile and can be easily removed by lyophilization. As for the citrate buffer system, the peptides were eluted by a wider range of linear pH gradient (pH 3.0-8.5) with a even lower concentration of ammonium citrate salt gradient. Both methods are effective for the peptides separation and before mass spectrometric detection can simplify the desalting step, which is labor-intensive and may incur significant sample loss for the routine strong cation exchange chromatography.
基于缓冲体系的理论pH计算，开发了一种离线线性pH梯度-强阳离子交换色谱法用于肽段分离。对于醋酸盐缓冲体系，牛血清白蛋白胰蛋白酶消化产生的肽段通过线性pH梯度（pH 3.7 - 6.0）和低浓度醋酸铵盐梯度进行洗脱，该盐具有挥发性，可通过冻干轻松去除。至于柠檬酸盐缓冲体系，肽段通过更宽范围的线性pH梯度（pH 3.0 - 8.5）和更低浓度的柠檬酸铵盐梯度进行洗脱。这两种方法对肽段分离均有效，且在质谱检测前可简化脱盐步骤，而常规强阳离子交换色谱的脱盐步骤 labor-intensive 且可能导致大量样品损失。 （注：原文中“labor-intensive”未翻译，可能是你提供的原文有误，该词意为“劳动密集型的” ）
