Dipartimento Farmaco-chimico, Facoltà di Farmacia, Università di Messina, Messina, Italy.
J Sep Sci. 2012 Feb;35(4):530-3. doi: 10.1002/jssc.201100877.
In the present contribution, a fully automated capillary comprehensive two-dimensional LC (LC×LC) method, for proteomic analysis, was developed for the first time. The investigated platform was characterized by the coupling of high-pH RP with low-pH RP separations thus ensuring the generation of high peak capacity despite the employment of identical stationary phases. The use of capillary columns in both dimensions allowed to reduce mobile-phase consumption and enhance sensitivity. Fraction transfer from the first to the second dimension was performed by means of two 2-position 6-port nano-switching valves, under stop-flow conditions. Values as high as 1208 and 955 were obtained for the theoretical and practical peak capacity, respectively. The investigated LC×LC system showed good retention time repeatibility with RSD values ranging from 0.8 to 6.0% for the first dimension and from 1.0 to 3.0% for the second dimension, respectively. RSD peak area values below 9.5% were also attained, thus demonstrating the precision of the LC×LC method employed.
在本研究中，首次开发了一种完全自动化的毛细管二维 LC（LC×LC）方法，用于蛋白质组学分析。该研究平台的特点是采用高 pH 值反相和低 pH 值反相分离相结合，从而确保在使用相同固定相的情况下产生高的峰容量。在两个维度上均使用毛细管柱，可减少流动相消耗并提高灵敏度。通过两个 2 位置 6 端口纳米切换阀，在停流条件下实现第一维到第二维的馏分转移。理论和实际的峰容量分别高达 1208 和 955。所研究的 LC×LC 系统显示出良好的保留时间重现性，第一维的 RSD 值范围为 0.8%至 6.0%，第二维的 RSD 值范围为 1.0%至 3.0%。还获得了低于 9.5%的 RSD 峰面积值，从而证明了所采用的 LC×LC 方法的精度。
