Università Campus-Biomedico, Centro Integrato di Ricerca, Roma, Italy.
Electrophoresis. 2012 Aug;33(16):2553-60. doi: 10.1002/elps.201200157.
Nano-LC and CEC were studied for the separation of cytochrome c tryptic digest. The peptides mixture was analyzed using either a nano-LC commercial or a laboratory assembled instrumentation coupled with an IT-ESI-MS by using a nanospray interface. CEC experiments were carried out with a CE apparatus coupled with the IT-ESI-MS through a liquid junction interface. Analytes were separated utilizing C18 silica based stationary phases, of different properties and origin, silica derivatized with cyano groups and C18 monolithic material. The last column, just because the chemical composition (absence of charged/chargeable groups) was tested only using nano-LC. Best results mainly related to the highest number of peptides separated and column equilibration time were obtained by nano-LC employing the C18 stationary phase (detection of 20 peptides, coverage of 88%). Similar results were achieved using both commercial and laboratory assembled instrumentation. The use of CEC revealed a higher separation efficiency and shorter analysis time. However, the number of separated peptides were lower than those observed in nano-LC. In CEC the use of capillaries packed with cyanosilica particles offered better results; however, less satisfactory than those observed in the miniaturized LC technique. Provided the use of the same stationary phase and taking into account the driving forces, the two techniques can be considered complementary, offering different information related to the retention times of the studied peptides.
纳米液相色谱和毛细管电泳（CE）用于细胞色素 c 酶切产物的分离。采用纳米喷雾接口，通过 IT-ESI-MS 联用，分别用商业纳米液相色谱和实验室组装的仪器对肽混合物进行分析。采用液体连接接口，通过 CE 仪器与 IT-ESI-MS 联用进行 CEC 实验。分析物采用不同性质和来源的 C18 硅胶固定相、氰基官能化硅胶和 C18 整体材料进行分离。最后一根柱子，仅仅因为化学成分（无带电/可带电基团），仅用纳米液相色谱进行了测试。最佳结果主要与分离的肽数量最多和柱平衡时间最短有关，纳米液相色谱采用 C18 固定相（检测到 20 个肽，覆盖率 88%）。使用商业和实验室组装的仪器均可获得类似的结果。使用 CEC 可提高分离效率和缩短分析时间。然而，分离的肽数量低于纳米液相色谱观察到的数量。在 CEC 中，使用填充氰基硅胶颗粒的毛细管可获得更好的结果；然而，不如在小型化液相色谱技术中观察到的结果令人满意。考虑到固定相的相同使用和驱动力，这两种技术可以被认为是互补的，提供了与研究肽的保留时间相关的不同信息。
