Simple 2D‐HPLC using a monolithic silica column for peptide separation

Journal of Separation Science - Tập 27 Số 10-11 - Trang 897-904 - 2004
Hiroshi Kimura1, Tetsuya Tanigawa1, Hironobu Morisaka1, Tohru Ikegami1, Ken Hosoya1, N. Ishizuka2, Hiroyoshi Minakuchi2, Kazuki Nakanishi3, Mitsuyoshi Ueda4, Karin Cabrera5, Nobuo Tanaka
1Department of Polymer Science and Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan. Phone: +81 75 724 7809.
2Kyoto Monotech Co., 376-5-206 Tsukiyama-cho, Kuze, Minami-ku, Kyoto 601-8203, Japan
3Department of Material Chemistry, Graduate School of Engineering, Kyoto University, PRESTO, JST, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan
4Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan
5Merck, Darmstadt, D-64293, Germany

Tóm tắt

AbstractSeparation of peptides by fast and simple two‐dimensional (2D)‐HPLC was studied using a monolithic silica column as a second‐dimension (2nd‐D) column. Every fraction from the first column, 5 cm long (2.1 mm ID) packed with polymer‐based cation exchange beads, was subjected to separation in the 2nd‐D using an octadecylsilylated (C18) monolithic silica column (4.6 mm ID, 2.5 cm). A capillary‐type monolithic silica C18 column (0.1 mm ID, 10 cm) was also employed as a 2nd‐D column with split flow/injection. Effluent of the first dimension (1st‐D) was directly loaded into an injector loop of 2nd‐D HPLC. UV and MS detection were successfully carried out at high linear velocity of mobile phase at 2nd‐D using flow splitting for the 4.6 mm ID 2nd‐D column, or with direct connection of the capillary column to the MS interface. Two‐minute fractionation in the 1st‐D, 118‐second loading, and 2‐second injection by the 2nd‐D injector, allowed one minute for gradient separation in the 2nd‐D, resulting in a maximum peak capacity of about 700 within 40 min. The use of a capillary column in the 2nd‐D led to less solvent consumption and better MS detectability compared to a larger‐sized column. This kind of fast and simple 2D‐HPLC utilizing monolithic silica columns will be useful for the separation of complex mixtures in a short time.

Từ khóa


Tài liệu tham khảo

L.Mondello A.C.Lewis K.D.Bartle Multidimensional Chromatography John Wiley & Sons Ltd. Chichester England 2002.

10.1002/(SICI)1521-4168(20000301)23:3<167::AID-JHRC167>3.0.CO;2-2

10.1021/ac000578r

10.1021/ac00201a015

10.1021/ac00114a026

10.1021/ac961155l

10.1021/ac961156d

Wehr T., 2002, LC‐GC, 20, 954

10.1007/BF02467508

10.1016/S0021-9673(99)00206-X

10.1016/S0021-9673(00)00736-6

10.1002/(SICI)1521-4168(20000301)23:3<259::AID-JHRC259>3.0.CO;2-V

10.1021/ac010627f

10.1021/ac020534d

10.1016/S0378-4347(00)00547-8

10.1021/ac60252a025

10.1021/ac60293a001

10.1021/ac9807013

10.1021/ac980702v

10.1021/ac010617e

10.1021/ac034925j

10.1021/ac960281m

10.1016/S0021-9673(97)00947-3

10.1021/ac012495w

10.1016/S0021-9673(01)01582-5

10.1016/S0021-9673(02)00133-4

10.1016/S0021-9673(01)81640-X

Thông tin
Thông tin xuất bản

Journal of Separation Science

Tập 27 Số 10-11

897-904

Thông tin tác giả