Simple and Comprehensive Two-Dimensional Reversed-Phase HPLC Using Monolithic Silica Columns

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• 作者：Nobuo Tanaka ; Hiroshi Kimura ; Daisuke Tokuda ; Ken Hosoya ; Tohru Ikegami ; Norio Ishizuka ; Hiroyoshi Minakuchi ; Kazuki Nakanishi ; Yukihiro Shintani ; Masahiro Furuno ; and Karin Cabrera
• 刊名：Analytical Chemistry
• 出版年：2004
• 出版时间：March 1, 2004
• 年：2004
• 卷：76
• 期：5
• 页码：1273 - 1281
• 全文大小：199K
• 年卷期：v.76,no.5(March 1, 2004)
• ISSN：1520-6882

文摘

Simple and comprehensive two-dimensional (2D)-HPLCwas studied in a reversed-phase mode using monolithicsilica columns for second-dimension (2nd-D) separation.Every fraction from the first column, 15 cm long (4.6-mm i.d.), packed with fluoroalkylsilyl-bonded (FR) silicaparticles, was subjected to the separation in the 2nd-Dusing one or two octadecylsilylated (C₁₈) monolithic silicacolumns (4.6-mm i.d., 3 cm). Monolithic silica columnsin the 2nd-D were eluted at a flow rate of up to 10 mL/min with separation time of 30 s that meets the fractionation every 15-30 s at the first dimension (1st-D)operated at a flow rate of 0.4-0.8 mL/min. Three caseswere studied. (1) In the simplest scheme of 2D-HPLC,effluent of the 1st-D was directly loaded into an injectorloop of 2nd-D HPLC for 28 s, and 2 s was allowed forinjection. (2) Two six-port valves each having a sampleloop were used to hold the effluent of the 1st-D alternatelyfor 30 s for one 2nd-D column to effect comprehensive2D-HPLC without the loss of 1st-D effluent. (3) Twomonolithic silica columns were used for 2nd-D by usinga switching valve and two sets of 2nd-D chromatographsseparating each fraction of the 1st-D effluent with the two2nd-D columns alternately. In this case, two columns ofthe same stationary phase (C₁₈) or different phases, C₁₈and (pentabromobenzyloxy)propylsilyl-bonded (PBB),could be employed at the 2nd-D, although the latterneeded two complementary runs. The systems producedpeak capacity of ~1000 in ~60 min in cases 1 and 2 andin ~30 min in case 3. The three stationary phases, FR,C₁₈, and PBB, showed widely different selectivity fromeach other, making 2D separations possible. The simpleand comprehensive 2D-HPLC utilizes the stability andhigh efficiency at high linear velocities of monolithic silicacolumns.