High-performance liquid peak capacity

Gilar, M., Daly, A.E., Kele, M., Neue, U.D., Gebler, J.C. (2004). Implications of column peak capacity on the separation of complex peptide mixtures in single- and two-dimensional high-performance liquid chromatography. J. Chromatogr. A 1061, 183-192. 

Fig. 3 Chromatograms obtained by pH zone-refining CCC. (A) Separation of CBZ(Z) dipeptides. Experimental conditions are as follows apparatus type-J multilayer CPC (PC Inc., Potomac, MD, USA) with a 10-cm revolution radius column multilayer coil, 1.6-mm ID, 325 mL capacity solvent system methyl tert-h xiy ether/acetonitrile/water (2 2 3), 16 mM TEA in organic phase (pH 1.83), and 5.5 mM NH3 in aqueous phase (pH 10.62) sample eight CBZ(carbobenzyloxy) dipeptides as indicated in the chromatogram, each 100 mg in 50 mL of solvent (25 mL in each phase) flow rate 3.3 mL/hr in head-to-tail elution mode detection 206 nm revolution 800 rpm retention of stationary phase 65.1%. (B) Separation of bacitracin complex. High-performance liquid chromatography (HPLC) analysis indicated that two major components, bacitracins A and B, were isolated in peaks 3 and 5, respectively. Experimental conditions are as follows apparatus and column same as above solvent system methyl r -butyl ether/ acetonitrile/water (4 1 5), 40 mM triethylamine, 10% DEHPA in the organic stationary phase, and 20 mM HCl in aqueous mobile phase flow rate 3 mL/min sample 5 g of bacitracin dissolved in 40 mL of solvent (20 mL in each phase) revolution 800 rpm detection 280 nm.

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