Vydac 214TP C4 Reversed-Phase Columns for …

1 DescriptionVydac C4 ( 214TP ) Reversed-Phase material consistsof butyl aliphatic groups bonded to the surface of300 pore diameter silica. The large pores of the300 TP silica allow polypeptide molecules completeaccess to the interior of the silica pores. The uniqueprocess by which we manufacture Vydac TP silicaresults in a very high purity synthetic silica withcarefully controlled characteristics. Vydac TP silica isthe standard that has defined large pore HPLC silicafor polypeptide separations for nearly two decades(see Ref. 1 on page 11).The hydrophobic (reversed) phase is attached to thesilica using polyfunctional chlorobutylsilanes, resultingin cross-linking, or polymerization, of the hydrophobicphase.

2 The practical benefits of the polymeric phaseare:n very long column lifetimen no measurable phase leachingCarbon loading and end-capping are closely monitoredby applications-focused quality control tests of theseparation of a selected set of 214 ATP54 is a special column for which theadsorbent chemistry is optimized for use in stabilityanalysis of human growth hormone. (See chromatogramand Ref. 4 on page 23.) Polymeric bonding results in excep-tionally long column lifetime andnegligible phase leaching. 300 silica allows full access to thereversed-phase surface by evenlarge polypeptides. Applications-focused quality controlassures highest level of performanceand reproducibility.

3 Now available in Microbore, Narrowbore, and 214TP C4 Reversed-Phase Columnsfor PolypeptidesApplicationsVydac C4 Reversed-Phase materials are typicallyused and recommended for the separation of: Glycoproteins (Ref. 1) Hemoglobin variants (Ref. 2) Histones (Ref. 3) Human Growth hormone (Ref. 4) Insulin variants (Ref. 5) Membrane proteins (Ref. 6)(All references above refer to listing on page 23.)Cat. Analytical Sizes214 ATP54 Column, C4, 300 , 5 m, mm x 250 mm214TP54 Column, C4, 300 , 5 m, mm x 250 mm214TP5415 Column, C4, 300 , 5 m, mm x 150 mm214TP5405 Column, C4, 300 , 5 m, mm x 50 mmSolvent-Saver Analytical214TP53 Column, C4, 300 , 5 m, mm x 250 mm214TP5315 Column, C4, 300 , 5 m, mm x 150 mmSemipreparative214TP510 Column, C4, 300 , 5 m, 10 mm x 250 mmOther column sizes and particle diameters are available forboth analytical and preparative applications.

4 Please consultthe selection guide on page 32 for additional listings. 214TP is a trademark of The Separations Group, InformationReferences1. Microscale Structure Analysis of a high Molecular WeightHydrophobic Membrane Glycoprotein Fraction with Platelet-Derived Growth Factor-Dependent Kinase Activity, P. Tempst,D. Woo, D. Teplow, R. Aebersold, L. Hood, and S. Kent, J. , 403-412 (1986).2. high Performance Liquid Chromatographic Separation of GlobinChains on a Large-Pore C4 Column, Shelton, Shelton,and Schroeder, J. Liq. Chrom. 7(10), 1969-1977 (1984).3. Separation and Purification of S49 Mouse Lymphoma Histonesby Reversed-Phase high -Performance Liquid Chromatography,M.

5 C. McCroskey, Groppi, and Pearson, Anal. , 427-432 (1987).4. A Reversed-Phase high Performance Liquid ChromatographicMethod for Characterization of Biosynthetic Human GrowthHormone, Riggin, Dorulle, and Miner, Anal. Biochem. 167, 199-209 (1987).5. Reversed-Phase high -Performance Liquid Chromatography ofInsulins from Different Species, J. Rivier and R. McClintock,J. Chrom. 268, 112-119 (1983).6. Purification of Integral Plasma Membrane Proteins by Reverse-Phase high Performance Liquid Chromatography, Sussman,Anal. Biochem. 169, 395-399 (1988). Vydac C4 Reversed-Phase Columns for PolypeptidesRibosomal proteinsLarge, hydrophobic proteins are best separated on C4reversed-phase Columns .

6 While suitable for peptides as smallas 10-20 residues, the short, mildly hydrophobic C4 phasegives especially good recovery and resolution for large orhydrophobic : Vydac 214TP54 (C4, 5 m, mm x 250 mm).Eluent: 10-38% isopropanol with TFA over 355 Kamp, A Rossenthoff, D. Kamp, and B. Wittman-Liebold,J. Chrom 317, 181-192 (1984).360 min0mono-desamidoHGHH uman Growth HormoneHuman growth hormone is routinely assayed for stability by measuringdegradation products. Both deamidation and oxidation products areresolved by isocratic Reversed-Phase : Vydac 214 ATP54 (C4, 5 m, mm x 250 mm L).Eluent: 29% isopropanol/71% M Tris buffer, pH from Riggin, Dorulla, and Miner, Anal.

7 Biochem. 167,199-209 (1987).(Note: The use of an eluent at pH will decrease the column lifetime and is notgenerally recommended.)human growth hormone(HGH)42 min0di-desamidoHGH214 ATP54