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. Author manuscript; available in PMC: 2009 Nov 6.
Published in final edited form as: J Sep Sci. 2008 Aug;31(15):2754–2773. doi: 10.1002/jssc.200800243

Figure 14.

Figure 14

Analysis and purification of synthetic peptides by RPC and HILIC/CEX. (A) 35-residue Cys-containing amphipathic α-helical peptide; (B) 17-residue intrachain disulphide-bridged peptide. Columns: RPC and HILIC/CEX (denoted HILIC/CEC in Fig. 14), same as Fig. 3. Conditions: RPC, linear AB gradient (1% B/min, equivalent to 1% ACN/min) at a flow-rate of 1 mL/min, where eluent A is 0.05% aq. TFA and eluent B is 0.05% TFA in ACN; HILIC/CEX, linear AB gradient (2.5 mM NaClO4/min from 30 mM NaClO4, following 10-min isocratic elution with 30 mM NaClO4) at a flow-rate of 1 mL/min, where eluent A is 5 mM aq. TEAP, pH 7, and eluent B is 5 mM aq. TEAP containing 400 mM NaClO4, pH 7, both eluents also containing 65% v/v ACN; runs carried out at 26°C and peaks detected by absorbance at 210 nm. Peptide sequences: (A) Ac-Gln-Cys-Gly-Ala-Leu-Gln-Lys-Gln-Val-Gly-Ala-Leu-Glu-Lys-Glu-Glu-Gly-Ala-Leu-Glu-Lys-Gln-Val-Gly-Ala-Leu-Gln-Lys-Gln-Val-Gly-Ala-Leu-Gln-Lys-amide; (B) Ac-Lys-Cys-Lys-Ser-Thr-Gln-Asp-Glu-Gln-Phe-Ile-Pro-Lys-Gly-Cys-Ser-Lys (intrachain disulphide bridge between two Cys residues; Ac denotes Nα-acetyl and amide denotes Cα-amide). Reprinted from ref. [6] with permission of Academic Press.