Elution profile of di-peptides on a sulfonated ethylstyrene-divinylbenzene copolymer resin column by high-performance liquid chromatography

Jian Guo, Tomomi Saiki, Kumrungsee Thanutchaporn, Wanying Liu, Akihiro Shimura, Toshiro Matsui

Research output: Contribution to journalArticle

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Abstract

This study investigates the characteristics of a partially sulfonated ethylstyrene-divinylbenzene copolymer for the separation of di-peptides by high-performance liquid chromatography. Di-peptides (VE, VA, VH, VK, and VR) with different isoelectric points (pI, 4.0 to 9.7) and log P values (-1.63 to -0.72) were used to optimize the separation conditions of the columns packed with sulfonated copolymer resin. The retention factor (k) of the di-peptides on the column with a 0.81 wt% sulfo group content decreased with increasing concentrations of phosphate salts (2.5-20 mmol L -1 ) in the mobile phase. The complete separation of the five di-peptides was obtained with a gradient of 10% methanol containing 5 mmol L -1 NaH 2 PO 4 (pH 4.8) to 50% methanol containing 5 mmol L -1 NaH 2 PO 4 (pH 8.9) for 60 min at 0.5 mL min -1 at 50°C. Under the optimal conditions, a good relationship between the k and pI values of the di-peptides, with the exception of VE (pI 4.0), was observed, suggesting that the retention of the di-peptides on the column packed with sulfonated copolymer resin was dependent on the pI value, when it was greater than 5. The log P value also influenced the separation characteristics of the column; peptides possessing the same pI value (6.4 for GH, VH, IH, and FH) showed a higher retention on the column with increasing log P values. In conclusion, the prototype sulfonated ethylstyrene-divinylbenzene copolymer column was applicable for the separation of basic di-peptides, and the separation depended on the pI and hydrophobicity of the di-peptides. 2014.

Original languageEnglish
Pages (from-to)45-50
Number of pages6
Journalanalytical sciences
Volume31
Issue number1
DOIs
Publication statusPublished - Jan 1 2015

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divinyl benzene
High performance liquid chromatography
Copolymers
Resins
Peptides
Methanol

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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Elution profile of di-peptides on a sulfonated ethylstyrene-divinylbenzene copolymer resin column by high-performance liquid chromatography. / Guo, Jian; Saiki, Tomomi; Thanutchaporn, Kumrungsee; Liu, Wanying; Shimura, Akihiro; Matsui, Toshiro.

In: analytical sciences, Vol. 31, No. 1, 01.01.2015, p. 45-50.

Research output: Contribution to journalArticle

Guo, Jian ; Saiki, Tomomi ; Thanutchaporn, Kumrungsee ; Liu, Wanying ; Shimura, Akihiro ; Matsui, Toshiro. / Elution profile of di-peptides on a sulfonated ethylstyrene-divinylbenzene copolymer resin column by high-performance liquid chromatography. In: analytical sciences. 2015 ; Vol. 31, No. 1. pp. 45-50.
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abstract = "This study investigates the characteristics of a partially sulfonated ethylstyrene-divinylbenzene copolymer for the separation of di-peptides by high-performance liquid chromatography. Di-peptides (VE, VA, VH, VK, and VR) with different isoelectric points (pI, 4.0 to 9.7) and log P values (-1.63 to -0.72) were used to optimize the separation conditions of the columns packed with sulfonated copolymer resin. The retention factor (k) of the di-peptides on the column with a 0.81 wt{\%} sulfo group content decreased with increasing concentrations of phosphate salts (2.5-20 mmol L -1 ) in the mobile phase. The complete separation of the five di-peptides was obtained with a gradient of 10{\%} methanol containing 5 mmol L -1 NaH 2 PO 4 (pH 4.8) to 50{\%} methanol containing 5 mmol L -1 NaH 2 PO 4 (pH 8.9) for 60 min at 0.5 mL min -1 at 50°C. Under the optimal conditions, a good relationship between the k and pI values of the di-peptides, with the exception of VE (pI 4.0), was observed, suggesting that the retention of the di-peptides on the column packed with sulfonated copolymer resin was dependent on the pI value, when it was greater than 5. The log P value also influenced the separation characteristics of the column; peptides possessing the same pI value (6.4 for GH, VH, IH, and FH) showed a higher retention on the column with increasing log P values. In conclusion, the prototype sulfonated ethylstyrene-divinylbenzene copolymer column was applicable for the separation of basic di-peptides, and the separation depended on the pI and hydrophobicity of the di-peptides. 2014.",
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