Newly Devised Peptide Chiral Stationary Phases Evaluated for Enantioselectivity in HPLC Applications

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Researchers from Hebei University in China have developed and characterized three novel peptide chiral stationary phases (CSPs) for enantiomeric separation, demonstrating their potential in high performance liquid chromatography (HPLC).

In a recent study published in Chromatographia, a team of researchers from the Key Laboratory of Pharmaceutical Quality Control of Hebei Province in the College of Pharmaceutical Sciences at Hebei University in Baoding, China introduced three new peptide chiral stationary phases (CSPs) with promising enantioselective capabilities (1). These phases, designed with a main chain comprised of L-proline (L-Pro), L-phenylalanine (L-Phe), L-valine (L-Val), and L-leucine (L-Leu), offer new opportunities in the field of high performance liquid chromatography (HPLC) for resolving chiral compounds. This development addresses a need in both the pharmaceutical and analytical chemistry industries.

The researchers crafted these stationary phases by immobilizing L-citrulline, L-lysine, and L-tryptophan on 3-aminopropyltrimethoxysilane-modified silica gel (1). To ensure their quality and reliability, the newly developed phases underwent comprehensive analyses and characterizations, including Fourier transform infrared (FT-IR) spectra, elemental analysis, and thermogravimetric analysis.

One of the primary objectives of this study was to evaluate the enantioselective performance of the three peptide stationary phases. This evaluation involved nine racemic compounds and was conducted under normal-phase HPLC conditions (1). The researchers optimized the enantiomeric separation conditions and achieved what they described as impressive results. For instance, on the CSP-1 column, the resolutions of compounds such as flurbiprofen, naproxen, benzoin, 1,1ʹ-bi-2-naphthol, and ketoprofen were measured at 1.61, 2.0, 0.62, 0.52, and 1.20, respectively. These results demonstrated the potential of these novel stationary phases for separating a wide range of chiral compounds effectively.

Furthermore, the study investigated the reproducibility of the CSP-1 column, a crucial factor in chromatography. The findings indicated that these peptide stationary phases exhibit reproducibility with a relative standard deviation (RSD) of just 0.12% (n=5) (1).


The significance of this research lies in its contribution to the field of chromatography, particularly in the realm of chiral separations. Chiral compounds, which exist as mirror-image isomers, often possess distinct biological activities, making their separation a fundamental requirement in pharmaceutical development and chemical analysis. The introduction of these novel peptide chiral stationary phases represents a promising advancement in this domain.

The development and characterization of the L-Pro-L-Phe-L-Val-L-Leu peptide stationary phases exemplifies continuous efforts to advance analytical techniques and contribute to the broader understanding of complex chemical systems.


(1) Guo, X.; Shang, P.; Wei, B.; et al. Preparation and Enantiomeric Separation of L-Pro-L-Phe-L-Val-L-Leu Peptide Stationary Phases. Chromatographia 2023, 86, 541–551. DOI: 10.1007/s10337-023-04267-y