Reversed-phase fused-core HPLC modeling of peptides

D׳Hondt, Matthias; Gevaert, Bert; Stalmans, Sofie; Dorpe, Sylvia Van; Wynendaele, Evelien; Peremans, Kathelijne; Burvenich, Christian; Spiegeleer, Bart De

doi:10.1016/j.jpha.2012.11.002

Cited by 14 publications

(6 citation statements)

References 62 publications

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“…Besides the commonly used C18 fused-core columns, where separation efficiency under different conditions was investigated for a wide variety of chemical components [11] , [15] , [16] , [17] , [18] , [19] , [20] , other fused-core columns such as C8, HILIC, RP-amide, peptide, phenyl-hexyl are becoming available. In contrast to C18 fused-core columns, little investigation has been carried out on the column performance of these other stationary phases [21] , [22] , [23] . Therefore, we selected the newly available RP-amide stationary phase, for which at this time no quality parameters have been investigated.…”

Section: Resultsmentioning

confidence: 99%

A critical quality parameter in quantitative fused-core chromatography: The injection volume

Boonen

D׳Hondt

Veryser

et al. 2013

Journal of Pharmaceutical Analysis

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As part of the method development, the injection volume as a critical quality attribute in fast fused-core chromatography was evaluated. Spilanthol, a pharmaceutically interesting N-alkylamide currently under investigation in our laboratory, was chosen as the model compound. Spilanthol was dissolved in both PBS and MeOH/H2O (70/30, v/v) and subsequently analyzed using a fused-core system hereby selecting five chromatographic characteristics (retention time, area, height, theoretical plates and symmetry factor) as responses. We demonstrated that the injection volume significantly influenced both the qualitative and quantitative performance of fused-core chromatography, a phenomenon which is confounded with the nature of the used sample solvent. From 2 μL up to 100 μL injection volume with PBS as solvent, the symmetry factor decreased favorably by 20%. Moreover, the theoretical plates and the quantitative parameters (area and height) increased up to 30%. On the contrary, in this injection volume range, the theoretical plates for the methanol-based samples decreased by more than 60%, while the symmetry factor increased and the height decreased, both by 30%. The injection volume is thus a critical and often overlooked parameter in fused-core method description and validation.

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Section: Resultsmentioning

confidence: 99%

A critical quality parameter in quantitative fused-core chromatography: The injection volume

Boonen

D׳Hondt

Veryser

et al. 2013

Journal of Pharmaceutical Analysis

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“…Afterwards, these Cartesian coordinate matrices were used to calculate more than 3000 descriptors, using Dragon 5.5 (Talete, Milan, Italy), HyperChem 8.0 and MarvinSketch 5.10.3 (ChemAxon, Budapest, Hungary) software programs. The specific peptide descriptor LogSum AA , introduced by our research group, was also included in the descriptor set [23]. The non-discriminative descriptors, i.e.…”

Section: Methodsmentioning

confidence: 99%

Chemical-Functional Diversity in Cell-Penetrating Peptides

et al. 2013

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Cell-penetrating peptides (CPPs) are a promising tool to overcome cell membrane barriers. They have already been successfully applied as carriers for several problematic cargoes, like e.g. plasmid DNA and (si)RNA, opening doors for new therapeutics. Although several hundreds of CPPs are already described in the literature, only a few commercial applications of CPPs are currently available. Cellular uptake studies of these peptides suffer from inconsistencies in used techniques and other experimental conditions, leading to uncertainties about their uptake mechanisms and structural properties. To clarify the structural characteristics influencing the cell-penetrating properties of peptides, the chemical-functional space of peptides, already investigated for cellular uptake, was explored. For 186 peptides, a new cell-penetrating (CP)-response was proposed, based upon the scattered quantitative results for cellular influx available in the literature. Principal component analysis (PCA) and a quantitative structure-property relationship study (QSPR), using chemo-molecular descriptors and our newly defined CP-response, learned that besides typical well-known properties of CPPs, i.e. positive charge and amphipathicity, the shape, structure complexity and the 3D-pattern of constituting atoms influence the cellular uptake capacity of peptides.

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“…RP-HPLC can isolate, purify, and prepare antimicrobial peptides based on their unique physicochemical properties and different retention times [17,18]. The active components after purification in Sephadex G50 gel chromatography were freeze dried by vacuum, from which the dried samples were dissolved with methanol and filtered.…”

Section: Anti-phase High-performance Liquid Chromatography (Rp-hplc) ...mentioning

confidence: 99%

Two Antimicrobial Peptides Derived from Bacillus and Their Properties

Zhang,

Meng,

et al. 2023

Molecules

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Growth promotion and disease prevention are important strategies in the modern husbandry industry, and for this reason, antibiotics are widely used as animal feed additives. However, the overuse of antibiotics has led to the serious problem of increasing resistance of pathogenic microorganisms, posing a major threat to the environment and human health. “Limiting antibiotics” and “Banning antibiotics” have become the inevitable trends in the development of the livestock feed industry, so the search for alternative antimicrobial agents has become a top priority. Antimicrobial peptides (AMPs) produced by Bacillus spp. have emerged as a promising alternative to antibiotics, due to their broad-spectrum antimicrobial activity against resistant pathogens. In this study, two strains of Bacillus velezensis 9-1 and B. inaquosorum 76-1 with good antibacterial activity were isolated from commercial feed additives, and the antimicrobial peptides produced by them were purified by ammonium sulfate precipitation, anion exchange chromatography, gel chromatography, and RP-HPLC. Finally, two small molecule peptides, named peptide-I and peptide-II, were obtained from strain 9-1 and 76-1, respectively. The molecular weight and sequences of the peptides were analyzed and identified by LC–MS/MS, which were 988.5706 Da and VFLENVLR, and 1286.6255 Da and FSGSGSGTAFTLR, respectively. The results of an antibacterial activity and stability study showed that the two peptides had good antibacterial activity against Staphylococcus aureus, B. cereus, and Salmonella enterica, and the minimum inhibitory concentrations were 64 μg/mL and 16 μg/mL, 32 μg/mL and 64 μg/mL, and 8 μg/mL and 8 μg/mL, respectively. All of them have good heat, acid, and alkali resistance and protease stability, and can be further developed as feed antibiotic substitutes.

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Reversed-phase fused-core HPLC modeling of peptides

Cited by 14 publications

References 62 publications

A critical quality parameter in quantitative fused-core chromatography: The injection volume

A critical quality parameter in quantitative fused-core chromatography: The injection volume

Chemical-Functional Diversity in Cell-Penetrating Peptides

Two Antimicrobial Peptides Derived from Bacillus and Their Properties

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