Column Chromatography Research Articles

A Streamlined High-Throughput LC-MS Assay for Quantifying Peptide Degradation in Cell Culture.

Peptides are widely used in biomaterials due to their easy of synthesis, ability to signal cells, and modify the properties of biomaterials. A key benefit of using peptides is that they are natural substrates for cell-secreted enzymes, which creates the possibility of utilizing cell-secreted enzymes for tuning cell-material interactions. However, these enzymes can also induce unwanted degradation of bioactive peptides in biomaterials, or in peptide therapies. Liquid chromatography-mass spectrometry (LC-MS) is a widely used, powerful methodology that can separate complex mixtures of molecules and quantify numerous analytes within a single run. There are several challenges in using LC-MS for the multiplexed quantification of cell-induced peptide degradation, including the need for non-degradable internal standards and the identification of optimal sample storage conditions. Another problem is that cell culture media and biological samples typically contain both proteins and lipids that can accumulate on chromatography columns and degrade their performance. However, removing these constituents can be expensive, time consuming, and increases sample variability. Here we show that directly injecting samples onto the LC-MS without any purification enables rapid and accurate quantification of peptide concentration, and that hundreds of LC-MS runs can be done on a single column without a significantly diminish the ability to quantify the degradation of peptide libraries. We also show that column failure is evident when hydrophilic peptides fail to be retained on the column, and this can be easily identified using standard peptide mixtures for column benchmarking. In total, this work introduces a simple and effective method for simultaneously quantifying the degradation of dozens of peptides in cell culture. By providing a streamlined and cost-effective method for the direct quantification of peptide degradation in complex biological samples, this work enables more efficient assessment of peptide stability and functionality, facilitating the development of advanced biomaterials and peptide-based therapies.

A New Megastigmane Nor-sesquiterpene Isolated from Pourouma Guianensis by High-Performance Countercurrent Chromatography

Introduction: Pourouma guianensis (Urticaceae) is widespread throughout Brazil and is commonly known as “mapati”. Terpenes are common in Urticaceae species. However, few studies have reported the isolation and characterisation of these compounds in Pourouma genus. Method: The aim of this study was to separate substances from stem barks of P. guianensis using a high-performance countercurrent chromatography gradient elution method. The dichloromethane extract of the P. guianensis stem bark was submitted to silica gel column chromatography, yielding three fractions. Fraction 3 was submitted to a gradient elution system in high-performance countercurrent chromatography using two solvent systems, hexane/ethyl acetate/methanol/water (1:2:x:1, x=1.0 and 1.5). Six compounds were isolated. Results: A new megastigmane nor-sesquiterpene named 2-acetyl-4,7-epoxy-3-oxy-5,5,9- trimethylhexahydrocoumaran (1) was obtained together with five known terpenes: vomifoliol (2), 2α,3β,19α,23-tetrahydroxyurs-12-en-28-oic acid (3), 3-β-D-arabinopyranoside-19-hydroxyurs-12-en- 28-oic acid (4), arjunolic acid (5), and tormentic acid (6). Fractions 1 and 2 were purified by binary gradient elution from hexane to ethyl acetate. Conclusion: This process isolated three known compounds: friedelin (7) and 3β-friedelanol (8) from fraction 1, and β-sitosterol (9) from fraction 2. Gradient elution in countercurrent chromatography was a suitable alternative, exploiting variations in the partition coefficient between substances with small structural differences.

Anti-Aging Activities and LC-MS Analysis of Chrysophyllum cainito Leaves Extract

Star apple (Chrysophyllum cainito L.), or Apple-nam in Thai, is a medium-sized tree in the Sapotaceae family. It is native to Middle America and distributed in Thailand and Southeast Asia. It has antioxidation, anti-inflammatory and anti-diabetic properties. The active compounds in star apples are flavonoids. This research aimed to study the biological activities and chemical constituents of crude extract and active fractions from the leaves of star apples. The leaf powder was defatted with dichloromethane and further extracted with ethanol using ultrasound-assisted extraction. The ethanol extract was separated by quick column chromatography given 7 fractions (CCE1-CCE7). The total phenolic content and antioxidant activity of the extract and chromatographic fractions were determined. The active fractions were further studied for tyrosinase inhibition, collagenase inhibition and cell proliferation properties. The result showed that the highest total phenolic content was observed in ethanol extract. The high polar fraction, CCE6, displayed the highest antioxidation activity with radical scavenging of 0.34 ± 0.01 mg AAE/mL in the DPPH assay and a FRAP value of 1.36 ± 0.01 mg AAE/mL in the FRAP assay. The CCE6 also exhibited the highest anti-tyrosinase and anti-collagenase activities with a percentage of 67.14 ± 1.34 and 70.32 ± 2.88, respectively and showed the best activity in cell proliferation with a percentage of 24.68 ± 2.94. The analysis of the components using the negative ionization LC-MS technique revealed that gallic acid and quercetin-3-galactoside are the major constituents in the ethanol extract. While the fraction CCE6 showed glucosyl (2E,6E,10x)-10,11-dihydroxy-2,6-farnesadienoate and p-mentha-1,3,5,8-tetraene as the predominant compounds. HIGHLIGHTS The ethanol extract was prepared using ultrasound-assisted extraction. The first report of tyrosinase and collagenase inhibition, as well as cell proliferation properties, of the cainito leaves extract and chromatographic fractions. The major components of the ethanol extract of cainito leaves in Thailand and its active fraction by LC-MS technique were reported for the first time. GRAPHICAL ABSTRACT

Inhibitory Effects of New Epicatechin Oligomers on Nitric Oxide Production.

The primary aim of this research was to identify the structural characteristics of three newly derived procyanidins from cold plasma-treated (-)-epicatechin, known for their anti-inflammatory properties. The newly generated compounds were isolated through column chromatography, and their chemical structures were elucidated through spectroscopic data analyses, including both one-dimensional and two-dimensional nuclear magnetic resonance (NMR) and mass spectrometry (MS) techniques. Furthermore, their absolute configurations were determined via circular dichroism (CD) spectroscopy. The inhibitory activity of the isolated compounds on nitric oxide (NO) production and expression levels of inducible NO synthase (iNOS) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages was evaluated. Three new procyanidins-methylenetrisepicatechin (2), isomethylenetrisepicatechin (3), and methylenebisepicatechin (4)-along with two reported dimeric flavan-3-ols (5 and 6), were identified from plasma-treated (-)-epicatechin (1). The unique oligomerized products 2 and 3 linked by methylene bridges significantly suppressed both NO production and iNOS expression, demonstrating higher anti-inflammatory activities in LPS-stimulated RAW 264.7 cells compared with the parent compound. The newly oligomerized procyanidins have potential applications in the treatment of inflammatory diseases owing to their significant anti-inflammatory properties.

Optimizing Low-Cost Gas Analysis with a 3D Printed Column and MiCS-6814 Sensor for Volatile Compound Detection.

This paper explores an application of 3D printing technology on the food industry. Since its inception in the 1980s, 3D printing has experienced a huge rise in popularity. This study uses cost-effective, flexible, and sustainable components that enable specific features of certain gas chromatography. This study aims to optimize the process of gas detection using a 3D printed separation column and the MiCS-6814 sensor. The principle of the entire device is based on the idea of utilizing a simple capillary chromatographic column manufactured by 3D printing for the separation of samples into components prior to their measurement using inexpensive chemiresistive sensors. An optimization of a system with a 3D printed PLA block containing a capillary, a mixing chamber, and a measuring chamber with a MiCS-6814 sensor was performed. The optimization distributed the sensor output signal in the time domain so that it was possible to distinguish the peak for the two most common alcohols, ethanol and methanol. The paper further describes some optimization types and their possibilities.

Isolation and Identification of Monoterpenoids from Callicarpa Nudiflora.

A phytochemical and directional investigation was carried out on the leaves of Callicarpa nudiflora. Various separation methods including silica gel, sephadex LH-20, medium pressure liquid chromatography (MPLC), preparative thin-layer chromatography (p-TLC) and preparative high performance liquid chromatography (p-HPLC) were used to isolate 10 compounds. The comprehensive structural elucidation of five novel monoterpenoids (1-5) was achieved through multi-spectroscopic analyses, and advanced quantum mechanical electronic circular dichroism (ECD) calculations. Furthermore, the anti-inflammatory activity of the identified compounds was assessed.

Structural characteristics of a purified Evodiae fructus polysaccharide and its gastroprotection and relevant mechanism against alcohol-induced gastric lesions in rats

Evodiae fructus polysaccharide (EFP) has been previously shown to protect against alcohol-induced gastric lesions. However, which and how active fractions in EFP exert gastroprotection remains unclear. This study aimed to characterize the structure of the purified fraction (EFP-2-1) of EFP, and investigate its gastroprotection and underlying mechanisms. EFP-2-1 was obtained through column chromatography, and was characterized using instrumental analytical techniques. Gastroprotective effect of EFP-2-1 was evaluated using alcohol-induced gastric lesions in rats, and its mechanism was explored through proteomics, metabolomics and diversity sequencing. Results showed that EFP-2-1 had a molecular weight of 7.3 kDa, and consisted mainly of rhamnose, galacturonic acid, galactose and arabinose. Its backbone contained HG and RG-I domains, and branched with →5)-α-l-Araf-(1→, α-l-Araf-(1→ and →4)-β-d-Galp-(1→ residues. EFP-2-1 reduced gastric lesions and the levels of MDA, TNF-α and IL-6, activated PPARγ, primarily altered protein digestion and absorption and bile secretion metabolic pathways, regulated gut microbiota like Faecalibaculum and Lachnoclostridium, and increased short-chain fatty acids production. Correlations were observed among the gut microbiota, metabolites and biochemical indexes influenced by EFP-2-1. These findings suggest that EFP-2-1 is an active fraction of EFP for protecting against alcohol-induced gastric lesions, which may be linked to PPARγ activation, gut microbiota and serum metabolism.

Novel Antioxidant Peptides from Fermented Whey Protein by Lactobacillus rhamnosus B2-1: Separation and Identification by in Vitro and in Silico Approaches.

Whey is a byproduct of the dairy industry and is rich in protein. To enhance the significance of such byproducts and find efficacious antioxidants for combating oxidative stress, this study reported on the preparation, purification, and identification of novel peptides with antioxidant activities from whey protein metabolites following fermentation by Lactobacillus rhamnosus B2-1. The isolation and identification processes involved macroporous adsorption resin column chromatography, gel filtration column chromatography, and liquid chromatography-tandem mass spectrometry. Therein, three novel antioxidant peptides (PKYPVEPF, LEASPEVI, and YPFPGPIHNS) were selected to be synthesized, and they demonstrated remarkable antioxidant activities in vitro chemical assays. PKYPVEPF, LEASPEVI, and YPFPGPIHNS (100 μg/mL) displayed a notable cytoprotective impact on HepG2 cells under oxidative stress induced by H2O2, increasing the cell viability from 49.02 ± 3.05% to 88.59 ± 10.49%, 82.38 ± 19.16%, and 85.15 ± 7.19%, respectively. Moreover, the peptides boosted the activities of catalase and superoxide dismutase in damaged cells and reduced reactive oxygen species levels. The molecular docking studies highlighted that these antioxidant peptides efficiently bound to key amino acids in the Kelch domain of Keap1, thereby preventing the interaction between Keap1 and Nrf2. In conclusion, PKYPVEPF, LEASPEVI, and YPFPGPIHNS demonstrated substantial antioxidant activity, suggesting their potential for widespread application as functional food additives and ingredients.

HPLC analysis and antimicrobial activity of Vitex altissima leaves extracts

The exploration of ethnomedicinal plants for phytochemical and pharmacological properties is crucial for developing novel therapeutics for chronic diseases. Vitex altissima, known as the Peacock chaste tree, is a significant member of the Verbenaceae family, is widely utilized in traditional medicine. This study investigates the phytochemical composition and antibacterial properties of V. altissima, a plant recognized for its medicinal use by various indigenous communities, including the Malayali tribes of Servarayan hills. The study involved extracting phytochemicals from the leaves of V. altissima using hexane, ethyl acetate and methanol through soxhlet extraction. The extracts were analyzed for secondary metabolites, including carbohydrates, proteins, amino acids, steroids, glycosides, alkaloids, tannins, phenolics, flavonoids, terpenoids, saponins, anthraquinones, oils and resins, diterpenes, phlobatannins and coumarins. Quantitative assessments of phenolic compounds, flavonoids and alkaloids were performed. Column chromatography was employed to fractionate the methanolic extract, resulting in 12 distinct fractions. These fractions were screened for antibacterial activity against E. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus mutans and Staphylococcus aureus using the agar well diffusion method. The minimal inhibitory concentration (MIC) was determined through a 2-fold serial dilution technique. High-performance liquid chromatography (HPLC) was utilized to identify the polyphenolic compounds responsible for antibacterial activity. Phytochemical screening revealed a diverse array of bioactive compounds in the extracts, with methanol extracts showing the highest total phenolic content (218 ± 7.21 mg GAE/g of extract) and significant antibacterial activity. The ethyl acetate extract exhibited the highest total flavonoid content (40.67 ± 6.65 mg QE/g of extract). Among the fractions, the column fraction from the chloroform-methanol gradient (VACF) demonstrated superior antimicrobial activity, particularly against S. mutans and P. aureginosa. The MIC values for VACF were 427 µg/mL for P. aeruginosa and 400 µg/mL for S. aureus, indicating potent antibacterial properties. HPLC analysis identified key polyphenolic compounds, including p-coumaric acid, ferulic acid and elagic acid, as the primary contributors to the antibacterial activity. The presence of these compounds aligns with the observed antimicrobial efficacy and highlights the potential of V. altissima extracts as a source of natural antibacterial agents.

Qualitative and Quantitative Sex-Related Differences in the Perception of Single Molecules from Coffee Headspace.

One of the still-debated topics regarding the olfactory function concerns the presence or absence of sex-related differences in individuals. In this study, we checked for a relationship between the olfactory function of females and males and their ability to perceive single molecules, and researched how this can influence the intensity with which the complex odor formed by a set of single molecules is perceived. First, females and males were classified as normosmic or hyposmic based on the TDI olfactory score obtained using the Sniffin' Sticks test. Subsequently, the headspace of roasted coffee beans, as a complex olfactory stimulus, was broken down into single molecules by means of a chromatographic column; these were simultaneously conveyed to a mass spectrometer (for their subsequent classification) and to the human nose, which acts as a chemical sensor by means of an olfactometer port. The results obtained with this gas chromatography-olfactometry approach show both qualitative and quantitative differences between females and males, with females performing better than males. In addition, the odor intensity reported by females when sniffing pen #10, containing coffee aroma, is significantly higher than that reported by males. In conclusion, these data highlight that the human ability to perceive both single compounds and complex odors is strongly conditioned, not only by the olfactory function of individuals, but also by their sex.

Revealing the backbone structures of land-based fulvic acids derived from river and soil through n-butylsilane reduction.

While ultra-high-resolution mass spectrometry has enabled the identification of the molecular composition of dissolved organic matter (DOM), elucidating its molecular structure remains a challenging endeavor. Here, two fulvic acids (FAs), one from river and the other from forest soil, were subjected to reduction using an optimized n-butylsilane (n-BS) reduction method. The reduction products were purified through a combination of liquid-liquid extraction and silica gel column chromatography, resulting in the separation into saturates, aromatics, and polar products. The polar products were analyzed by high-resolution mass spectrometry (HRMS), and the saturates and aromatics were analyzed using gas chromatography-mass spectrometry (GC-MS). HRMS results showed that the number of oxygen atoms and double-bond equivalent (DBE) values of FA decreased after reduction. GC-MS results revealed that a total of 270 hydrocarbon monomers were identified from the reduction products of a single sample, with the highest carbon number of cycloalkanes reaching C33. For the first time, steranes and hopanes were detected in the reduction products, potentially serving as evidence for the existence of carboxyl-rich alicyclic molecule (CRAM) precursors. Additionally, a significant number of polycyclic aromatic hydrocarbons were identified, and the potential sources of various compounds were preliminarily inferred based on their isomers. This study extends the knowledge of the possible backbone structure of the DOM and provides a new potential tool for investigating the origin and transformation mechanisms of DOM.

Identification and in vitro genotoxicity assessment of forced degradation products of glimepiride and glyburide using HEK cell-based COMET assay.

This study focuses on characterizing the forced degradation products of antidiabetic drugs glimepiride (GMD) and glyburide (GBD), with previously unexplored genotoxicity. Drugs underwent stress induced by acid, base, and hydrogen peroxide. For GMD, impurities were profiled and isolated using Hypersil Gold C8 (250 × 10 mm, 5μ) through semi-preparative HPLC with a fraction collector. For GBD, impurity profiling was performed using semi-preparative HPLC (Hypersil GOLD C18, 250 × 10 mm, 5μ), and reverse-phase flash chromatography (FP ECOFLEX C18 4g column) for isolation. Although five GMD and three GBD impurities were detected, only three GMD and two GBD impurities were separated and assessed for purity using analytical RP-HPLC with the purity percentages ranging from 96.6% to 99.9%. LC-Orbitrap MS was used to identify these three GMD impurities (m/z: 408.122, 338.340, 381.160) and two GBD impurities (m/z: 369.065, 325.283). ProTox-II in silico predictions classified all impurities as class 4 and 5, with no positive genotoxicity indications. In vitro comet assays, using HEK cells, indicated that for GMD, impurity 2 and impurity 5 were less genotoxic, whereas impurity 4 exhibited genotoxicity. For GBD, both impurities 1 and 3 were found to be genotoxic, with impurity 3 showing a higher level of genotoxicity than impurity 1.

Structural characterization of a cell wall-associated polysaccharide of Nocardia rubra FIM-21 and its immunostimulatory activity

In this work, a neutral polysaccharide named NP-1 was obtained from the cell wall of Nocardia rubra FIM-21 by column chromatography. Its chemical structure was characterized by methylation analysis, 2D NMR spectroscopic study, and other techniques. It was shown that NP-1 contained five types of monosacchrides, including mannose (Man), glucose (Glc), N-acetylglucosamine (GlcpNAc), galactose (Gal) and arabinose (Ara) in a molar ratio of 7.7:3:2:1:1.5. According to linkage analysis and NMR spectra, the backbone contains 1,6-linked Manp, 1,4-linked Glcp, and 1,6-linked Galp, with branches substituted at the C-6 of 1,4-linked Glcp and the C-2 of 1,6-linked Galp by GlcpNAc, and at C-2 of 1,6-linked Manp by Araf, Araf-1,5-Araf or GlcpNAc. Biological activity research showed that NP-1 could stimulate the proliferation of RAW264.7 cells and enhance their phagocytosis and release of NO.

Sustainable methods for producing food-derived bioactive compounds

Background: Pursuing alternative starting materials has gained significant importance in the food and drug industries, driven by the global shift towards sustainable and renewable resources. Terpenes, naturally occurring hydrocarbons derived from plants, present a promising solution due to their abundant availability and range of physiological activities, including antioxidant, anti-inflammatory, antimicrobial, and anticancer properties. Objective: This study aims to develop a waste-free synthesis methodology for functionalizing limonene and myrcene, two notable terpenes, through Atom Transfer Radical Reactions (ATRR). The objective is to achieve high yields and purities while enhancing the therapeutic potential of these compounds and adhering to green chemistry principles. Methods: A catalytic complex of Cu(I)Br and dimethyl sulfoxide (DMSO) was used to facilitate the coupling of limonene and myrcene with various trichloroacetic acid derivatives. Reactions were performed at 80°C for 90 minutes, and products were purified by column chromatography. The synthesized compounds were characterized using Gas Chromatography-Mass Spectrometry (GC-MS) and Nuclear Magnetic Resonance (NMR). Microbiological analyses were conducted using the disk-diffusion method, and statistical analyses were performed to assess the reliability of the experiments. Results: The ATRR methodology enabled the efficient functionalization of limonene and myrcene, achieving high yields and purities. The best yield for limonene was 70%, and for myrcene, 82%, both using trichloroacetonitrile. The Cu(I) catalytic system showed dualistic biological effects, with 3% concentration exhibiting strong bactericidal and fungicidal activity, while 0.03% promoted growth. These results demonstrate the potential of the methodology and the catalytic system for bioactive compound synthesis and agricultural applications. Conclusion: This study demonstrates the viability of ATRR for efficiently functionalizing limonene and myrcene, achieving high yields and purities. The synthesized compounds show promise for pharmaceutical development. The dual antibacterial and antifungal activities of the Cu(I) catalytic system also suggest potential applications in wastewater reutilization. Future work should explore the biological effects and environmental impact of this process. Keywords: Terpenes, ATR reactions, Green Chemistry, Bioactive compounds, Therapeutic potential.

Dopamine-supported HPLC post-column derivatization to fluorescence: Simultaneous and sensitive detection of eight tea polyphenols

The effective differentiation and detection of multiple tea polyphenols are often challenging due to their subtle structural similarities. Although post-column derivatization HPLC strategies are commonly employed to distinguish multiple targets, the short physical distance between chromatographic column and detector limits reaction time, thereby reducing the derivatization efficiency. Dopamine (DA) reacts rapidly with resorcinol to form fluorescent azamonardine products, making fast fluorometric derivatization of tea polyphenols containing resorcinol motifs possible. In this study, a DA-driven rapid and post-column fluorescence derivatization method has been applied to sensitively detect eight tea polyphenols. This method is based on fluorescence derivatization and possesses low background interference, high sensitivity, and excellent reproducibility. Moreover, the practical application of this proposed fluorometric derivatization platform was further validated by simultaneous identification of multiple tea polyphenols in different tea samples. This work has great potential to become an alternative to the National Standard method for tea polyphenols determination.

Introducing Column Chromatography through Colorful Reactions of Guaiazulene

Introducing Column Chromatography through Colorful Reactions of Guaiazulene

Consolidated Curium Reprocessing Procedure Inspires Molecular Design for Sensitized Curium Circularly Polarized Luminescence.

Curium's stable redox chemistry and ability to emit strong metal-based luminescence make it uniquely suitable for spectroscopic studies among the actinide series. Targeted ligand and coordination compound design can support both fundamental electronic structure studies and industrial safeguards with the identification of unique spectroscopic signatures. However, limited availability, inherent radioactive hazards, and arduous purifications have long inhibited such investigations of this element. A consolidated reprocessing procedure for curium has been developed for the milligram scale. The recovery of not only standard legacy curium samples but also hazardous legacy perchlorate containing curium samples was achieved, culminating in column chromatography utilizing the extraction resin DGA (N,N,N',N'-tetra-2-ethylhexyldiglycolamide, branched). Surprisingly, controlled elution of the Cm band from the extraction resin was followed through bright pink luminescence triggered by an inexpensive hand-held UV-vis lamp (380-400 nm). This observation inspired the design of an enantiopure, C2-symmetrical ligand bearing a chiral (trans-1,2-diaminocyclohexane) backbone with achiral DGA moieties (N,N,N',N'-tetra-n-octylacetamide), that enabled rarely observed curium circularly polarized luminescence upon metal chelation. These combined achievements should unlock more luminescence and circularly polarized luminescence studies of curium, and enable the recovery of many curium and other trivalent actinide samples.

Encapsulation of Phloroglucinol from Rosenvingea intricata Macroalgae with Zinc Oxide Nanoparticles against A549 Lung Cancer Cells

Background/Objectives: Phloroglucinol (PHL), a phenolic compound extracted from the brown alga Rosenvingea intricata, exhibits potent antioxidant and anticancer properties. This study aims to extract, purify, and characterize PHL, and further develop functionalized zinc oxide nanoparticles (ZnO NPs) loaded with PHL to enhance its therapeutic potential. Methods: PHL was extracted using acetone and purified through Sephadex LH-20 column chromatography, yielding a highly enriched fraction (F-3). The purified compound was characterized by FTIR, HPLC, NMR, and LC-MS. ZnO NPs were synthesized, PEGylated, and conjugated with PHL, forming ZnO-PEG-PHL NPs. Their characterization included DLS, zeta potential, XRD, SEM-EDAX, and encapsulation efficiency studies. Antioxidant assays (DPPH, FRAP, ABTS, RPA) were performed and in vitro cytotoxicity on A549 lung cancer cells were determined to evaluate the therapeutic efficacy of PHL. Results: The purified PHL fraction showed a high phenolic content (45.65 PHL mg/g), which was was confirmed by spectral analysis. The ZnO-PEG-PHL NPs increased in size from 32.36 nm to 46.68 nm, with their zeta potential shifting from −37.87 mV to −26.82 mV. The antioxidant activity was superior for the ZnO-PEG-PHL NPs in all assays, while the in vitro cytotoxicity tests showed an IC50 of 40 µg/mL compared to 60 µg/mL for the ZnO NPs and 70 µg/mL for PHL. Apoptotic studies revealed significant cell cycle arrest and apoptosis induction. Conclusions: The synthesized ZnO-PEG-PHL NPs demonstrated enhanced antioxidant and anticancer properties, making them promising candidates for cancer therapy and antioxidant applications.

Microchip gas chromatographic columns dedicated for space exploration: Stationary phase coating, setup optimization and evaluation of column performances

This work is the first part of a project aiming to specifically design gas chromatographic microcolumns for space exploration. In particular, this study explored the functionalization and characterization of silicon/glass microchip gas chromatographic columns designed with a serpentine-shaped channel having an internal square cross-section. This microcolumn can be connected using a robust fluidic manifold with removable capillary connections to a conventional gas chromatograph or implemented in a prototype instrument for space exploration. First, benchtop gas chromatographic system was optimized in terms of injector liner volume, internal diameter (I.D.) of the capillary connections and data frequency of detection to ensure an optimal evaluation of column efficiency. Junction capillaries of 100 μm I.D., a liner of 1.2 mm I.D. and a detection acquisition frequency of 100 Hz were found to be the optimal set-up and parameters. Moreover, the stationary phase coating velocity was slowed-down to increase column efficiency by 100 %. Then, the performances of a square cross-section capillary column were compared with those of a conventional circular cross-section column. The coating efficiencies were estimated at 55 % and 42 % for circular and square internal cross-section geometries respectively, demonstrating that the square section geometry did not affect significantly the column performances. Stationary phase films of different thicknesses, from 0.032 to 0.260 µm, were coated on different microchips of the same production batch to assess the influence of film thickness on the chromatographic performances. Microchips performance was evaluated through Golay's plots, and some of the microchips were also studied by optical microscopy. The efficiency and retention capacity of our microchip column is shown to be highly dependent on the film thickness (9,000 plates.m-1 with k = 0.12 for the thinnest film and up to almost 4,000 plates.m-1 with k = 0.93 for the thickest film). Finally, the coating process repeatability was validated by producing three identical microchips with RSD of 4.8 % for the average number of theoretical plates.

A readily accessible quaternized cellulose filter paper with high permeability for IgG separation

Anion-exchange chromatography (AEC) is recognized as a highly effective approach for the purification of immunoglobulin G (IgG). This study introduces an innovative strategy that employs waste cellulose filter paper in the production of AEC media. A quaternized cellulose fiber membrane (CFM-QCS) was successfully fabricated that cellulose fibers were as a structural framework, glutaraldehyde (GA) as a crosslinking agent, and quaternary chitosan (QCS) as a modifying agent. Morphological and chemical characterization revealed that GA and QCS were uniformly crosslinked on the surface of the cellulose fibers, resulting in excellent mechanical properties in both dry and wet states. Benefiting from its 3D network scaffold structure, CFM-QCS demonstrated a high adsorption capacity for bovine serum albumin (BSA), with static and dynamic adsorption capacities of 605.15 mg/g and 88.63 mg/ml, respectively. After treated with extreme conditions and 10 cyclic adsorption and elution, the adsorption capacity of CFM-QCS remains almost unchanged, highlighting its excellent stability. Additionally, a CFM-QCS packed chromatography column exhibited high flux of 10.38 L/h at 0.1 MPa, which can efficiently separate IgG from a mixed solution in the presence of BSA and IgG by gravity-driven. This work presents a straightforward approach for preparing high-performance ion-exchange chromatography membranes for IgG separation.