Determination of Glycoalcaloids in Hungarian Potatoes by HPLC

In this work potato genotypes originated from an intensive resistance breeding programme that includes unique combination of resistance to several pathogens and consumer response were analysed for glycoalcaloid content by reversed phase high performance liquid chromatographic method. The glycoalcaloids were concentrated from potato samples by solid-phase extraction with a disposable C18 cartridge column. Nucleosil 100-5 C-18-AB column was used with 50% acetonitrile in phosphate buffer as the mobile phase. The method was validated according to calibration curve, precision and recovery tests, limit of detection and quantitation. The results indicated that as low as 0.27 µg mL⁻¹ can be quantitatively determined by this method with 94–98% and 99% for recovery and precision, respectively. Hungarian potatoes were analysed by the adapted procedure for their glycoalcaloid content. The result showed that the tested Hungarian potato varieties contain glycoalcaloids between 0.09 and 15 mg 100 g⁻¹. The content of glycoalcaloids in the new varieties of potato was lower than the limit of the Official Food Regulations.

     

Simultaneous determination of Sudan dyes and carotenoids in red pepper and tomato products by HPLC

To simultaneously detect Sudan dyes and carotenoids in red pepper and tomato products, high-performance liquid chromatography (HPLC) methods with photodiode-array detection are developed and validated. The methods include the use of end-capped and nonend-capped adsorbents with a gradient elution system starting with water containing methanol. Water content of 9% in the starting mobile phase is found to be necessary to ensure sufficient separation of Sudan dyes and to avoid overlapping or interference with the carotenoids of considerable content. The data of the validation reveal the accuracy and precision of the developed methods. A limit of Sudan dyes detection of 1-5 microg/g in red pepper or tomato sauce could be approached. The methods provide excellent separation of the carotenoids from the unsaponified extracts of red pepper and the tomato products.     

Liquid Chromatographic Analysis of Phenolic Compounds in Organically and Conventionally Grown Varieties of Sour Cherries

Liquid chromatographic analysis of the polyphenols in sour cherries was achieved on bi-functional RP columns with a gradient prepared from methanol and aqueous formic acid. For quantitative analysis the compounds were detected at 320 nm. The method was validated for linearity, precision, and recovery. Chlorogenic acid, neochlorogenic acid, and rutin were detected in sour cherry cultivars. Amounts of neochlorogenic acid were in the range 471–590 μg g⁻¹. For the three sour cherry cultivars investigated in this study there was no significant difference between the chlorogenic acid and rutin content of cherries produced by conventional and organic farming. There were, however, significant differences between the amounts of these compounds in cherries harvested in different years.

     

Molecular Dynamic Simulation Search for Possible Amphiphilic Drug Discovery for Covid-19

To determine whether quaternary ammonium (k21) binds to Severe Acute Respiratory Syndrome–Coronavirus 2 (SARS-CoV-2) spike protein via computational molecular docking simulations, the crystal structure of the SARS-CoV-2 spike receptor-binding domain complexed with ACE-2 (PDB ID: 6LZG) was downloaded from RCSB PD and prepared using Schrodinger 2019-4. The entry of SARS-CoV-2 inside humans is through lung tissues with a pH of 7.38–7.42. A two-dimensional structure of k-21 was drawn using the 2D-sketcher of Maestro 12.2 and trimmed of C18 alkyl chains from all four arms with the assumption that the core moiety k-21 was without C18. The immunogenic potential of k21/QA was conducted using the C-ImmSim server for a position-specific scoring matrix analyzing the human host immune system response. Therapeutic probability was shown using prediction models with negative and positive control drugs. Negative scores show that the binding of a quaternary ammonium compound with the spike protein’s binding site is favorable. The drug molecule has a large Root Mean Square Deviation fluctuation due to the less complex geometry of the drug molecule, which is suggestive of a profound impact on the regular geometry of a viral protein. There is high concentration of Immunoglobulin M/Immunoglobulin G, which is concomitant of virus reduction. The proposed drug formulation based on quaternary ammonium to characterize affinity to the SARS-CoV-2 spike protein using simulation and computational immunological methods has shown promising findings.     

Concentration-Dependent Multi-Potentiality of L-Arginine: Antimicrobial Effect,Hydroxyapatite Stability,and MMPs Inhibition

This study’s objective was to examine L-arginine (L-arg) supplementation’s effect on mono-species biofilm (Streptococcus mutans/Streptococcus sanguinis) growth and underlying enamel substrates. The experimental groups were 1%, 2%, and 4% arg, and 0.9% NaCl was used as the vehicle control. Sterilised enamel blocks were subjected to 7-day treatment with test solutions and S. mutans/S. sanguinis inoculum in BHI. Post-treatment, the treated biofilms stained for live/dead bacterial cells were analysed using confocal microscopy. The enamel specimens were analysed using X-ray diffraction crystallography (XRD), Raman spectroscopy (RS), and transmission electron microscopy (TEM). The molecular interactions between arg and MMP-2/MMP-9 were determined by computational molecular docking and MMP assays. With increasing arg concentrations, bacterial survival significantly decreased (p < 0.05). The XRD peak intensity with 1%/2% arg was significantly higher than with 4% arg and the control (p < 0.05). The bands associated with the mineral phase by RS were significantly accentuated in the 1%/2% arg specimens compared to in other groups (p < 0.05). The TEM analysis revealed that 4% arg exhibited an ill-defined shape of enamel crystals. Docking of arg molecules to MMPs appears feasible, with arg inhibiting MMP-2/MMP-9 (p < 0.05). L-arginine supplementation has an antimicrobial effect on mono-species biofilm. L-arginine treatment at lower (1%/2%) concentrations exhibits enamel hydroxyapatite stability, while the molecule has the potential to inhibit MMP-2/MMP-9.     

Simple in-line postcolumn oxidation and derivatization for the simultaneous analysis of ascorbic and dehydroascorbic acids in foods

A new analytical procedure for the simultaneous determination of L-ascorbic acid (AA), isoascorbic acid (IAA), L-dehydroascorbic acid (DHAA), and isodehydroascorbic acid (IDHAA) in food by high-performance liquid chromatography (HPLC) is developed. After separation on an HPLC column, an in-line oxidation of AA and IAA to DHAA and IDHAA, respectively, is performed on a short column of activated charcoal. The dehydroascorbic acids are derivatized with a 1,2-phenylenediamine solution in a heated capillary Tefzel reactor into fluorescent quinoxaline compounds and monitored fluorometrically. The chromatographic method provides good separation of LAA, LDHAA, and their diastereoisomers in a relatively short time (-10 min). After optimization of postcolumn derivatization conditions, calibration runs and recovery tests are performed. The fluorescent response in terms of peak area is highly proportional to the concentration of all derivatives examined over a range of 0.1 to 100 microg/mL solution for LAA, LDHAA, IAA, and IDHAA. Recoveries were in the range of 97 to 103%. The detection limit is 0.1 mg of each ascorbic acid derivative per 100 g food. A wide variety of foods (fruits, fruit juices, vegetables, vegetable products, milk, liver, and sausage) are analyzed by the developed procedure. The Vitamin C (LAA and LDHA) contents determined according to the present analytical method are in the same order of magnitude as the result of precolumn derivatization and the fluorometric methods. The described method is a highly specific procedure for determining Vitamin C in food. It is simple to handle, only slightly susceptible to disturbance, perfectly suitable for serial determinations, and yields reproducible results.     

The simultaneous determination of capsaicinoids,tocopherols, and carotenoids in pungent pepper powder

From nutritional points of view, carotenoids, capsaicinoids, and tocopherols are valuable constituents in pungent peppers. A rapid and reliable high performance liquid chromatography (HPLC) method for the simultaneous determination of phytonutrients in spice red peppers and chili products was developed and validated. The method included simultaneous detection by fluorescence and diode-array detectors. The major capsaicinoids, two tocopherols and 43 carotenoid components, were simultaneously separated, detected, and identified in the appointed pepper powder (containing Capsicum annuum and Capsicum frutescens) for method validation. The separation was performed on a Nucleosil C18 reverse phase column and optimized gradient elution. Resolution ranged between 0.96 and 1.46 with the highest values corresponding to γ-tocopherol and α-tocopherol. The limits of detection and quantification of target compounds ranged between 18.77 and 148.08 ng mL^?1. Recoveries were between 89.83–100.26 and 79.72–88.86% when standard materials were spiked at low and high amounts, respectively. The most sufficient extraction of the different phytonutrients was achieved by mixture of methanol and acetone, although it was only slightly better than the mixture of methanol and acetonitrile. These results suggest that the developed method could be used for rapid, one-step determination of a wide range of phytonutrients in chili and pepper powders.