High-performance Chromatography Research Articles

In vitro combination effects of plant-derived quercetin with synthetic bicalutamide on prostate cancer and normal cell lines: in silico comparison.

Prostate cancer is the second most frequent and the fifth greatest cause of death in men. Although diet has been connected to the prevalence of cancer in addition to other factors, the relation between cancer and prevention is weak. Treatment options are at risk due to cell resistance. To identify new combinations, we tried plant-derived quercetin with bicalutamide on cell lines. To determine the cytotoxicity and apoptotic potential of plant-derived quercetin and its combination, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide] and dual stain assays were performed. In silico protein-ligand interaction was performed to support the in vitro findings. A thin layer, column, and high-performance chromatography were used to purify quercetin along with an authentic sample. In the cytotoxic study, quercetin was minimized by 80% similar to bicalutamide and a combination of quercetin and bicalutamide by 50% when compared to controls by 2%. Quercetin and bicalutamide showed a similar binding affinity for androgen receptors (9.7 and 9.8), hub genes (10.8 and 10.0), and a few other PCa-related genes (9.4 and 9.1). We propose to conclude that the combination of quercetin plus bicalutamide can be used for chemotherapy if additional in vivo studies are conducted. The intake of foods high in polyphenolic compounds can help to prevent prostate cancer. Examination of quercetin on several cell lines will provide a definite conclusion to combat cancers.

Aflatoxin profiles of Aspergillus flavus isolates in Sudanese fungal rhinosinusitis.

Aspergillus flavus is a commonly encountered pathogen responsible for fungal rhinosinusitis (FRS) in arid regions. The species is known to produce aflatoxins, posing a significant risk to human health. This study aimed to investigate the aflatoxin profiles of A. flavus isolates causing FRS in Sudan. A total of 93 clinical and 34 environmental A. flavus isolates were studied. Aflatoxin profiles were evaluated by phenotypic (thin-layer and high-performance chromatography) and genotypic methods at various temperatures and substrates. Gene expression of aflD and aflR was also analyzed. A total of 42/93 (45%) isolates were positive for aflatoxin B1 and AFB2 by HPLC. When the incubation temperature changed from 28°C to 36°C, the number of positive isolates decreased to 41% (38/93). Genetic analysis revealed that 85% (79/93) of clinical isolates possessed all seven aflatoxin biosynthesis-associated genes, while 27% (14/51) of non-producing isolates lacked specific genes (aflD/aflR/aflS). Mutations were observed in aflS and aflR genes across both aflatoxin-producers and non-producers. Gene expression of aflD and aflR showed the highest expression between the 4th and 6th days of incubation on the Sabouraud medium and on the 9th day of incubation on the RPMI (Roswell Park Memorial Institute) medium. Aspergillus flavus clinical isolates demonstrated aflatoxigenic capabilities, influenced by incubation temperature and substrate. Dynamic aflD and aflR gene expression patterns over time enriched our understanding of aflatoxin production regulation. The overall findings underscored the health risks of Sudanese patients infected by this species, emphasizing the importance of monitoring aflatoxin exposure.

Novel Stable Protease Inhibitor from Phoenix dactylifera(L.) Flowers with Antimicrobial and Antitumoral Activities.

A novel protease inhibitor isolated from date palm Phoenix dactylifera(L.) flowers (PIDF) was purified and characterized. A heat and acidic treatment step followed by ethanol precipitation and reverse-phase high-performance chromatography was applied to purify this natural protease inhibitor to homogeneity with a single band of about 19 kDa. The stability study depicted that PIDF was fully stable at 40 °C and retained 65% of its initial activity after heating at 50 °C for 24 h. Its thermal stability at 70 °C was markedly enhanced by adding calcium, bovine serum albumin, and sorbitol as well as by metal divalent cations, especially Mg2+ and Hg2+. This protease inhibitor showed high inhibitory activity against therapeutic proteases, including pepsin, trypsin, chymotrypsin, and collagenase, and acted as a potent inhibitor of some commercial microbial proteases from Aspergillus oryzae, Bacillus. sp, and Bacillus licheniformis. Moreover, a potent antibacterial spectrum against Gram (+) and Gram (-) bacterial strains and an efficient antifungal effect were observed. Its cytotoxicity toward human colorectal cancer cell LoVo and HCT-116 lines suggested that PIDF could serve as a new therapeutic target inhibiting human colorectal cancer.

Oral administration of a recombinant modified RBD antigen of SARS-CoV-2 as a possible immunostimulant for the care of COVID-19

BackgroundDeveloping effective vaccines against SARS-CoV-2 that consider manufacturing limitations, equitable access, and acceptance is necessary for developing platforms to produce antigens that can be efficiently presented for generating neutralizing antibodies and as a model for new vaccines.ResultsThis work presents the development of an applicable technology through the oral administration of the SARS-CoV-2 RBD antigen fused with a peptide to improve its antigenic presentation. We focused on the development and production of the recombinant receptor binding domain (RBD) produced in E. coli modified with the addition of amino acids extension designed to improve antigen presentation. The production was carried out in shake flask and bioreactor cultures, obtaining around 200 mg/L of the antigen. The peptide-fused RBD and peptide-free RBD proteins were characterized and compared using SDS-PAGE gel, high-performance chromatography, and circular dichroism. The peptide-fused RBD was formulated in an oil-in-water emulsion for oral mice immunization. The peptide-fused RBD, compared to RBD, induced robust IgG production in mice, capable of recognizing the recombinant RBD in Enzyme-linked immunosorbent assays. In addition, the peptide-fused RBD generated neutralizing antibodies in the sera of the dosed mice. The formulation showed no reactive episodes and no changes in temperature or vomiting.ConclusionsOur study demonstrated the effectiveness of the designed peptide added to the RBD to improve antigen immunostimulation by oral administration.Graphical

Phytochemical Analysis, acetylcholinesterase inhibition, antidiabetic and antioxidant activity of Atriplexhalimus L. (Amaranthaceae Juss.).

Mediterranean saltbush AtriplexhalimusL. (Aranthaceae) from different bioclimatic arid zones (ten wild populations) were studied. Phenols contents, flavonoids, flavonols, tannins and anthocyanins were determined and then tested for their antioxidants, antidiabetic and anti-acetylcholinesterase (AChE)activities. Levels of total polyphenols including flavonoids and flavonols, Tannins and anthocyanins were high and varied significantly among analyzed populations.Nine phenolic acids and four flavonoids were identified for the first time in the methanolic fraction and quantified byliquid high-performance chromatography system HPLC (DAD). All extracts showed a substantial antioxidant activity, as assessed by DPPH assay (1,1-diphenyl-2-picrylhydrazyl free radical) (IC50DPPH=147.3for population of Seliena), Ferric Reducing Antioxidant Power (FRAP; IC50FRAP=3.2 for populations of Sousse and Kairouan), and Chelation Fer test (IC50FerCh=1.5 µg/mL for populations of El-hamma and Mednine).Atriplex halimuspossessed a high inhibitory effect against α-amylase activity(up to 2.6 mg ACE/gE), a moderate activity for α-glucosidase (up to 91.0 mg ACE/gE) and AChE (up to 147.2 µg/mL) compared to standard. The analyzed populations were isolated and subdivided into three distinct groups, without any bioclimatic structuration. Enzymatic activities seem to be associated with the presence, in extracts, of other classes of compounds then phenols such as terpenes,sterols, saponins, coumarins and carotenoids.

Simple Routes to Stable Isotope-Coded Native Glycans.

Understanding the biological role of protein-linked glycans requires the reliable identification of glycans. Isomer separation and characterization often entail mass spectrometric detection preceded by high-performance chromatography on porous graphitic carbon. To this end, stable isotope-labeled glycans have emerged as powerful tools for retention time normalization. Hitherto, such standards were obtained by chemoenzymatic or purely enzymatic methods, which introduce, e.g., 13C-containing N-acetyl groups or galactose into native glycans. Glycan release with anhydrous hydrazine opens another route for heavy isotope introduction via concomitant de-N-acetylation. Here, we describe that de-N-acetylation can also be achieved with hydrazine hydrate, which is a more affordable and less hazardous reagent. Despite the slower reaction rate, complete conversion is achievable in 72 h at 100 °C for glycans with biantennary glycans with or without sialic acids. Shorter incubation times allow for the isolation of intermediate products with a defined degree of free amino groups, facilitating introduction of different numbers of heavy isotopes. Mass encoded glycans obtained by this versatile approach can serve a broad range of applications, e.g., as internal standards for isomer-specific studies of N-glycans, O-glycans, and human milk oligosaccharide by LC-MS on either porous graphitic carbon or─following permethylation─on reversed phase.

Determination of Aflatoxin M1 in raw cow milk, raw camel milk, and heat-processed milk in the North region of Iran

ABSTRACT In this study, 40 milk samples (10 raw cow milk, 10 camel milk, 10 pasteurised milk, and 10 ultra-high temperature milk) were randomly collected from Golestan Province (including two cities, Gorgan and Gonbad-e Kavus) from December 2021 to March 2022. Aflatoxin M1(AFM1) level was determined by the high-performance chromatography method. AFM1 was found in all samples. The mean concentration of AFM1 in raw cow milk, camel milk, pasteurised milk, and ultra-high temperature milk were 72.81 ± 2.18, 57.10 ± 1.86, 34.73 ± 1.34, and 49.36 ± 1.78 ng/kg, respectively. The highest level of AFM1 was found in raw cow milk (p < 0.05). According to the EU standard of AFM1 level, pasteurised milk is the safest type of milk in Golestan Province.

Spent coffee grounds as a potential culture medium for γ-aminobutyric acid (GABA) production by Levilactobacillus brevis PML1

This study aimed to investigate the Gamma-aminobutyric acid (GABA) production potential of Levilactobacillus brevis PML1 using an industrial waste-based culture medium. The optimization of GABA production was carried out in Spent Coffee Grounds (SCG) medium employing Response Surface Methodology (RSM). A Central Composite Design (CCD) was employed, revealing that the optimal SCG treatment conditions were as follows: 6% H2SO4 concentration, 120 min incubation, 0.14 mL Accellerase-1000 enzyme. High-performance chromatography analysis demonstrated a GABA yield of 492 mg/L after 48 h at 37 °C when supplemented with 3% monosodium glutamate. The antimicrobial activity assessment exhibited sensitivity towards Staphylococcus aureus among tested pathogenic bacteria. Scanning electron microscopy elucidated the antibacterial mechanism on Escherichia coli and S. aureus as representative strains. Furthermore, radical inhibitory activities were observed through DPPH assay recording a maximum inhibition rate of 13.15%, while FRAP assay indicated an antioxidant capacity of up to 0.419 mmol/L. Also, L. brevis demonstrated 53.40% cholesterol removal capability. Cytotoxic effects on human colon cancer cell lines Caco-2 were examined using MTT assay, yielding an IC50 value of approximately 3.25 g/mL for the extracts. In conclusion, this study highlights that SCG can serve as an affordable culture medium for efficient metabolite production.

A Stability Indicating Reverse Phase High Performance Liquid Chromatography Method for Related Substances of Sofosbuvir in Tablet Dosage Form

Sofosbuvir is an antiviral drug that is used to treat hepatitis C. A present investigation deals with simple, sensitive, rapid, precise and accurate reverse phase high-performance chromatography (RP-HPLC) method developed and validated for related substances of sofosbuvir in tablet dosage form. The chromatographic separation was achieved on a Kromasil 100 C18 (250 × 4.6 mm, 5 μ) column. A sofosbuvir and its impurities were extracted by composed mixture of Mobile Phase A: buffer solution: acetonitrile (97.5:2.5% v/v) and Mobile Phase B: acetonitrile, isopropyl alcohol, methanol and purified water (60:20:10:10 % v/v/v/v) using with flow rate was 1-mL/min, column temperature was 25°C, injection volume was 10 μL, Vial thermostat temperature was 10°C. The UV detection was carried out at 263 nm. The retention time of sofosbuvir, methyl ester and ethyl ester were 54.28, 36.31 and 43.77, respectively. The method shows linearity with correlation coefficient of sofosbuvir and its impurity was 0.999 over the 0.5–7.5 ppm range. The average recovery was found to be 90.2–113.9%. The LoD and LoQ for sofosbuvir and its impurities were found to be 0.1 and 0.5 μg/mL, respectively. The method was validated as per ICH guidelines. The developed method was precise, accurate, novel and detectable towards sofosbuvir and its impurity. This method is efficient in separating the sofosbuvir and its impurity. Hence, the proposed method can be utilized for the determination of related substances in routine analysis in quality control department of pharmaceutical Industry.

Enhancing enantiomeric and diastereomeric separations: A single chromatograph approach for two-dimensional heart-cut methodology

A high-performance chromatograph was adapted and reconfigured to establish a two-dimensional methodology for separations that are not easily achievable using a single enantioselective column. Allethrin, a pyrethroid used widely as a pesticide in domestic environments, was chosen as the model analyte. With three chiral centers, it has eight stereoisomers and is considered one of the more difficult chiral separations. Diastereomeric separation was achieved in the first dimension using a shape-selective stationary phase based on ultrapure porous graphitic carbon (particle diameter, 3 μm) in the reversed phase mode. Advanced asymmetric bidirectional peak functions (half-Gaussian modified Gaussian and exponentially modified Gaussian) were employed to extract overlapping peak areas. The four diastereomeric peaks were heart-cut and sent to the second dimension online using a standard six-port 2-position rotary valve. Enantiomeric separation was attained in the second dimension using an immobilized-cellulose tris(3,5-dichlorophenylcarbamate) column (3 μm) with chromatographic resolutions >2.7 for all enantiomeric pairs. The second dimension method was developed using the information from circular dichroism detection. The baseline separation of seven peaks of allethrin in a single dimension was also demonstrated using (2-hydroxypropyl)-β-cyclodextrin. The enantiomeric purity of a purified formulation of allethrin in a commercial pesticide was validated using the developed 2D approach.

Cyclodextrin Incorporation into Covalent Organic Frameworks Enables Extensive Liquid and Gas Chromatographic Enantioseparations.

The separation of enantiomers using high-performance chromatography technologies represents great importance and interest. In this aspect, β-cyclodextrin (β-CD) and its derivatives have been extensively studied as chiral stationary phases (CSPs). Nevertheless, β-CD that was immobilized on a traditional matrix often exhibited low stabilities and limited operating ranges. Recently, covalent organic frameworks (COFs) with highly ordered nanopores are emerging as promising CSPs for enantioseparations, but their practical applications are still hampered by the difficulty of monomer and COF synthesis. Herein, two β-CD-driven COFs are synthesized via a fast and facile plasma-induced polymerization combined postsynthesis modification strategy. The precisely defined COF channels enhanced the accessibility of the accommodated β-CD to the analytes and acted as robust protective barriers to safeguard the β-CD from harsh environments. Therefore, the β-CD-modified COFs can be potentially general CSPs for extensive enantioseparation in both gas chromatography and high-performance liquid chromatography, and a wide range of racemates were separated. Compared to the commonly employed commercial chiral columns, these COF-based columns exhibited comparable resolution capability and superior application versatility. This work integrates the advantages and overcomes the defects of COFs and β-CD, thus advancing COFs as platforms for chiral selector modification and giving great promise for practical chromatographic enantioseparation.

Potential Antioxidant and Anti-Inflammatory Effects of Lonicera japonica and Citri Reticulatae Pericarpium Polyphenolic Extract (LCPE).

Dermatitis is an inflammatory condition of the outer layer of the skin that causes itching, blisters, redness, swelling, and often exudation, scabs, and peeling. Among them, purulent inflammation is a symptom that often occurs on the skin and appears in the form of boils and acne. Various studies are being conducted to treat these inflammatory diseases. Accordingly, Lonicera japonica and Citri Reticulatae Pericarpium Polyphenolic Extract (LCPE), which uses herbal preparations such as Lonicera japonica, Citri Reticulatae Pericarpium, and Glycyrrhiza uralensis, has been used to suppress inflammation since ancient times, and its anti-inflammatory effect can be observed in skin keratinocytes after inducing inflammation. In this study, the major polyphenolic compounds in LCPE were quantitatively determined by analyzing the data through peak values using high-performance chromatography (HPLC-MS/MS) coupled with mass spectrometry. Additionally, bioactive compounds targeting 2,2-diphenyl-1-picrylhydrazyl (DPPH) were analyzed by ultrafiltration integrated with LC. Several compounds with the most significant effects were selected (chlorogenic acid, narirutin, and isorhamnetin). Skin keratinocytes induced by lipopolysaccharide (LPS) were treated with LCPE to show its anti-inflammatory effects. After LCPE treatment, inflammation-mediating cytokines such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) were decreased. In addition, nuclear factor kappa (NF-кB) and mitogen-activated protein kinase (MAPK) were inhibited in important pathways related to inflammation. Lastly, molecular modeling was performed to determine binding scores with inflammation-related proteins using molecular docking for the selected compounds. According to these results, LCPE is effective in treating keratinocytes induced by LPS and reducing inflammation and has potential antioxidant effects, and the polyphenol components have been identified.

Oil palm kernel globulin antihypertensive peptides: isolation and characterization, ACE inhibition mechanisms, zinc-chelating activity, security and stability.

Introduction: The oil palm kernel (OPK) expeller is the main byproduct of palm oil, but its utilization is limited. Methods: To obtain angiotensin-I-converting enzyme (ACE) inhibition peptides with Zn-chelating capacity, defatted oil palm kernel globulin hydrolysates (DOPKGH) were subjected to Sephadex G-15 gel electrophoresis, reverse-phase high liquid performance chromatography, and UPLC-ESI-MS/MS analysis. Results and discussion: Five representative oligopeptides, including Gln-Arg-Leu-Asp-Arg-Cys-Lys (QRLERCK), Leu-Leu-Leu-Gly-Val-Ala-Asn-Tyr-Arg (LLLGVANYR), Arg-Ala-Asp-Val-Phe-Asn-Pro-Arg (RADVFNPR), Arg-Val-Ile-Lys-Tyr-Asn-Gly-Gly-Gly-Ser-Gly (RVIKYNGGGSG), and Glu-Val-Pro-Gln-Ala-Tyr-Ile-Pro (EVPQAYIP), without potential toxicity and allergenicity, were identified in DOPKGH. Of these, only EVPQAYIP showed both ACE-inhibitory activity (IC50: 102.75μmol/L) and Zn-chelating capacity (11.69mg/g). Molecular docking and inhibition kinetics showed that EVPQAYIP was a competitive inhibitor of ACE because it could bind to Glu384, Lys511, and Gln281 (belonging to the central S1 and S2 pockets, respectively) of ACE. Moreover, EVPQAYIP affects zinc tetrahedral coordination in ACE by binding to Glu411; the amino and carboxyl groups of EVPQAYIP chelate with zinc ions. During gastrointestinal digestion, the ACE inhibitory activity of EVPQAYIP was relatively stable. Additionally, EVPQAYIP enhanced zinc stability in the intestine and exerted antihypertensive effects in spontaneous hypertensive rats. These results suggest the potential application of OPK peptides as ingredients in antihypertensive agents or zinc fortification.

Selenocysteine Formation by Enterococcus faecium ABMC-05 Follows a Mechanism That Is Not Dependent on Genes selA and selD but on Gene cysK

Lactic acid bacteria (LAB) resist sodium selenite of concentrations greater than 100 mg/L in fermentation media. Selenium affects the growth rate, but once the microorganism absorbs selenium, this element is converted through a complex mechanism into selenocysteine and then into a selenoprotein structure. This study verified the presence of selenocysteine in Enterococcus faecium ABMC-05. The microorganism was cultivated in a medium enriched with a minimum inhibitory concentration of sodium selenite (184 mg/L). The concentration of selenium absorbed and the bioconversion into selenocysteine were determined by inductively coupled plasma optical emission spectrometry (ICP-OES) and reverse-phase high-performance chromatography (RP-HPLC), respectively. The presence of the selD, selA, and cysK genes was determined by amplifying the 16S rDNA through polymerase chain reaction (PCR). The microorganism accumulated inorganic selenium, and part was transformed into selenocysteine. The growth curves were atypical for a lactic acid bacterium with a stationary phase greater than 70 h. Determining the genetic expression showed only the presence of the cysK gene and the absence of the selD and the selA genes. The results demonstrate that this microorganism produces selenocysteine through a mechanism independent of the SelA and SelD pathways in contrast to other LAB.

Detection of some Volatile Oils using High-Performance Chromatography Technique and Study Antibacterial Activity of Black Seed Extract Nigella sativa L.

The antibacterial activity of crude extract of Nigella sative extract was studied on the some pathogenic bacteria by using paper disc diffusion. The tested strains were: Escherichia coli, Enterococcus faecalis, staphylococcus aureus (MRSA) and psendomonas aeruginosa. The results were showed that the crude extract produced inhibition zone in all tested bacteria ranging from (20 mm) in S.aureus and MRSA and (12 mm) in P.aureginusa. Some Volatile Oils were determined using a high-performance liquid chromatograph(hplc), and three extraction methods (aqueous, methanol and ethanol) were used. The following Volatile Oils were estimated ((Limonene, Anethole, -α-pinene, Estagole) reached(35.61,86.40,41.69,35.61) sequentially While the aqueous extraction recorded the lowest averages of (30.52, 71.42, 32.82, 25.89). In conclusion, the current article was attitude to assess the effectiveness of N. sativa in front of all prevalent illnesses caused by strains of microorganisms that are drug-resistant.

Development, Validation and Approbation Analytical Method for the Quantitative Determination of Taurine by HPLC-UV Method in the Test of Comparative Dissolution Kinetics

Introduction. Taurine is a non-proteinogenic amino acid. The molecule is involved in lipid metabolism, adsorbs fat-soluble vitamins, and its conjugates with bile acids contribute to the emulsification of fats in the intestine. Drugs, which include a taurine molecule, have anti-cataract, cardiotonic, metabolic effects, and also stimulate regeneration. Among the dosage forms, where taurine acts as an active substance, there is a solid dosage form – film-coated tablets. One of the methods for assessing the quality of solid dosage forms is a comparative dissolution kinetics test. High-performance chromatography with ultraviolet detection is a widely used method for quantification within the dissolution test, however, for taurine, which does not contain chromophore groups in its structure, this method is not directly applicable. To solve this problem, one can apply the method of pre-column derivatization, because of which an fragment is introduced into the structure, providing a bathochromic shift in the UV spectrum of the starting compound.Aim. Development, validation and approbation analytical method for the quantitative determination of taurine by high-performance chromatography with ultraviolet detection as part of a test comparative kinetics dissolution of taurine tablets with a dosage of 250 and 500 mg.Materials and methods. The following preparations were used for the analysis: taurine tablets, film-coated 250 mg and 500 mg, domestic production with a valid expiration date. The comparative dissolution kinetics test was carried out on a DT 126 Light instrument for the "Dissolution" test (ERWEKA GmbH, Germany). Chromatographic separation and detection were performed on a Nexera-i LC-2040 high-performance liquid chromatograph (Shimadzu Corporation, Japan) equipped with a column and sample thermostat, a degasser, an autosampler, and an ultraviolet detector. Detection was carried out at a wavelength of 254 nm after derivatization of the taurine molecule with 4-toluenesulfonyl chloride. Were used a Shim-pack Velox C18 5 μm 4.6 × 150 mm column (Shimadzu Corporation, Japan) and a Shim-pack Velox C18 EXP Guard Column Cartridge 5 μm 4.6 × 5 mm (Shimadzu Corporation, Japan). Primary data were processed using LabSolutions Single LC software (Shimadzu Corporation, Japan).Results and discussion. The optimal conditions for taurine derivatization have been selected, and a method for the quantitative determination of taurine by HPLC-UV in test comparative kinetics dissolution in three dissolution media: 0.1M hydrochloric acid solution with pH 1.2, acetate buffer solution with pH 4.5, phosphate buffer solution with pH 6.8, as well as in the quality control medium – purified water has been developed and validated. During the validation of the developed methodology, it was found that the validation characteristics are within the acceptance criteria in 4 dissolution media. The analytical range of the method was 0.05–1.2 mg/mL and allows the developed method to be used for the quantitative determination of tablets with a dosage of 250 mg and 500 mg as part of the test comparative kinetics dissolution. The method was tested in 4 dissolution media, in all media, there was a complete release in both dosages (more than 85 % by 30 minutes).Conclusion. The method was tested in three dissolution media: 0.1 M hydrochloric acid solution with pH 1.2, acetate buffer solution with pH 4.5, phosphate buffer solution with pH 6.8, as well as in the quality control medium – purified water. In all media, there was a complete release in both dosages (more than 85 % by 30 minutes).

Toxicity of the Hexane Fraction of Fruits and Seeds of Ricinus communis to Caterpillars of the Spodoptera Complex

The objective of this work was to evaluate the effect of the hexane fraction of the crude extract of the fruits and seeds of Ricinus communis (HFFSRC) on eggs of Spodoptera cosmioides and Spodoptera frugiperda and on caterpillars of Spodoptera eridania, S. frugiperda, and S. cosmioides, under laboratory conditions through topical application and ingestion, as well as to identify the compounds in the hexane fraction through high-performance chromatography (HPLC-ESI-Q-TOF-MS/MS). To do so, three bioassays were conducted: (1) the effect of HFFSRC at 2% (20,000 mg·mL−1) on eggs of S. cosmioides and S. frugiperda, (2) the lethal effect of HFFSRC at 2% applied topically, and (3) the lethal effect of HFFSRC at 2% applied to soybean-leaf discs on first-, second-, third-, and fourth-instar caterpillars of the three insect species. It was found that 2% HFFSRC had an ovicidal effect on S. frugiperda and S. cosmioides, completely reducing larvae hatching and the insecticidal effect for the four instars of the three insect species when applied topically and on food. Five compounds were identified in the HFFSRC: three flavonoids, one ricinoleic acid, and one cinnamic acid. HFFSRC at 2% had an acute ovicidal and insecticidal effect on caterpillars of the studied species, configuring itself as a potential insecticide.

Aflatoxins in raw Brazil nut (Bertholletia excelsa H.B.K.)

The Brazil nut is a nutritionally rich food, produced and consumed in tropical regions. On the other hand, it may be suitable to toxigenic fungi and consequently contamination by aflatoxins (AFL), toxic substances to consumers. In this context, the objective of this work was to evaluate the occurrence of AFL in raw Brazil nuts, that is, before being subjected to industrial drying stages. Of 23 samples were purchased at retail in the city of Manaus-Am-Brazil and evaluated by high performance and liquid chromatography for total AFL (B1+b2+G1+G2). From the samples, 8.6 % were positive for total AFL and. Only AFG1 and AFG2, were detected in the LOD (0.8 μg/kg) and met to the limit of &lt;10 µg/kg of current legislation in Brazil and European Union. In conclusion, despite the contaminated samples met the legal limits, that seems the AFL contamination could happen in raw Brazil nut, as it has been reported in dried seeds. In this sense, some effort in the initial steps of the Brazil nut chain must be applied to mitigate AFL contamination before reach the factory processing.

Biologically active compounds in Scutellaria baicalensis L. callus extract: Phytochemical analysis and isolation

Plant cells and tissue cultures are sources of secondary plant metabolites. Substances produced by callus cultures can expand the raw material base in pharmacy and food production. However, isolating biologically active substances from medicinal plants is a labor- and time-consuming process. As a result, new and efficient technological processes adapted for extraction from callus cultures are in high demand, and new algorithms of isolation and purification of biologically active substances remain a relevant task.&#x0D; This research featured callus cultures of Scutellaria baicalensis. The procedures for phytochemical analysis and isolation of biologically active substances involved such physicochemical research methods as high-performance chromatography (HPLC), thin-layer chromatography (TLC), UV spectrometry, and IR spectrometry.&#x0D; The high performance liquid chromatography confirmed the presence of flavonoids represented by baicalein (5,6,7-trioxyflavone), baicalin (baicalein 7-O-glucuronide), scutellarein (5,6,7,4-tetraoxyflavone), scutellarin (7-O-glucuronide scutellarein), vagonin, and oroxylin. The spectral analyses also detected skutebaicalin. The highest total content of diterpene belonged to the samples extracted with 70% ethanol at 70°C. The content of diterpene was 0.09 mg/cm3 in terms of betulin. The biologically active substances were isolated from the callus extracts of S. baicalensis with a recovery rate of ≥ 80%. The purification scheme made it possible to obtain highly-pure individual biologically active compounds: trans-cinnamic acid, baicalin, and oroxylin A had a purity of ≥ 95%; baicalein had a purity of ≥ 97%; scutellarin and luteolin reached ≥ 96%.&#x0D; The new technological extraction method made it possible to obtain extracts from S. baicalensis callus cultures, which were tested for the component composition. The developed isolation algorithm and purification scheme yielded biologically active substances with a purification degree of ≥ 95%.

 Dissipation kinetics and dietary risk assessment of chlorantraniliprole residues in/on garden bean (Lablab purpureus var. typicus) using ultra high-performance liquid chromatography 

The aim of the study was to find out the dissipation rate of the chlorantraniliprole residues in garden bean pods. Insecticides were sprayed at the farmers’ holding located at Naraseepuram, Coimbatore during the year 2022 (March to April), and the collected samples were processed and analyzed in the laboratory. An effective modified QuEChERS method was developed and validated to estimate the chlorantraniliprole residues in Ultra High-Performance Chromatography. Good linearity with R2 greater than 0.99 was achieved with the Limit of Quantification as 0.05 mg/kg. The relative standard deviation was below 5% for the recoveries obtained (93.30 to 97.91%). The residues persisted up to 10 and 15 days after treatment for recommended (20 g a.i./ha) and double the recommended doses (40 g a.i./ha), respectively. The half-life of chlorantraniliprole in garden beans was calculated as 2.18 days for recommended dose and 2.89 days for double the recommended dose. Intake of chlorantraniliprole residues through garden beans at either of the doses was found to be safe as the calculated Risk Quotient (RQ) was less than one.