Thin-layer Chromatography Research Articles

Synthesis, Characterization, and Optimization of Novel Furan-ring Fused Chalcones via Radical Cyclization of α,β-Unsaturated Ketones and Cyclic Ketone

Abstract: Novel Furan-ring Fused Chalcones (FFC) were synthesized using a radical cyclization reaction of α,β-unsaturated ketones with cyclic ketone as the model reaction to attain this goal. In this study, traditional and microwave-assisted methods for the efficient and cost-effective synthesis of furan-ring fused chalcones in mild reaction conditions are compared and optimized. The goal is to develop a reliable and adaptable synthetic technique that may be used to produce these useful chalcone derivatives quickly and effectively. The optimal experimental conditions for these reactions were carefully determined using two independent methodologies: conventional (Method A) and microwave (Method B). The results indicated that the proposed method B could be used effectively in the future to synthesize novel furans with short reaction times and acceptable yields (87-94 %), and products were purified by column chromatography and preparative thin layer chromatography (PTLC). All new compounds were characterized by 1H-NMR, 13C-NMR, LC-MS, and elemental analyses.

Innovative antibacterial and antifungal coating films of chitosan/nano zinc oxide/essential oil on grape preservation

The effects of some innovative antibacterial and antifungal coating films of chitosan/nano zinc oxide/essential oil (C-ZNO-EO) on grape preservation for human health were studied during the storage periods on (0, 4, 7, 14, and 16) days. Several evaluation approaches were evaluated such as the bacterial evaluations against the presence of the mesophilic aerobic bacteria. The polyamines evaluation was studied by thin-layer chromatography (TLC) for putrescine and spermidine. Soluble sugars were detected by high-performance liquid chromatography (HPLC) for fructose, glucose, and sucrose. Iron, zinc, and manganese were absorbed by the spectrum spectrophotometer. The antifungal properties have been examined against Botryis cineraa, Aspergillus niger, and Tubingensis. The results reported that the highest mesophilic aerobic bacteria (4.73 Log CFU g−1) was recorded in the control grapes on the 16th day, whereas the lowest level (2.05 Log CFU g−1) was recorded for C-ZNO-EO. The polyamines ranged from 12.18 nmol g−1 FW to 10.1 nmol g−1 FW, and from 21.35 nmol g−1 FW to 16 nmol g−1 FW for putrescine and spermidine during the storage periods, respectively. The grapes treated with C-ZNO-EO achieved the maximum increase in glucose and fructose contents (40.61 and 30.05 μmol/g FW), while grapes treated with chitosan/nano zinc oxide (C-ZNO) achieved the maximum increase in sucrose (0.982 μmol/g FW) compared with chitosan (C) and control treatments at the end of the storage periods, respectively. Using C-NZO-EO treatment had a positive effect on the mineral compositions and the antifungal against Botryis cineraa, Aspergillus niger, and Tubingensis. The innovative antibacterial and antifungal coating film by C-NZO-EO was the best treatment for grape preservation.

A novel stability indicating HPTLC method for comparative quantitative estimation of quercetin in herbal preparations

This research introduces a novel stability-indicating HPTLC (High-Performance Thin-Layer Chromatography) method for the comparative quantitative estimation of Quercetin in herbal preparations. Through extensive stability studies, the method is validated, providing a robust framework for evaluating Quercetin's stability in diverse herbal formulations. The developed method ensures accurate and reliable quantification of Quercetin, facilitating comparative analysis across various herbal preparations. Additionally, results reveal that Quercetin degraded under most tested conditions, such as acidic, alkaline, oxidative and neutral conditions underscoring the necessity for intervention by experts to eliminate such challenges and ensure product efficacy and quality in the herbal industry.

Comparative study of Scleromitrion diffusum and Oldenlandia corymbosa: Microscopy, TLC, HPLC, and antioxidant activity.

Quality control of herbal medicines is crucial, especially the role of herbal drug identification. This is essential for preventing the misuse of herbs, which can affect efficacy or cause toxicity. Scleromitrion diffusum is a common herb, yet it is often mistaken for Oldenlandia corymbosa. This study analyzed the morphology, microscopy, thin-layer chromatography (TLC), and high-pressure liquid chromatography (HPLC) using two markers, asperuloside and scandoside methyl ester, to distinguish between S. diffusum and O. corymbosa with the analysis included 10 samples of S. diffusum and 10 samples of O. corymbosa collected from the Taiwan market. By quantifying the total polyphenols and flavonoids, we investigated the antioxidant activity, including the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging effect, 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+) scavenging effect, and reducing power to further elucidate the biological effects of the two herbs. The results of this study revealed notable differences in microscopy and suggested a TLC method for distinguishing between the two herbs in the market. In HPLC, the ratios of asperuloside and scandoside methyl ester differed between the two herbs. S. diffusum contained a higher asperuloside content. In contrast, O. corymbosa contained higher concentrations of scandoside methyl esters. With more total polyphenols and flavonoids in S. diffusum than those in O. corymbosa, the antioxidant activity of S. diffusum was superior to that of O. corymbosa. This study provides a comprehensive understanding for the identification and quality evaluation of S. diffusum in the market. RESEARCH HIGHLIGHTS: The study consolidates and clarifies the morphological and microscopic differences between Scleromitrion diffusum and Oldenlandia corymbosa - a common adulterant species of S. diffusum on the Taiwan markets. Using Asperuloside and Scandoside methyl ester as two chemical markers, the study proposes a TLC method for rapidly testing S. diffusum and O. corymbosa on the market. Through HPLC analysis, our results showed that S. diffusum and O. corymbosa had a clear difference in the ratio of two markers, Asperuloside and Scandoside methyl ester: Asperuloside/Scandoside methyl ester in S. diffusum is higher than that in O. corymbosa. Through phytochemicals contents, including total phenols content, flavonoids content, and antioxidant activity, including DPPH, ABTS•+ scavenging activity, and reducing power, S. diffusum showed slightly higher levels of phenols and flavonoids as well as a better antioxidant activity than O. corymbosa.

Isolation and identification of hyperin and naringenin from guava cultivated in Iraq and evaluation of cytotoxic activity of hyperin

The guava plant, Psidium guajava L., serves as proof of the abundant donations of nature, providing a delicious guava fruit; this plant is rich in groups of medicinal and nutritional benefits. Guava belonging to the Myrtaceae family, many previous studies reported many phytochemical constituents in its leaves that have many pharmacological activities and medicinal properties; this study focuses on the isolation, structural elucidation and calculation concentration of flavonoids, assessment of the cytotoxic activityof hyperin from Psidium guajava leaves newly cultivated in Iraq. The isolation process involved the use of thin-layer chromatography (TLC) and preparative high-performance liquid chromatography (PHPLC) and structural elucidation involved NMR (nuclear magnetic resonance spectroscopy) and FTIR (Fourier transform infrared spectrometer) for detailed structural insights into 2 isolated flavonoids. The isolation techniques proved effective in obtaining pure samples of hyperin and naringenin from the ethyl acetate and n.butanol fractions and structural elucidation techniques gives a good explanation for 2 isolated flavonoids. Evaluate cytotoxic activity of hyperin flavonoid against prostate cancer cell line (PC-3), human breast cancer cell line (MCF-7) and normal dermal fibroblast neonatal (HdFn), hyperin flavonoid exhibited a decrease in cell viability (%) with IC50 58.9 ?g /mL against the prostate cancer cell and IC50 90.58 ?g /mL against human breast cancer and noncytotoxic to HDFn, with an IC50 value substantially surpassing concentrations of 100 ?g/mL. These analytical approaches provided a comprehensive understanding of the chemical composition of the isolated flavonoids and interpreted that guava has cytotoxic activity against some cancers, depending on a concentration-dependent mode.

Anti-Inflammatory and Anti-Arthritis Activity of Quercetin: A Comprehensive Review.

This comprehensive exploration delves into the multifaceted attributes of quercetin, a flavonoid with extensive health-promoting potential. The review navigates through its fundamental properties, encompassing its chemical structure, classification as a flavonoid, and its natural prevalence in various sources. Addressing solubility, stability, and bioavailability challenges, the investigation delves into innovative isolation techniques, including solvent extraction, solid-phase extraction, natural deep eutectic solvents, supercritical fluid extraction, microwave-assisted extraction, column chromatography, and high-performance thin-layer chromatography. Transitioning into pharmacological implications, the study unveils quercetin's roles in anti-inflammatory pathways, antioxidant effects, and immune modulation, reflecting its versatile significance in health management. The review highlights its impact on wound healing processes and its potential to mitigate arthritis, elucidating its holistic contributions. Culminating in an exploration of recent studies, the analysis underscores quercetin's remarkable anti-inflammatory and anti-arthritis activities, reflecting its substantial potential across various ailments. The review concludes by projecting future trajectories, emphasizing prospects for an advanced understanding of quercetin's mechanisms, sustainable extraction techniques, clinical integration, and exploration of synergistic combinations. Collectively, this review investigation underscores quercetin's dynamic role at the intersection of natural compounds and medicinal applications, offering profound implications for well- being and health enhancement.

Phytochemical Analysis, Antioxidant Potential and Antibacterial Activities of Different Anatomical Parts of Hypericum cordifolium Choisy

The genus Hypericum comprises a large number of species. The flower, leaf, stem, and root of the Hypericum species are widely used in traditional medicine in different cultures. Many Hypericum species have been well investigated phytochemically and pharmacologically. However, only a few reports are available on the H. cordifolium native to Nepal. The present study aims to evaluate the phytochemical composition of different extracts, qualitative analysis of methanol extract of the flower and leaf using thin-layer chromatography (TLC), and the antioxidant properties of components by the TLC-DPPH. assay. The phenolic and flavonoid contents were estimated in different extracts of the leaf and stem, and their antioxidant and antibacterial activities were evaluated. In the phytochemical screening, phenolics and flavonoids were present in ethyl acetate, methanol, and 50% aq methanol extracts of both the leaf and stem. In TLC analysis, the methanol extract of flowers showed the presence of 11 compounds and the leaf extract showed the presence of 8 compounds. Both extracts contained chlorogenic acid and mangiferin. Hyperoside and quercetin were present only in the flower extract. In the TLC-DPPH. assay, almost all of the flower extracts and 5 compounds of the leaf extract showed radical scavenging potential. Estimation of phenolics and flavonoids showed that all the leaf extracts showed higher amounts of phenolics and flavonoids than stem extracts. Among leaf extracts, greater amounts of phenolics were detected in 50% aqueous methanol extract (261.25 ± 1.66 GAE/g extract) and greater amounts of flavonoids were detected in methanol extract (232.60 ± 10.52 CE/g extract). Among stem extracts, greater amounts of flavonoids were detected in the methanol extract (155.12 ± 4.30 CE/g extract). In the DPPH radical scavenging assay, the methanol extract of the leaf showed IC50 60.85 ± 2.67 µg/ml and 50% aq. methanol extract of the leaf showed IC50 63.09 ± 2.98 µg/ml. The methanol extract of the stem showed IC50 89.39 ± 3.23 µg/ml, whereas ethyl acetate and 50% aq. methanol extract showed IC50 > 100 µg/ml. In the antibacterial assay, the methanol extract of the leaf showed the inhibition zone of 12-13 mm and the stem extract showed the inhibition zone of 7–11 mm against S. aureus, E. coli, and S. sonnei, whereas both extracts were inactive against S. typhi. The findings of this study support the traditional use of this plant in Nepal for the treatment of diseases associated with bacterial infections. The present study revealed that the underutilized anatomical parts of H. cordifolium could be the source of various bioactive phytochemicals like other Hypericum species.

Comparison of HPLC, HPTLC, and In Silico Lipophilicity Parameters Determined for 5-Heterocyclic 2-(2,4-Dihydroxyphenyl)-1,3,4-thiadiazoles

The 5-heterocyclic 2-(2,4-dihydroxyphenyl)-1,3,4-thiadiazoles were obtained as potential biologically active compounds. Lipophilicity is one of the most important physicochemical properties of compounds and was already taken into account during the drug candidates design and development. The lipophilicity of compounds was determined using the computational (log P) and chromatography (log kw, RMw) methods. The experimental ones included the reverse-phase column high performance liquid chromatography RP (HPLC) with C8, C18, phosphatidylcholine (IAM), and cholesterol stationary phases and the thin layer chromatography (RP-HPTLC) with C8 and C18 stationary phases and various organic modifiers under the isocratic conditions. Descriptive statistics, correlation, and PCA analyses were used to compare the obtained results. For lipophilicity estimation of the tested compounds by HPTLC, dioxane and MeOH seem to be particularly beneficial as organic modifiers. The chromatographic lipophilicity parameters log kw (RMw) were well correlated and highly redundant (85%) compared with those calculated. Most compounds possess lipophilicity parameters within the recommended range for drug candidates.

FERMENTATION, PURIFICATION AND IN-VITRO EVALUATION OF ANTIMICROBIAL, ANTITUMOR AND ANTIOXIDANT COMPOUNDS FROM A HIMALAYAN ACTINOBACTERIA STRAIN STREPTOMYCES SP. PU-AK14

This study reports the laboratory scale fermentation, compounds purification and bioactivity screening of the strain Streptomyces sp. PU-AK14 isolated from Himalayan mountains range, Pakistan, for its potential to produce antimicrobial, antioxidant and antitumor compounds. For this purpose, laboratory fermentation (20 L working volume) was performed followed by extraction of the compounds by XAD adsorbent gel using the solvents methanol and ethyl acetate. Further purification of the compounds was done by silica gel column chromatography, sephadex column and preparative TLC. A total of 18 partially purified compounds were retrieved from crude extract and were screened for biological activity. The crude extract was more active against gram negative bacteria with MIC of 0.1 mg/ml against Klebsiella pneumoniae and Escherichia coli. Eight of the partially purified compounds exhibited activity against MRSA whereas five fractions were active against gram negative bacteria. Seven partially purified fractions exhibited promising antioxidant activity with radical scavenging activity ranging from 78.2 to 42.34 percent. Cytotoxicity of partially purified compounds ranged from 75% to 100% larval mortality at 100mg/ml against Artemia salina and 50% to 52% cell mortality against HCT 116 colorectal cancer cell line at 100mg/ml. The best anti-tumor activity was exhibited by the fraction AK141111L. The study suggested that Streptomyces sp. PU-AK14 is a potential source of chemotherapeutically useful compounds.

Phytochemical and Antioxidant Investigations of Extracts from the Leaves of Macaranga heterophylla (Müll. Arg.) (Euphorbiaceae), A Medicinal Species used in Côte d'Ivoire

Aims: This work aims to investigate the phytochemical composition and antioxidant potential of the leaves of Macaranga heterophylla. Methodology: For this purpose, phytochemical screening by detection tests and thin layer chromatography (TLC), determination of total phenols content, total flavonoids content and condensed tannins content, and assessment of antioxidant potential by DPPH and reducing power tests were carried out on aqueous crude extracts, ethanolic crude extracts and selective extracts of the leaves of M. heterophylla. Results: The percentage yields obtained with ethanol (70%) (28.90 and 24.70% for the ethanolic decoction and ethanolic macerate respectively) are higher than those obtained with water (24.30 and 21.10% for the aqueous decoction and aqueous macerate respectively). The phytochemical screening highlighted the presence of several phytochemical families such as phenolic compounds (coumarins, flavones, tannins), quinones, sterols and polyterpenes, saponosides, glycosides, cardiotonic glycosides and oligosaccharides. Quantitative analysis of total phenolics, total flavonoids and proanthocyanidols showed that their respective levels in the leaves of M. heterophylla varied depending on the solvent and the extraction technique. Concerning total phenolics, the aqueous decoction and ethanolic macerate gave the best total phenol contents (129.04 ± 9.53 and 119.82 ± 2.63 mg EAG/g DM respectively); for total flavonoids, the aqueous decoction gave the best content (33.03 ± 1.61 mg EQ/g DM), while for condensed tannins, the aqueous macerate gave the best content (0.87 ± 0.02 mg ECT/g DM). With regard to DPPH antioxidant activity, the results showed that the ethanolic decoction has more pronounced antioxidant activity than the aqueous decoction, while the aqueous macerate showed better antioxidant activity than the ethanolic macerate. Concerning the reducing power test, the opposite trend was observed. Conclusion: The present study demonstrated that M. heterophylla is a concentrate of secondary metabolites with antioxidant properties, which would explain its use in traditional medicinal practice.

Characterization of antibiofilm compound from marine sponge Stylissa carteri.

The fouling phenomenon grabbed global attention and caused huge economic losses specifically in marine-related industries. Sessile behavior exposed the sponge to the risk of fouling. However, their bodies remained free from foulers, which were attributed to the chemical defense system. The objectives of this study were to determine the antibiofilm activity of the marine sponge, Stylissa carteri, and to characterize the isolated compound involved. The antibiofilm activity of S. carteri methanolic crude extract (MCE) and fractions was tested against biofilm-producing bacteria, Pseudomonas aeruginosa, using two different modes of crystal violet biofilm assays: preventive and detachment. Besides that, the disc-diffusion test was conducted to screen the antibacterial activity against gram-positive and gram-negative bacteria while a cytotoxicity assay was conducted on the HepG2 cell line. Bioassay-guided fractionation was carried out using vacuum liquid chromatography (VLC) and solid phase extraction using a C18 Sep-Pak Cartridge. The crystal compound was isolated and characterized through thin-layer chromatography (TLC), Fourier transform infrared (FTIR) spectroscopy, liquid chromatography-mass spectrometry (LCMS), and nuclear magnetic resonance (NMR) spectroscopy. The S. carteri MCE showed a promising result with a half-maximal inhibitory concentration (IC50) of 20.22μg/mL in the preventive assay, while no IC50 was determined in the detachment assay since all inhibitions < 50%. The S. carteri MCE exhibited broad-spectrum antibacterial activity and displayed a non-cytotoxic effect. Fraction 4 from MCE of S. carteri (IC50 = 2.40μg/mL) reduced the biofilm in the preventive assay at all concentrations and exhibited no antibacterial activity indicating the independence of antibiofilm from antibacterial properties. Based on the data obtained, an alkaloid named debromohymenialdisine (DBH) was identified from Fraction 4 of S. carteri MCE. In conclusion, S. carteri was able to reduce the establishment of the biofilm formed by P. aeruginosa and could serve as a prominent source of natural antifouling agents.

Development of White Cabbage, Coffee, and Red Onion Extracts as Natural Phosphodiesterase-4B (PDE4B) Inhibitors for Cognitive Dysfunction: In Vitro and In Silico Studies.

Human cognition fundamentally depends on memory. Alzheimer's disease exhibits a strong correlation with a decline in this factor. Phosphodiesterase-4 B (PDE4B) plays a crucial role in neurodegenerative disorders, and its inhibition is one of the promising approaches for memory enhancement. This study aimed to identify secondary metabolites in white cabbage, coffee, and red onion extracts and identify their molecular interaction with PDE4B by in silico and in vitro experiments. Crushed white cabbage and red onion were macerated separately with ethanol to yield respective extracts, and ground coffee was boiled with water to produce aqueous extract. Thin layer chromatography (TLC)-densitometry was used to examine the phytochemicals present in white cabbage, coffee, and red onion extracts. Molecular docking studies were performed to know the interaction of test compounds with PDE4B. TLC-densitometry analysis showed that chlorogenic acid and quercetin were detected as major compounds in coffee and red onion extracts, respectively. In silico studies revealed that alpha-tocopherol (binding free energy (∆Gbind) = -38.00 kcal/mol) has the strongest interaction with PDE4B whereas chlorogenic acid (∆Gbind = -21.50 kcal/mol) and quercetin (∆Gbind = -17.25 kcal/mol) exhibited moderate interaction. In vitro assay showed that the combination extracts (cabbage, coffee, and red onion) had a stronger activity (half-maximal inhibitory concentration (IC50) = 0.12 ± 0.03 µM) than combination standards (sinigrin, chlorogenic acid, and quercetin) (IC50 = 0.17 ± 0.03 µM) and rolipram (IC50 = 0.15 ± 0.008 µM). Thus, the combination extracts are a promising cognitive enhancer by blocking PDE4B activity.

Linking Chemical Structure to the Linear and Nonlinear Properties of Asphalt Binders

The study described in this paper aims to establish the relationship between the chemical composition and rheological properties of asphalt binders in both the linear viscoelastic (LVE) and nonlinear viscoelastic (NLVE) domains. Two asphalt binders with different penetration grades were subjected to four distinct aging treatments of varying severity, resulting in changes in their chemical fingerprints. Chemical characteristics of the asphalt binders were analyzed using thin-layer chromatography (TLC), Fourier Transform infrared spectroscopy (FTIR), and gel permeation chromatography (GPC). Frequency sweep tests were conducted to evaluate the LVE behavior of the asphalt binders, while multiple stress creep recovery (MSCR) tests were employed to assess their NLVE properties. With regard to the LVE properties, rheological index ( R) and zero shear viscosity (ZSV) derived from the Christensen-Anderson-Marasteanu (CAM) model exhibited high correlations with FTIR and molecular weight distribution (MWD) parameters. Additionally, the polydispersity index (PDI) displayed a stronger correlation with LVE-based parameters. Findings from the MSCR tests revealed that the sensitivity of the percentage recovery ( %R) to chemical composition, as opposed to nonrecoverable creep compliance ( Jnr), can be largely attributed to the degree of nonlinearity. Furthermore, it was observed that lower molecular weight molecules exert a greater influence on %R in the nonlinear domain. Finally, it was found that Jnrslope is a more reliable parameter than Jnrdiff for assessing the effects of chemical makeup on stress and temperature sensitivity.

Neurospora sp. Mediated Synthesis of Naringenin for the Production of Bioactive Nanomaterials.

The application of Neurospora sp., a fungus that commonly thrives on complex agricultural and plant wastes, has proven successful in utilizing citrus peel waste as a source of naringin. A UV-Vis spectrophotometric method proved the biotransformation of naringin, with an absorption maximum (λmax) observed at 310 nm for the biotransformed product, naringenin (NAR). Further verification of the conversion of naringin was provided through thin layer chromatography (TLC). The Neurospora crassa mediated biotransformation of naringin to NAR was utilized for the rapid (within 5 min) synthesis of silver (Ag) and gold (Au) nanoconjugates using sunlight to accelerate the reaction. The synthesized NAR-nano Ag and NAR-nano Au conjugates exhibited monodispersed spherical and spherical as well as polygonal shaped particles, respectively. Both of the nanoconjugates showed average particle sizes of less than 90 nm from TEM analysis. The NAR-Ag and NAR-Au nanoconjugates displayed potential enhancement of the antimicrobial activities, including antibacterial and nematicidal properties over either standalone NAR or Ag or Au NPs. This study reveals the potential of naringinase-producing Neurospora sp. for transforming naringin into NAR. Additionally, the resulting NAR-Ag and NAR-Au nanoconjugates showed promise as sustainable antibiotics and biochemical nematicides.

Simultaneous determination of amlodipine besylate and azilsartan mixture in human plasma utilizing high-performance thin-layer chromatography with ultraviolet detection

Simultaneous determination of amlodipine besylate and azilsartan mixture in human plasma utilizing high-performance thin-layer chromatography with ultraviolet detection

Fagopyrin F fraction from Fagopyrum tataricum demonstrates photodynamic inactivation of skin infecting bacterium and squamous cell carcinoma (A431) cells.

Photodynamic therapy (PDT) stands out as a noteworthy development as an alternative targeted treatment against skin ailments. While PDT has advanced significantly, research into photo-activatable "Green drugs" derived from plants which are less toxic than the synthetic drugs has not kept pace. This study investigates the potential of Fagopyrin F Containing Fraction (FCF) derived from Fagopyrum tataricum in mediating PDT against Staphylococcus aureus and skin cancer cells (A431). FCF was isolated from the plant extract using thin-layer chromatography, followed by identification of the compound through high-performance liquid chromatography and high-resolution liquid chromatography-mass spectrometry. FCF was tested to determine its antibacterial and anticancer efficacy. Results revealed that FCF-mediated PDT exhibited potent action against S. aureus, significantly reducing bacterial viability (MIC 19.5 μg/100 μL). Moreover, FCF-mediated PDT showed good efficacy against A431 cells, resulting in a notable reduction in cell viability (IC50 29.08μg/mL). Given the known association between S. aureus and squamous cell carcinoma (SCC), FCF shows the potential to effectively target and eradicate both SCC and the related S. aureus present within the lesions. In silico study reveals that Fagopyrin F effectively binds with the epidermal growth factor (EGFR), one among the highly expressed proteins in the A431 cells, with a binding energy of -9.6kcal/mol. The affinity of Fagopyrin F for EGFR on A431 cancer cells along with its cytotoxicity against skin cancer cells while safeguarding the normal cells (L929) plays a major part in the way it targets cancer cells. However, its safety, efficacy, and long-term advantages in treating skin conditions require more investigation, including in vivo investigations and clinical trials.

Biodiversity of ochratoxigenic Aspergillus species isolated from çavuş and karalahna grapes in Bozcaada, Türkiye

This study reported the presence and OTA production of ochratoxigenic Aspergillus species isolated from Bozcaada Çavuş and Karalahna grapes. The study was conducted with Aspergillus isolates isolated from grape samples collected during ripening in 2015 and 2016. Thin-layer chromatography was used to determine the secondary metabolite profiles of 290 Aspergillus isolates. Out of these, 122 isolates were found to be possible OTA producers. 43 isolates, selected based on their colony morphology of 122 isolates in different culture media, were identified using calmodulin gene sequencing analysis. The identified isolates were determined to be A. tubingensis, A. carbonarius, A. niger/welwitschia/awamori, A. welwitschia, A. spelaeus, and A. fructus. OTA production was investigated in six isolates. Using HPLC-FLD, these isolates were found to produce OTA in quantities ranging from 3.550 ± 0.240 to 92.346 ± 0.818 ppb. Consequently, OTA-producing Aspergillus species were isolated from grapes. The presence of A. spelaeus and A. fructus was reported for the first time in grapes. A. fructus has been found to be a new record for Türkiye.

Secondary metabolite fingerprinting, anti-pathogenic activity, elite chemotype selection and conservation of Curcuma caesia- an ethnomedicinally underutilized species.

Curcuma caesia Roxb. is an ethnomedicinally important, essential oil (EO) yielding aromatic plant. A total of twelve accessions of this plant rhizome were collected from six different agro-climatic zones of West Bengal, India and evaluated for their antimicrobial activities against eight disease-causing, multi-drug-resistant pathogenic strains of urinary-tract infection and respiratory-tract infection. The EO and extracts demonstrated antibacterial activity, with the highest inhibition zone of 18.00 ± 0.08 and 17.50 ± 0.14mm against Klebsiella pneumoniae by accession 06, even where all the broad-spectrum antibiotics failed to respond. In this study, we employed high-performance thin-layer chromatography (HPTLC) to quantify curcumin, the primary secondary metabolite of C. caesia, and the highest 0.228mg/gm of curcumin resulted from accession 06. Hence, on the basis of all aspects, accession 06 was identified as the elite chemotype among all twelve accessions. The chemical profiling of EO from accession 06 was done using gas chromatography-mass spectroscopy (GC-MS). Conceivably, about 13 medicinally significant compounds were detected. As this plant species is seasonal and has difficulties in conventional breeding due to dormancy, it must be conserved through in vitro tissue culture for a steady supply throughout the year in massive amounts for agricultural demand. A maximum number of 19.28 ± 0.37 shoots has been obtained in MS medium fortified with 6-Benzylaminopurine, Kinetin, and Naphthalene acetic acid. The genetic uniformity of the plants has been studied through Start Codon Targeted Polymorphism. Therefore, this study must help meet the need for essential phytoactive compounds through a simple, validated, and reproducible plant tissue culture protocol throughout the year.

Validation of HPLC and TLC analytical methods to determine radiochemical purity of 99mTc-cAbVCAM1-5, a new experimental radiotracer

Cardiovascular diseases, including fatal myocardial infarctions from atheromatous plaques, are the primary global mortality cause. Detecting stenotic atheromatous plaques is possible through coronary angiography, but vulnerable plaques with eccentric remodeling are undetectable with current diagnostic methods. Addressing this challenge, our group developed a radiopharmaceutical drug targeting vascular cell adhesion molecule 1 (VCAM-1), radiolabeled with technetium-99m. Given the absence of a monograph in the European Pharmacopoeia, and in order to draft the investigational medicinal product documentation, analytical methods had to be validated by high performance liquid chromatography (HPLC) and thin layer chromatography (TLC) to determine the radiochemical purity (RCP) of 99mTc-cAbVCAM1–5. This study therefore presents the results of the validation of analytical methods obtained in this context. The method validation followed the European Association of Nuclear Medicine (EANM) recommendations adapted from ICH Q2(R1), ensuring conformity with specificity, accuracy, repeatability and intermediate precision, linearity, robustness, quantification limit (LoQ), and range criteria. Regarding the results of specificity, both HPLC and TLC methods demonstrated excellent separation of 99mTc-cAbVCAM1–5 from impurities 99mTcO4-. Accuracy results indicated recovery percentages within the range of 99.52–101.40% for the HPLC and 99.51–101.97% for TLC, ensuring reliable measurements for each concentration of 99mTcO4-. Precision of the methods was validated by assessing repeatability and intermediate precision. Linearity was determined over the usual concentrations range and the correlation coefficient was greater than 0.99 for both methods. The limit of quantification was measured by diluting the 99mTcO4- to obtain a signal-to-noise ratio of around 10:1. Under these conditions, we obtained an LOQ of 2.10 MBq/mL for HPLC and 2Mbq/mL for TLC. In conclusion, the analytical methods developed in this study comply with EANM recommendations. This therefore allows us to correctly assess the radiochemical purity of 99mTc-cAbVCAM1–5, a new radiotracer targeting inflammation in vulnerable plaques.

Antiseizure Activity of Mitragyna inermis in the Pentylenetetrazol-Induced Seizure Model in Mice: Involvement of Flavonoids and Alkaloids.

This study aimed to investigate whether Mitragyna inermis (Willd.) Otto Kuntze organic and aqueous extracts are able to control seizures induced by pentylenetetrazol (PTZ) in mice based on flavonoid fingerprints and alkaloidal contents. Ethanolic extract and decoction-derived fractions from roots, leaves, and stems were subjected to chromatographic fingerprinting using AlCl3 and screening for their antiseizure effects using PTZ-induced acute seizure model. From the fractions that showed potent bioactivities, plausible antiseizure alkaloids were isolated using thin layer chromatography, and their structures were elucidated using 1H NMR, 2D NMR, 13C NMR, and FAB-HR (+ve or -ve). All fractions, with the exception of the dichloromethane and hexane fractions, revealed remarkable flavonoid fingerprints. An acute PTZ-induced seizure test revealed that ethanolic extract of stem bark [500 mg/kg body weight (bw)], ethyl acetate extract of stem bark (500 mg/kg bw), and aqueous extract of leaves (300 mg/kg bw) significantly delayed the occurrence of hind limb tonic extension (HLTE); however, a non-significant delay was observed in the onset of first myoclonic jerk compared with control animals. Isolation yielded four main alkaloids: that are, pteropodine (1), isopteropodine (2), mitraphylline (3) and corynoxeine (4). Corynoxeine is a new compound derived from M. inermis. This study suggests that flavonoid fingerprints are tracers of M. inermis anticonvulsant ingredients. The stem bark ethanolic and ethyl acetate extracts and leaf aqueous extracts contain anticonvulsant bioactive principles that delay notifying the HLTE occurring in male naval medical research institute mice. Furthermore, alkaloidal contents also remain plausible bioactive anticonvulsant principles. All observations support the traditional use of M. inermis to manage epilepsy. However, further studies are needed to understand the effects of alkaloid fractions, flavonoids, and the isolated compounds as promising antiseizure agents derived from M. inermis in experimental animals.