Thin-layer Chromatography Research Articles

Chitosan (CTS) induced secondary metabolite production in Canscora decussata Schult. - An endangered medicinal plant

Bioengineering has excellent potential in plant-based in vitro cultures for efficiently synthesizing desired plant products. Due to the rapid destruction of natural habitats, in vitro methods might save endangered species. Winged-Stem Canscora is an endangered medicinal herb used in traditional medicine for brain disorders. This study investigates the induction of in-vitro metabolite production using chitosan. Among the various concentrations of auxins tested, 82% of callus was obtained from Murashige and Skoog (MS) medium with 3 mg L−1 Kinetin (KIN) + 1 mg L−1 1-Naphthaleneacetic acid (NAA). The highest callus content was after the 3rd subculture at 200 mg L−1 chitosan treatment. On the 70th day of culture, levels of total phenolics and flavonoids are increased in the elicited and normal callus. However, antioxidant activity was higher in elicited callus compared to normal callus and wild plants. Higher secondary metabolite concentrations of elicited callus have superior anti-bacterial and anti-fungal activity. Secondary metabolites from elicited callus have a lower inhibitory concentration than those from wild plant and normal callus. High-performance thin layer chromatography (HPTLC) analysis showed that the elicited callus had higher amounts of mangiferin (0.178 μg mL−1) and scopoletin (0.133 μg mL−1). An efficient approach was employed to ensure that sufficient amounts of the widely used plant secondary metabolites from medicinal plants, such as mangiferin and scopoletin, were accessible to meet demand.

Exploring the chemogeographical variation of a commercially important medicinal tree (Prunus africana (Hook.f.) Kalkman) using a metabolomics approach

The stem-bark of Prunus africana (Hook.f.) Kalkman (African cherry) is traditionally used for the treatment of stomach ache, chest pain, malaria, fever, diabetes and high blood pressure. A large number of constituents have been isolated from the stem-bark including β-sitosterol and ursolic acid, which were reported to have anti-inflammatory activity. The aim of the study was to establish chemotypic variation among Prunus africana samples collected from three African countries, Cameroon, Democratic Republic of Congo (DRC) and Zimbabwe. Ninety-five stem-bark samples were collected from a total of nineteen populations, 11 in Cameroon (n = 55), 4 in Democratic Republic of Congo (n = 20) and 4 in Zimbabwe (n = 20). The samples were extracted with ethyl acetate, dichloromethane and methanol (1 g in 10 mL, 1 g in 10 mL and 2 mg in 5 µL, respectively). Chemometric analysis of chromatographic data obtained from high-performance thin layer chromatography (HPTLC), ultra performance liquid chromatography coupled with mass spectrometry (UPLC–MS), one-dimensional gas chromatography-time-of-flight-mass spectrometry (1D GC–ToF–MS) and spectroscopic data obtained from proton nuclear magnetic resonance (1H NMR) were done using MetaboAnalyst 4.0 software to explore chemotypic variation. A sample from each country was analysed by two-dimensional gas chromatography-time-of-flight-mass spectrometry (2D GC–ToF–MS) to resolve compounds that co-eluted in the 1D GC analysis. Quantification of selected compounds (β-sitosterol and ursolic acid) were done using a validated ultra performance liquid chromatography-photodiode-array detection (UPLC–PDA) method. The chemical profiles obtained from each of the four techniques were very similar for the samples from each country. The HPTLC profiles for Zimbabwe samples were distinctly different from the other two countries with a unique band at Rf value of 0.80. Multivariate analysis of the HPTLC data (using rTLC software), UPLC–MS, 1D GC–ToF–MS and 1H–NMR data revealed three clusters that were country specific. Chemometric analysis through the construction of PCA and a PLS–DA scores plots was performed, followed by the construction of variable important in projection plots, whereby marker compounds contributing significantly to the separation of the three clusters were selected and tentatively identified. Quantitative analysis revealed that samples from Zimbabwe contained higher levels of β-sitosterol (29.2 – 119 µg/g dry weight (DW) of the extract) compared to DRC and Cameroon, while samples from Cameroon were found to contain the highest levels of ursolic acid (1.80 – 141 µg/g DW of the extract). HPTLC, UPLC–MS, 1D and 2D GC–ToF–MS and 1H–NMR are valid tools for quality control of P. africana plant materials after revealing three clusters of samples collected from the three countries. Cameroon samples had high content of ursolic acid, while Zimbabwe samples showed high levels of β-sitosterol. The number of compounds detected across the three countries by 1D GC–ToF–MS was less compared to 2D GC–ToF–MS, which enhanced separation and resolution.

Thin layer chromatography assay to detect laccase inhibitors

Thin-layer chromatography (TLC) hyphenated to bioassays is a modern tool used for discovery of biologically active compounds from complex mixtures. The first bioautographic assay for detecting laccase inhibitors on a TLC plate was developed in this study. The on-plate reaction of laccase with colourless ABTS that renders the blue ABTS∙+ radical was optimised. Combination of the enzymatic TLC-assay with a control TLC-assay, wherein ABTS∙+ radical is chemically generated and then applied on the TLC, allowed to differentiate between the pure laccase inhibitor sodium azide and radical scavengers such as gallic and kojic acids. The limit of detection and quantification for the method were 54.9 and 166 ng of sodium azide respectively. The methodology was applied successfully to a recently discovered laccase inhibitor chemotype: hydrazones. A model hydrazone was compared with several hydrazones synthesized for this study. For the first time, laccase inhibitors separated on a TLC plate can be detected individually.

Pancreatic lipase inhibitory and anti‐acne activity of Piper betle: Kinetic studies, in silico docking, and chemical characterization of bioactive compounds

AbstractLipases play important roles in obesity and skin infections. Piper betle (Piperaceae) is widely cultivated and used in daily life. It is a rich source of lead compounds used in skin infections. This study reveals anti‐acne and pancreatic lipase inhibitory potential of leaf extract of Piper betle using in‐vitro and in‐silico molecular docking. The inhibitory potential of 13 selected plant extracts was evaluated at various concentrations (5–25 µg/mL). Ethyl‐acetate extract of Piper betle with strong pancreatic lipase inhibitory activity was further fractionated by preparative thin layer chromatography and eluted bands showing strong inhibitory activity were characterized by high performance liquid chromatography and gas chromatography. The purified compounds, namely, anodendroside E,2‐monoacetate, and 5H‐cyclopropa (3, 4) benz (1, 2‐ejazulen‐5‐one), were identified and showed over 90% pancreatic lipase inhibition. The isolated compounds showed strong dose‐dependent inhibitory effects on Propionibacterium acnes. These observations were consistent with molecular docking studies, which showed that anodendroside E,2‐monoacetate binds to the allosteric site with a binding energy of –7.0 kcal/mol, and this binding site is stabilized by hydrogen and carbon hydrogen bonds contributed by Phe 354 and Asp130, ser103. These findings suggest that the pancreatic lipase inhibitory and anti‐acne effects of Piper betle are attributed to anodendroside E,2‐monoacetate.

Characterization of carotenoids extracted from Haloferax larsenii NCIM 5678 isolated from Pachpadra salt lake, Rajasthan.

Carotenoids are a diverse group of pigments known for their broad range of biological functions and applications. This study delves into multifaceted potential of carotenoids extracted from Haloferax larsenii NCIM 5678 previously isolated from Pachpadra Salt Lake in Rajasthan, India. H. larsenii NCIM 5678 was able to grow up to OD600 1.77 ± 0.03 with carotenoid concentration, 3.3 ± 0.03µg/ml. The spectrophotometric analysis of carotenoid extract indicated the presence of three-fingered peak (460, 490 and 520nm) which is a characteristic feature of bacterioruberin and its derivatives. The bacterioruberin was purified using silica gel column chromatography and thin layer chromatography. The carotenoid extract showed 12.3 ± 0.09mm zone of growth inhibition with a minimum inhibitory concentration 546ng/ml against indicator strain, H. larsenii HA4. The percentage antioxidant activity of carotenoid was found to be 84% which was higher as compared to commercially available ascorbic acid (56.74%). Thus, carotenoid extract from H. larsenii NCIM 5678 possesses unique attributes with compelling evidence of antimicrobial and antioxidant potential for the development of novel pharmaceuticals and nutraceuticals.

Isolation and Identification of Isolate, S1 with High Biotransformation Potential of Ferulic Acid to Vanillin

Nine actinobacterial isolates were purified from the sediment sample of Kogilvai village, Warangal, Telangana, based on their capability to grow on the minimal medium with Ferulic acid (FA) as only Carbon (C) source. FA to Vanillin conversion capacity of these isolates was identified by Thin-layer chromatography (TLC). Biotransformation of FA to Vanillin was high by four isolates, S1, S3, O3 and O4 when compared to other five isolates (O1, O2, S2, S4 and S5) with initial pH 7 in basal medium. Among these four isolates, optimal and rapid FA to Vanillin bioconversion of 140 mg/L was shown by isolate S1 with UV-spectrophotometry. Its conversion was confirmed by High Performance Liquid Chromatography (HPLC) analysis with retention time of 2.9 min after 28hrs of incubation at 37°C with 1g/L ferulic acid in the medium with 150 rpm. Isolate S1 could utilize Lactose, Maltose, Glycerol, Fructose, Galactose, Sucrose, Dextrose, L-Arabinose, ONPG, Esculin and not other carbohydrates present in the Himedia Hicarbo kit. Molecular characterization showed that 16S rDNA gene sequence of isolate S1 was 98.27% similar to Limosilactobacillus fermentum CECT 562 with completeness of 96.7% and was identified as Limosilactobacillus sp. 16S rDNA gene sequence of isolate S1 was submitted to NCBI GenBank and its accession number was OR136396. As this isolate has high potential of FA to Vanillin biotransformation capacity, it can be further explored to be used for industrial setups for commercial exploitation.

Impact of ultra-high dilutions from Iranian endemic and commercial calendula on the germination and growth quality of Oryza sativa L.

PurposeThis article deals with producing ultra-diluted compounds (UHDs) prepared from Iranian calendula's characteristic and endemic species. It compares their chemical, biological and biochemical characteristics with the commercial sample of calendula species (grown in the Alps). In the following, these UHDs have been used to improve the quality of germination and growth and reduce contamination of rice (Oryza sativa) seeds in the laboratory environment. MethodsHigh-performance thin-layer chromatography (HPTLC) is used to isolate the active compounds. On the separated results, antioxidant and antibacterial were identified directly on the plate (Bio-autographic method). Direct on the plate)DESI mass spectrometry was used to identify the active compounds. ResultsThe HPTLC reveals that the chromatogram of native C. percica and C. officinalis extract is the most similar to the commercial compounds. The highest antioxidant activity is related to C. officinalis. The best antibacterial activity of the extracts against Staphylococcus aureus and Escherichia coli belongs to C. officinalis and C. tripterocarpa. Rutin, quercitrin, β-campstrole and di-o-caffeoylquinic acid, which are among the flavonoid and terpenoid categories were identified as active compounds. The prepared UHDs from native calendula are biologically more effective than the commercial ones in increasing seed germination efficiency, improving rooting quality and reducing contamination. ConclusionUsing UHDs increases the production of photosynthetic pigments the root length and the number of lateral roots. Also, the amount of protein, gibberellic acid and abscisic acid in seedlings treated using native UHDs of C. officinalis (native or commercial) is higher than the others.

Green Synthesis and Characterization of 4'-Hydroxy-4-Nitro Chalcone using Grinding Techniques

<p>The challenge of developing environmentally friendly synthesis methods for biologically active compounds remains a significant focus in green chemistry. This study aimed to synthesize 4'-hydroxy-4-nitro chalcone from 4-hydroxyacetophenone and 4-nitrobenzaldehyde using a grinding technique and then characterize the synthesized compounds. The 4'-hydroxy-4-nitro chalcone was synthesized via the Claisen-Schmidt grinding method for 45 minutes at room temperature. The grinding results were extracted with chloroform. The formed crystals were tested for purity by thin-layer chromatography (TLC), and further purification was carried out by recrystallization. The synthesized chalcone was characterized using <strong>FTIR,<sup> 1H</sup>-NMR</strong>, and <strong><sup>13</sup>C-NMR</strong> spectroscopy. The chalcone was also tested for antibacterial activity using the disc diffusion method against <em>Staphylococcus aureus</em> and <em>Escherichia coli</em>. The results showed that 4'-hydroxy-4-nitro chalcone could be successfully synthesized from 4-hydroxyacetophenone and 4-nitrobenzaldehyde using the grinding technique, yielding orangish-white crystals with a 70.63% yield and a melting point of 173°C. Purity was confirmed by TLC, and structural characterization was achieved through <strong>FTIR, <sup>1</sup>H-NMR</strong> and <strong><sup>13</sup>C-NMR</strong> spectroscopy. Antibacterial testing revealed that the synthesized chalcone produced a clear zone diameter of 9.27 mm against <em>S. aureus</em> and 27.88 mm against <em>E. coli</em>. These results indicate that the compound has relatively strong antibacterial activity against <em>E. coli</em> and relatively weak activity against <em>S. aureus</em>. This study demonstrates the effectiveness of the green synthesis method and provides valuable insights into the antibacterial properties of 4'-hydroxy-4-nitro chalcone</p>

Extraction, Isolation and Characterization of Bioactive Compounds from Euphorbia neriifolia (L.) Leaf and Evaluation of their Antioxidant Activity

Background: Amidst the identification of numerous secondary chemicals from Euphorbia neriifolia, there is a desperate need for the development of primary metabolite separation techniques. Objectives: In order to do that, bioactive chemicals from Euphorbia neriifolia (L.) leaf were extracted, isolated, and characterized. Subsequently, their antioxidant activity was evaluated. Methods: In this study, the determination of linoleic acid (LA) in petroleum ether extract (PEE) of Euphorbia neriifolia leaf (ENL) was carried out the first time by using thin-layer chromatography (TLC) and high-performance thin-layer chromatography (HPTLC) methods. Results: The chromatographic analysis of the PEE of ENL shows better spots and well-separated peak of LA with 0.49 retention factor (Rf) value and 22.54 ng LA content. The linearity of the calibration curve ranges from 5-25 ng/spot with a high correlation coefficient. The proposed method was characterized by better accuracy close to 99.5%, well robustness, and good precision range from 0.183% (intra-day) to 0.242% (inter-day). The percentage (%) RSD, which determined the stability of standard LA, did not exceed 2% after time period of 12, 24, 36, 48, and 72 h. The GC-MS analysis revealed the presence of different types of low or high-molecular-weight phytocompounds of varying quantities from the fractions of ENL. The FT-IR spectrum of ICs showed various peaks that confirmed the presence of C=C bending, C-H stretching, O-H stretching, CH2 stretching, and a carboxyl group. The 1H-NMR spectrum of the ICs from ENL confirmed the presence of octadecanoic in IC1, L-(+)-ascorbic acid dihexadecanoate in IC2, hexadecanoic acid in IC3, linoleic acid in IC4, and oleic acid in IC5, respectively. IC-4 showed greater antioxidant activity in comparison to other compounds with an IC50 value of 3.9 ± 0.01 μg mL-1. Conclusion: Thus, the present study identified five different phytocompounds that may be utilized as an effective option for the cure of different diseases.

Chemical analysis of Cyproheptadine Hydrochloride in appetite-stimulating herbs marketed in Banyumas regency: ensuring consumer safety

Jamu, a traditional Indonesian herbal medicine, is widely used in traditional healing practices nationwide, including the Banyumas Regency. The Indonesian government has implemented restrictions that forbid the inclusion of medicinal compounds in jamu, as their unregulated usage poses possible health hazards. Cyproheptadine hydrochloride is a frequently encountered medicinal component in appetite-stimulating herbs. However, the extent of adulteration of cyproheptadine hydrochloride in appetite-stimulating herbs marketed in Banyumas Regency is undisclosed. This study purposed to analyze and quantify the presence of cyproheptadine hydrochloride in appetite-stimulating herbs that are available for sale in the Banyumas Regency. The aim was to inform customers about these herbs' possible pharmaceutical ingredient content. The study utilized a qualitative analysis of thin-layer chromatography (TLC) and a quantitative analysis with a UV-Vis spectrophotometer. Fourteen samples of appetite-stimulating herbs in powdered, capsule, and tablet forms were gathered from different areas within the Banyumas Regency. Qualitative analysis revealed cyproheptadine hydrochloride in two herbal samples, SNM TG and SNM AM. The quantitative examination revealed that the content of cyproheptadine hydrochloride in SNM TG was 10.92 mg/5g, whereas in SNM AM, it was 7.93 mg/5g. These findings indicate that there have been breaches of regulations, and individuals who ingest these herbs may be susceptible to health problems. Further investigation is required to analyze a broader range of appetite-stimulating herbal samples, particularly those in liquid form, to ascertain the level of contamination with cyproheptadine hydrochloride and provide essential data for health regulations.

Metabolomic insights and bioactive efficacies of Tragopogon dubius root fractions: Antioxidant and antiproliferative assessments

Tragopogon dubius is commonly consumed as a vegetable and used in traditional medicine for treating inflammatory skin conditions and cutaneous swelling. Despite known pharmacological properties of its leaves and roots, many of its biological characteristics and active phytochemicals remain unexplored. The present study investigates the phytochemical composition, antioxidant, and anticancer properties of methanolic root extracts and isolated fractions (TdRM-1 and TdRM-2) of T. dubius. Utilizing preparative thin-layer chromatography, the crude extract was successfully separated into TdRM-1 and TdRM-2, characterized by GC-MS and FTIR analysis, revealing a diverse range of bioactive compounds including terpenes, flavonoids, and phenolic acids. Qualitative phytochemical screening indicated the presence of carbohydrates, tannins, alkaloids, and other phytoconstituents. Advanced UPLC-ESI-QTOF-MS analysis identified 54 metabolites, significantly contributing to the chemical profiling of the extract. The antioxidant activities of the fractions were quantitatively assessed using ABTS, DPPH, and superoxide radical scavenging assays, where TdRM-2 exhibited superior activity with IC50 values ranging from 51.29 to 60.03 μg/mL. Anticancer potential was evaluated against A549, LN-18, and MCF-7 cancer cell lines, demonstrating that TdRM-2 significantly inhibited cell proliferation with GI50 values as low as 31.62 μg/mL for A549 cells. Additionally, fluorescence microscopy revealed that TdRM-2 induces apoptosis, indicated by changes in nuclear morphology and loss of mitochondrial membrane potential. Annexin V-FITC/PI double staining indicate that the TdRM-2 fractions from T. dubius can significantly inhibit the growth of A-549, LN-18, and MCF-7 cancer cell lines by inducing apoptosis These findings suggest that T. dubius root extracts, particularly the TdRM-2 fraction, hold promising therapeutic potential due to their significant antioxidant and anticancer activities, underpinned by their rich phytochemical composition. This study underscores the importance of T. dubius as a source of natural bioactive compounds with potential health benefits.

Advances in Chromatographic Analysis of Phenolic Phytochemicals in Foods: Bridging Gaps and Exploring New Horizons.

Chromatographic analysis of phenolic phytochemicals in foods has significantly advanced over the past decade (2014-2024), meeting increasing demands for precision and efficiency. This review covers both conventional and advanced chromatographic techniques used for detecting phenolic phytochemicals in foods. Conventional methods like High-Performance Liquid Chromatography, Ultra High-Performance Liquid Chromatography, Thin-Layer Chromatography, and Gas Chromatography are discussed, along with their benefits and limitations. Advanced techniques, including Hydrophilic Interaction Liquid Chromatography, Nano-LC, Multidimensional Liquid Chromatography, and Capillary Electrophoresis, are highlighted for their innovations and improved capabilities. The review addresses challenges in current chromatographic methods, emphasizing the need for standardized and validated procedures according to the Food and Drug Administration, European Cooperation for Accreditation of Laboratories, and The International Organization for Standardization guidelines to ensure reliable and reproducible results. It also considers novel strategies for reducing the environmental impact of chromatographic methods, advocating for sustainable practices in analytical chemistry.

Bioconversion of Alternative Substrates for the Biosynthesis of HMG-CoA Reductase Inhibitors by Aspergillus spp. Strains with Antimicrobial Potential

Simvastatin, a semisynthetic drug widely used to lower cholesterol, is among the most prescribed statins worldwide. This study focuses on the direct production of a simvastatin-like biomolecule using alternative substrates by Aspergillus spp. strains. Two species, A. terreus UCP 1276 and A. flavus UCP 0316, were initially evaluated in synthetic media as control. Subsequently, the carbon and nitrogen sources were replaced by agro-industrial substrates, resulting in five modified media. Cultures were maintained at 28 °C, pH 6.5, at 180 rpm for 21 days. Fungal growth kinetics were evaluated and a 23 full-factorial design (FFD) was used to investigate the influence of substrate concentration on statin yield. Presence of inhibitors was confirmed by bioassay, UV–visible spectrophotometry, and thin-layer chromatography (TLC). According to the results, A. flavus UCP yielded 0.24 mg/g of statin in condition 2 of FFD (medium containing 4.5% soluble starch and saline base), suggesting it as a promising candidate for direct production of the biomolecule. Statistical analysis showed the significant effect of soluble starch on inhibitor production, making it a viable and profitable alternative substrate. Moreover, the isolated statin exhibited broad-spectrum antimicrobial activity, including efficacy against Gram-negative and Gram-positive bacteria and yeasts, indicating therapeutic potential against antimicrobial resistance.

Penalization and Color Code-Technical Approaches for Methods' Greenness and Whiteness Appraisal in Veterinary Medication: Assay of Toltrazuril Suspension.

Intestinal coccidiosis is a debilitating disease in poultry and livestock, leading to economic impact worldwide. Coccidiosis is prevented and treated in broilers by the inclusion of anticoccidials in feed. Toltrazuril is administered in potable water to treat coccidiosis. Three robust analytical methods for quantitation of toltrazuril in pure and pharmaceutical formulations are developed. Furthermore, ecological metrics; either penalization- or color-code-based techniques are applied for the appraisal of assays. Firstly, Second-Derivative (Δλ; 5 nm) spectrophotometric method; Toltrazuril is measured from peak to peak at 244-260 nm within a linearity range of 5-25 μg/mL. The second one is a high-performance thin-layer chromatography (HPTLC) analysis performed on an aluminum sheet of silica gel using ethyl acetate, methanol, ammonium chloride buffer, and water (8:1:0.5:0.5) (%V/V) as the elution phase. Toltrazuril, at a retardation factor of 0.66 ± 0.01, is linearly determined in the range of 1-9 μg/spot at 243 nm. The third one is Reversed Phase-HPLC-diode array detection, using Agilent column C18 (5 μm, 4.6 x 150 mm) in isocratic elution mode with a mobile phase of acetonitrile and water in a ratio of 80:20 (v/v), respectively, at 1 mL/min flow rate. Toltrazuril elutes at a retention time of 2.58 ± 0.1 min and is linearly determined at 243 nm in the range of 0.25-25 μg/mL. Calculated 2D-values and peak areas are highly correlated to their corresponding drug concentrations at coefficients; r > 0.999. All methods were ICH validated and applied to dosage form with satisfactory % recoveries (97-103%). Statistical comparisons reported one using t-test and F-test disclose insignificant variation. Examining greenness and whiteness norms, proposed methods were evaluated and ranked alongside four different reported methods. The proposed methods are green, accurate, and can be applied in routine quality control for the determination of toltrazuril in pharmaceutical formulations.

Does Trema micranthum (L.) Blume Produce Cannabinoids?

There are inconclusive claims in the scientific literature that the species Trema micranthum, widely distributed throughout the Brazilian territory, may produce phytocannabinoids, potentially serving as an alternative to Cannabis sativa. In this study, we conducted a comprehensive investigation to assess the presence of phytocannabinoids in two Trema micranthum samples collected in the Midwest region of Brazil. In trying to detect cannabinoids in T. micranthum, a recommended cannabis screening test was employed, the Fast Blue BB Salt (FBBBS) colorimetric assay, followed by thin-layer chromatography (TLC) and instrumental techniques: high-performance liquid chromatography coupled to diode array detector (HPLC-DAD) and gas chromatography coupled to mass spectrometry (GC-MS). When employed without chloroform extraction, the FBBBS reagent yielded positive results for extracts from all parts of T. micranthum (leaves, branches, fruits, and inflorescences). However, these initial positive results from the FBBBS test, suggesting the presence of cannabinoids, were not corroborated by FBBBS followed by chloroform extraction, TLC, or the instrumental techniques used in this study. These additional outcomes suggest that the positive FBBBS test results were likely due to the presence of other phenolic compounds rather than phytocannabinoids. For example, the presence of vitexin-like compounds in T. micranthum extracts might explain the positive FBBBS test results. Therefore, new assertions that T. micranthum produces cannabinoids will require the support of more selective experiments to avoid false-positive claims based on less selective screening tests.

Comparative Analysis of a Secondary Metabolite Profile from Leaves, Peel and Bulbs of Allium sativum L. by GC-MS

Garlic is a herbal plant that is widely used as a medicinal ingredient, because it contains secondary metabolite compounds such as alkaloids, flavonoids, tannins, phenols, terpenoids and saponins. The secondary metabolite content of plants is influenced by genetic, ontogenic, morphogenetic and environmental factors that can influence the biosynthesis and accumulation of secondary metabolites. Garlic is generally only used for the bulbs, while other parts such as the leaves and skin are still not widely used. This study aims to identify the secondary metabolite profile of ethanol extract of garlic leaves, bulbs and peels using the gas chromatography mass spectrometry (GC-MS) method. Identification of secondary metabolite compounds contained in each 70% ethanol extract was analyzed using the tube test, thin layer chromatography (TLC) and GC-MS methods. The results of analysis of secondary metabolite content using the tube test and TLC method showed that the ethanol extract of garlic leaves, skin and bulbs positively contained flavonoids, tannins, phenols and saponins. The results of secondary metabolite profile analysis using the GC-MS method from the ethanol extract of garlic skin and bulbs showed that 5 compounds were obtained with the main compound being 3,4-Dihydroxyphenylacetic acid and 23 compounds with the main compound being 1,2-Benzenedicaboxylic acid and palmitic acid, while the leek extract white only detected 1 compound. The conclusion is that the ethanol extract of garlic leaves, skin and bulbs has secondary metabolites in the form of flavonoids, tannins, phenols and saponins. There are differences in the profile of secondary metabolite compounds in the ethanol extract of garlic leaves, skin and bulbs in terms of the results of GC-MS analysis.

Empowering Future Pharmacists in Chemistry Analysis of Nutraceuticals

The community service aims to empower future pharmacists in East Java, Indonesia, by enhancing their understanding of nutraceuticals through chemistry analysis. Students gain essential skills for their academic pursuits through practical training in chromatography analysis and antioxidant activity measurements. The service equips students with practical skills in analyzing nutraceuticals, preparing them for further pharmacy studies. A training program was carried out for 88 SMA and SMK Farmasi students using Participatory Action Research (PAR). The main subjects covered were analysis using thin-layer chromatography, evaluation of functional groups in polyphenols, and assessment of antioxidant effectiveness. Pharmacy lecturers facilitated the program, guiding participants in practical competencies. The outcomes were successful, with a better understanding of nutraceuticals and valuable analytical skills gained. The significance of practical training and educational interventions in preparing students for advanced pharmacy studies is emphasized by this service.

Synthesis, In vivo, and In silico Evaluation of New Pyrazoline-Benzothiazole Conjugates as Antiepileptic Agents.

New 2-(4-benzothiazol-2-yl-phenoxy)-1-(3,5-diphenyl-4,5-dihydro-pyrazol-1-yl)-ethanones (9a-o) have been designed and synthesized. All the synthesized compounds were characterized by thin layer chromatography and spectral analysis. The antiepileptic potential of the synthesized compounds has been tested by following standard animal screening models, including maximal electroshock (MES) and subcutaneous pentylenetetrazole (scPTZ) models. The neurotoxic and antidepression effects of the synthesized compounds were checked by utilizing rotarod apparatus, and motor impairment test (by actophotometer) respectively. The study concluded that compounds 9c, 9d, 9f, 9i, 9n, and 9o possessed good antiepileptic potential compared to standard drugs like carbamazepine and phenytoin. The results of the rotarod performance test also established them without any neurotoxicity. The motor impairment test revealed that the synthesized compounds are also good antidepressants. In-silico studies have been performed for calculation of pharmacophore pattern, prediction of pharmacokinetic properties which determine the eligibility of synthesized compounds as orally administered molecules and interactions with the target proteins. The result of in-silico studies reinforced results obtained by in vivo study of the synthesized compounds and their possible mechanism of antiepileptic action i. e. via inhibiting voltage-gated sodium channels (VGSCs) and gamma-aminobutyric acid-A receptor.

Tissue-Specific Natural Synthesis of Galanthaminein Zephyranthes Species and Its Accumulation in Different In Vitro-Grown Organs Following Methyl Jasmonate Treatment.

Galanthamine is an immensely valuable alkaloid exhibiting anti-cancer and antiviral activity. The cultivation of plant tissues in in vitro conditions is a good source for the synthesis and enrichment of secondary metabolites of commercial interest. In this study, the Amaryllidaceae alkaloid galanthamine was quantified in three Zephyranthes species, such as Zephyranthes candida, Zephyranthes grandiflora, and Zephyranthes citrina, and the impact of the methyl jasmonate (MJ) signaling molecule on galanthamine accumulation was monitored in in vitro-derived plant tissues. This is the first ever study of the MJ-regulated accumulation of galanthamine in in vitro-grown Zephyranthes tissues. Shoot regeneration was obtained in all three Zephyranthes species on Murashige and Skoog (MS) medium containing 2.0 mgL-1 benzylaminopurine (BAP) + 0.5 mgL-1 naphthalene acetic acid (NAA). The regenerated shoots were rooted on a medium containing 2.0 mgL-1 indole butyric acid (IBA). A GC-MS study of Zephyranthes extracts revealed the presence of 34 phyto-compounds of varied levels with therapeutic activities against diseases. The galanthamine content was quantified in plant parts of the three Zephyranthes species using high-performance thin layer chromatography (HPTLC); the maximum was found in Z. candida bulb (2.41 µg g-1 dry wt.), followed by Z. grandiflora (2.13 µg g-1 dry wt.), and then Z. citrina (2.02 µg g-1 dry wt.). The galanthamine content showed bulb > leaf > root source order. The in vitro-generated plantlets were treated with different MJ concentrations, and the galanthamine yield was measured in bulb, leaf, and root tissues. The highest galanthamine content was recorded in bulbs of Z. candida (3.97 µg g-1 dry wt.) treated with 150 µM MJ, showing an increase of 64.73% compared to the control. This accumulation may be attributed to MJ-induced stress, highlighting the potential commercial synthesis of galanthamine in vitro.

Progress, applications, and challenges in high-throughput effect-directed analysis for toxicity driver identification - is it time for HT-EDA?

The rapid increase in the production and global use of chemicals and their mixtures has raised concerns about their potential impact on human and environmental health. With advances in analytical techniques, in particular, high-resolution mass spectrometry (HRMS), thousands of compounds and transformation products with potential adverse effects can now be detected in environmental samples. However, identifying and prioritizing the toxicity drivers among these compounds remain a significant challenge. Effect-directed analysis (EDA) emerged as an important tool to address this challenge, combining biotesting, sample fractionation, and chemical analysis to unravel toxicity drivers in complex mixtures. Traditional EDA workflows are labor-intensive and time-consuming, hindering large-scale applications. The concept of high-throughput (HT) EDA has recently gained traction as a means of accelerating these workflows. Key features of HT-EDA include the combination of microfractionation and downscaled bioassays, automation of sample preparation and biotesting, and efficient data processing workflows supported by novel computational tools. In addition to microplate-based fractionation, high-performance thin-layer chromatography (HPTLC) offers an interesting alternative to HPLC in HT-EDA. This review provides an updated perspective on the state-of-the-art in HT-EDA, and novel methods/tools that can be incorporated into HT-EDA workflows. It also discusses recent studies on HT-EDA, HT bioassays, and computational prioritization tools, along with considerations regarding HPTLC. By identifying current gaps in HT-EDA and proposing new approaches to overcome them, this review aims to bring HT-EDA a step closer to monitoring applications.