Thin Layer Chromatography Analysis Research Articles

Mechanistic insight into neuroprotective effect of standardized ginger chemo varieties from Manipur, India in scopolamine induced learning and memory impaired mice.

Alzheimer's disease is a complex neurodegenerative disease characterized by progressive decline in cognitive function and behaviour. Ginger is the rhizome of the plant Zingiber officinale Roscoe, has been an important ingredient of many Ayurveda formulations to treat neurological disorders. The present study aims to estimate the variation of 6-gingerol content in nine different ginger samples collected from Manipur, India, investigate the neuroprotective potential of the most potent ginger sample against scopolamine-induced cognitively impaired mice, and validate the therapeutic claim by molecular docking analysis. High Performance Thin Layer Chromatography (HPTLC) analysis suggested that the sample GV6 had the highest 6-gingerol content with potent in vitro acetylcholnesterase (AChE) (IC50 = 336.10µg/mL) and butyrylcholinesterase (BChE) (IC50 = 411.73µg/mL) enzyme inhibitory activity. The neuroprotective potential of GV6 was tested in scopolamine-induced cognitively impaired mice (200 and 400mg/kg). The behavioral analysis showed that GV6 alleviated the spatial recognition, and short-term and long-term memory in the experimental mice model. GV6 significantly improved brain AChE and BChE activity, acetylcholine (ACh) level, markedly alleviated the antioxidant parameters, and reversed the neuroinflammation. Brain histopathological observations confirmed the presence of organized nerve fibers, improvement of neuronal cell density, and reverse the nucleus shrinkage. Further molecular docking analysis showed that 6-gingerol and galantamine exhibited stable interaction with AChE (-7.5 and - 7.3 kcaL/moL) and BChE (-7.3 and - 8.5 kcaL/moL). The present study emphasizes the quality-related therapeutic importance of ginger samples from Northeast India and demonstrates that administration of GV6 may improve brain cognitive functions by restoring neurotransmitter levels and inflammatory and antioxidant parameters in scopolamine-induced cognitively impaired mice.

Isolation and Antioxidant Mechanism of Polyphenols from Sanghuangporous vaninii.

Sanghuangporous vaninii, as an edible and medicinal macrofungus, represents a high source of polyphenols with considerable antioxidant activities. However, due to the significant differences in polyphenol content and bioactivity caused by different cultivation substrates, its antioxidant mechanism has not been fully determined. In this paper, five groups of S. vaninii fruiting bodies were collected from cultivation substrates from different areas. The ethanol extracts of mulberry sawdust from Haining City (HNMS) had the highest polyphenol content, as well as excellent antioxidant activity. HNMS3, a polyphenol component with promising antioxidant capacity, was further isolated through optimization with different extractants, silica gel column chromatography, and thin layer chromatography analysis. UPLC-Q-TOF-MS analysis showed that HNMS3 was composed of 33 compounds, corresponding to 257 targets of oxidative stress by network pharmacology analysis, which were strongly associated with mental health and neurodegenerative diseases. Protein-protein interaction and molecular docking analysis indicated that eight hub genes (PPARG, IL-6, STAT3, PTGS2, SRC, MTOR, ERS1, and EGFR) are attributed to the regulation of the key compounds hispidin, inoscavin A, inoscavin_C, and phellibaumin B. Consequently, this study obtains S. vaninii polyphenolic component HNMS3 with excellent antioxidant capacity, simultaneously revealing its potential antioxidant mechanisms, providing new insights into the application of S. vaninii.

Chromatographic’s image analysis in phytohomeopathies

Plants signal when establishing interaction with homeopathies in different succussions, however, the type of signaling is not always understandable. Recognizing symptomatic patterns in plants begins by understanding the behavior of homeopathy itself, through varied methods, widely disseminated in scientific literature¹. The present study proposes a new thin-layer chromatography (TLC) analysis method to identify patterns in different phytohomeopathic applications. CCDs were made on Sulfur 9Ch, Nux and Bell 11CH; Calc Ph 29CH; Bry and Cal Carb 30CH; Hyp 50CH; Mag Ph 200CH and Graph Nat 200CH. The analysis of the chromatographic stains was developed with ultraviolet light and the images were analyzed using the Fast Fourier Transform (FFT) method². The statistical design, ANOVA, consisted of 5 completely randomized, double-blind replications for each of the preparations. The results showed significant differences after a few minutes of chromatography, highlighting the order of signals between homeopathies: Bell < Sulph < Mag Ph < Graph Nat < Hyp < Bry < Calc Ph < Cal Carb < Nux. From these results, it is possible to identify phytohomeopathic’ signs and symptoms during the course of the treatment.

An improved matrix deposition technique for thin layer chromatography coupled to MALDI-TOF mass spectrometry (TLC-MALDI).

Analytical thin layer chromatography (TLC) is a simple yet powerful chromatographic technique that is widely used for the qualitative characterization of complex mixtures such as plant extracts. For their qualitative and visual characterisation, a large number of more or less specific colour reactions are at hand and numerous reference substances are available as well. However, the identification of extract components by colour and the comparison of retention times is not straightforward. In contrast, the coupling of TLC with MALDI-TOF mass spectrometry can enable the identification of components and contribute to the optimization of TLC protocols. One of the most important steps for a successful TLC-MALDI process is the deposition of a sufficient amount of matrix onto the TLC plate. Standard methods such as the dip-coating protocol have major drawbacks. Here we present an improved and robust procedure for matrix application by means of matrix lines. The practicability of the method was tested on plant extracts from Agrostemma githago L. and Papaver somniferum L. (opium).

Phytochemical analysis and quantification of flavonoid content of ethyl acetate fraction from ‘Alugbati’ (Basella rubra L.) leaves

Basella rubra L. (Alugbati) is a branching, herbaceous vine with heart-shaped leaves widely grown in Asia, with the leaves used as a herb in soups and salads. Several studies have highlighted this plant’s nutritional and biological benefits, which contain various phytochemical substances, including flavonoids. Flavonoids are polyphenolic compounds known to scavenge free radicals through specific mechanisms. This study aimed to extract and quantify flavonoids from the leaves of Basella rubra L. to emphasize its potential benefits as a food or nutraceutical. The dried leaves were pulverized and macerated with 99% methanol before partitioning with ethyl acetate and water. The fractions were tested qualitatively and then analyzed using Fourier Transform Infrared Spectroscopy (FTIR) to detect the presence of flavonoids, followed by thin layer chromatographic (TLC) analysis to confirm the presence of flavonoids. A total flavonoid content (TFC) test was performed to quantify the extracted flavonoids. The positive results from the qualitative tests and the subsequent FTIR spectra reflecting the constituent bonds and functional groups validated the presence of flavonoids in the plant fractions. The aqueous and ethyl acetate fractions yielded a total flavonoid content of 2.71 ± 0.01 and 9.61 ± 0.04 mg QE/g extract, respectively. The plant's physical and environmental background, the extraction process used, and the plant part used are all potential contributors to its flavonoid content. To maximize its nutritional and therapeutic potential, further research into other plant organs, and performing other extraction and purifying procedures, is recommended.

Comparative Study of Antimicrobial Efficacy and Phytochemical Profiles of Ethanol and n-Hexane Extracts from the Roots of Ixora coccinea Linn

The root extracts of Ixora coccinea were evaluated for their phytochemical composition, antibacterial activity, and minimum inhibitory concentration (MIC) against Escherichia coli, Staphylococcus aureus, and Bacillus subtilis. Quantitative phytochemical analysis was conducted to determine the levels of terpenoids, flavonoids, alkaloids, tannins, and saponins. Thin Layer Chromatography (TLC) was used to confirm the presence of bioactive components in the extracts under different solvent systems, followed by antibacterial susceptibility tests using the disc diffusion method. The MIC was determined through serial dilution to identify the lowest concentration that inhibited bacterial growth. Quantitative phytochemical analysis showed that the ethanol extract contained 14.80% flavonoids, 6.00% alkaloids, 1.20% saponins, 1.80% terpenoids, and 0.03% tannins. In contrast, the n-hexane extract had higher terpenoid content (3.40%) but lower flavonoids (5.20%) and alkaloids (2.10%). Thin Layer Chromatography (TLC) analysis under varying solvent systems showed different Rf values, with the ethanol extract displaying more diverse spots, indicating a richer variety of bioactive compounds. Antibacterial susceptibility tests revealed that the ethanol extract produced larger inhibition zones (10 ± 0.4–28 ± 1.0 mm) than the n-hexane extract (6 ± 0.2–26 ± 0.8 mm). MIC results indicated that ethanol extract inhibited E. coli and S. aureus at 50 mg/L and B. subtilis at 25 mg/L, while the n-hexane extract required 100 mg/L for effective inhibition. In conclusion, the findings indicate that the plant possesses significant antibacterial potential, warranting further research to isolate and characterize the specific phytoconstituents responsible for the observed antimicrobial activities.

ISOLATION AND CHARACTERIZATION OF WATER SOLUBLE FRACTION OF PROPOLIS AND ITS ANTIBACTERIAL POTENTIAL ON BACTERIA CAUSING CONJUNCTIVITIS

Propolis is a bee product with a variety of biological activities. Although chemical composition of propolis differs by the location but all propolis types possesses antimicrobial activity. The usage of propolis for apitherapeutic purposes has increased recently. But its ethanol solubility limits its usage in certain areas like ophthalmology. Main objective of this study is to isolate water soluble components of propolis and determination of its antimicrobial activity against two bacteria causing conjunctivitis namely Neisseria gonorrhoeae and Haemophilus influenzae. Isolation of water soluble fraction of propolis was carried out in two steps by using pectin-propolis micro beads. Isolated water soluble fraction and crude extract was examined by thin layer chromatography and HPLC analyses. Three main spots were screened on TLC plate after isolation. These spots could be explained by the presence of different class of compounds in the isolate. HPLC analyses showed that water soluble fraction contained phenolic acids, their esters and flavonoids like p-OH benzoic acid, t-cinnamic acid, pinocembrin and caffeic acid phenethyl ester. Low antimicrobial activity was achieved against tested microorganisms for the fraction. It can be concluded that isolation of water soluble fraction of the propolis extract could be a solution for its usage in restricted areas.

GC-MS/HPLC Profiling and Sono-Maceration Mediated Extraction of Osbeckia Parvifolia Polyphenols: In Silico and In Vitro Analysis on Anti-Proliferative Activity in Ovarian Cancer Cell Lines.

Osbeckia parvifolia, an endemic edible plant of Western Ghats, was investigated in the present study for its polyphenolic compounds, including content, constituents, extraction through an ultrasonic-assisted maceration technique and therapeutic potential in biomedical applications. The methanolic extract (OPM) exhibited an IC50 value of 1.25 μg/mL against 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radicals. Furthermore, the ethyl acetate and methanolic extracts also strongly inhibited 5-lipoxygenase, especially OPM (84.93 %), which was comparable to standard curcumin. OPM also elicited cytotoxicity in SKOV3 ovarian cancer cells (93.80 %), surpassing paclitaxel. Bio-accessibility analysis demonstrated that the release of phenolic compounds and antioxidant potential were very high (above 100 %), revealing the possibility of synergistic efficacy of polyphenolic complexes in drug development. Gas Chromatography -Mass Spectrometry (GC-MS) analysis revealed 22 bioactive polyphenolic compounds in OPM, such as epicatechin, quercetin, and psoralidin. This was confirmed by High Performance Liquid Chromatography (HPLC) and High-Pressure Thin Layer Chromatography (HPTLC) analyses, which revealed a high quantity of catechin (37.45 mg/g). Molecular docking revealed the significant binding affinity of these proteins for the ovarian oncoproteins PI3K (-8.52 kcal/mol) and Casp-8 (-8.41 kcal/mol). Adsorption, Distribution, Metabolism, Excretion and Toxicity (ADMET) profiling indicated the favorable pharmacokinetic properties of these compounds, supporting their candidacy in drug formulations against ovarian cancer.

Deciphering supramolecular arrangements, micellization patterns, and antimicrobial potential of bacterial rhamnolipids under extreme treatments of temperature and electrolyte.

The micellization properties of rhamnolipids (RLs) in extreme electrolyte concentrations and temperatures have gained considerable attention due to their broad industrial applications. In this study, the aggregation behavior, specifically the micellization pattern (critical micelle concentration (CMC)) of RLs produced from a newly isolated thermophilic strain of Pseudomonas aeruginosa from a harsh environment of an oil field, was investigated by a spectrophotometric method at various temperatures (293-393 K) and electrolyte concentrations (NaCl: 2-20%). The result indicated that the CMC values (0.267-0.140 mM⋅dm-3) were both electrolyte- and temperature-dependent exhibiting a U-shaped trend as temperature and NaCl concentration increased. Variations in NaCl concentration and temperature also affected the standard Gibbs free energy (ΔGo mic ), enthalpy (ΔHo mic ), and entropy (ΔSo mic ) of micellization. The molecule also showed stability at a broad range of temperatures, pH, and NaCl concentrations. Thin-layer chromatography (TLC) and Fourier-transform infrared (FTIR) analysis confirmed the similarity in composition between the crude extract and the commercial RL with Rf values of 0.72 for mono-rhamnolipids and 0.28 for di-rhamnolipids. FTIR analysis confirmed the chemical nature particularly key aliphatic functional groups present in the fatty acid tail of RLs and the -COC- bond in the structure of the rhamnose moiety. Additionally, LC-ESI-QTOF analysis confirmed corresponding ionic fragments of mono- and di-rhamnolipids congeners. Furthermore, the antimicrobial potential was determined against different human pathogens in the absence and presence of NaCl by measuring zones of inhibition. The result revealed enhanced inhibitory effects against Gram-positive pathogens (S. aureus, S. epidermidis, and L. monocytogene), with zones of inhibition of 26, 30, and 20 mm in the presence of NaCl. These findings underline the role of NaCl in the micellization of RL molecules and highlight their importance in environmental applications, pharmaceuticals, and various life science sectors.

Preservation of freshly-cut lemon slices using alginate-based coating functionalized with antioxidant enzymatically hydrolyzed rice straw-hemicellulose

Food coatings are efficient preservative measures, a crucially needed approach to meet hunger growth as well as food management. In the current study, the construction of an efficient coating using alginate polymer fortified with antioxidant rice straw-hemicellulose hydrolysate was examined. Rice straw hemicellulose fraction was extracted under thermal alkaline conditions with a recovery percentage of 15.8%. The extracted hemicellulose fraction was enzymatically hydrolyzed with microbial xylanase with hydrolysis percentage of 53.8%. Characterization of the produced hydrolysate was performed with the aid of thin layer chromatographic analysis (TLC), high-performance liquid chromatographic analysis (HPLC), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) analysis. The reported data showed that xylobiose (240.68 mg/g) in addition to coumaric (383.33 µg/g) and ferulic acid (298.77 µg/g) as the main constituents of the carbohydrate and the polyphenolic contents, respectively. The hydrolysate possessed antioxidant capacity that significantly increased in a direct correlation with the concentration of the hydrolysate. Finally, the prepared coating solution effectiveness in the preservation of lemon slices against fungal growth was monitored up to 20 days with a significant concentration dependent decrease in weight loss and an increase in its antioxidant activity. The combination of xylooligosaccharide-rich rice straw hydrolysate with alginate-based coating not only improved the storage shelf-life of fresh fruits and vegetables but also provided microbial safety and potential benefits for human health.

Antibacterial Properties in Fermentate Extracts of Endophytic Fungus Derived from Marine Algae Caulerpa lentillifera Against Gastrointestinal Pathogenic Bacteria

Endophytic fungi, residing symbiotically within living plant tissues, are a promising source of novel bioactive compounds. These fungi are known to produce secondary metabolites with a wide range of biological activities, including antibacterial properties. This study explored the antibacterial potential of fermented extracts from endophytic fungi isolated from the marine alga Caulerpa lentillifera, a food source consumed by coastal communities and traditionally used for medicinal purposes. Thin-layer chromatography (TLC) bioautography was employed to identify extracts with activity against common gastrointestinal pathogenic bacteria, such as Escherichia coli, Shigella dysenteriae, and Salmonella typhi. Two endophytic fungal isolates displayed promising antibacterial activity against these pathogens. TLC analysis tentatively identified the presence of flavonoids and alkaloids, potentially responsible for the observed effects. This research sheds light on the pharmacological potential of endophytic fungi and their applications in combating bacterial infections.

High-Performance Thin Layer Chromatography (HPTLC) Analysis for the Quantification of Constituents in Ficus religiosa

High-Performance Thin Layer Chromatography (HPTLC) Analysis for the Quantification of Constituents in Ficus religiosa

Pharmacognostical and phytochemical studies of Eulophia dabia (D. Don) Hochr: A folklore medicine

Abstract BACKGROUND: Medicinal plants have been used in traditional systems of medicine since ancient times. In India, many plants use the common name “Salam mishri,” which creates huge controversy and confusion among the plants. But among them, Dactylorhiza incarnata subsp. Incarnate, Eulophia dabia (E. dabia) (D. Don) Hochr. and Dactylorhiza hatagirea (D. Don) are considered Salam mishri by several local ethnic groups and villagers, as they exhibit the same medicinal properties. E. dabia (D. Don) Hochr. is also known by several vernacular names such as Munjatak (Sanskrit), Salam mishri (Hindi), etc. It is extensively used traditionally to enhance the body’s strength and ability. METHODS: Various methods, including microscopic, macroscopic, and physico-phytochemical methods, were applied to determine the diagnostic features for the standardization and identification of E. dabia using the guidelines for drug development of Ayurved formulations given by CCRAS. RESULTS: Preliminary physicochemical analysis revealed a loss on drying of 4.84% w/w, pH of 5.16, alcohol-soluble extractive value of 43.56% w/w), and water-soluble extractive value of 63.05% w/w. The total ash content was 6.3% w/w, with acid-insoluble ash at 2.03% w/w for the rhizome of E. dabia. Powder microscopy of the rhizome revealed the presence of stone cells, starch grains, parenchyma cells, cork cells, and xylem vessels. Thin-layer chromatography and high-performance thin-layer chromatography analysis showed the presence of important phytoconstituents in the methanolic extract of the rhizome. CONCLUSION: The data generated from this study would be helpful in the authentication of this medicinal plant. These pharmacognostic and preliminary phytochemical observations can be considered future references in further studies.

Maximization of Biodiesel production from oil that produced during salmon smoking process with high amount of omega-3 by using homogenous catalyst.

The escalating global demand for oil, coupled with declining fossil fuel production, prompts the urgent exploration of renewable alternatives. To address this challenge, researchers are actively seeking environmentally friendly fuels like biodiesel. Among potential feedstocks, oil that is produced from salmon smoking process during industry emerges as a promising option. Smoked salmon oil could be a challenge when producing biodiesel due to its high content of omega-3 compounds. Using homogenous commercial catalyst from alkali to achieve the highest yield from salmon smoking oil was the aim of the current study. A Box-Behnken design and response surface methodology in Design Expert software (version 13) was used to study the effect of four main factors on the biodiesel yield from salmon smoking oil. The optimum biodiesel values were 70°C, 90 min, 0.753 wt.%, and 20 wt.% for temperature, reaction time, sodium hydroxide concentration, and methanol concentration, respectively. At these optimum values, the highest biodiesel production was 92.0% with fatty acid methyl ester contents of 83.4% and conversion efficiency of 77%. Thin-layer chromatography and thermal gravimetric analysis confirmed the successful production of biodiesel at optimized conditions. Using Aspen Plus simulation software confirmed the cost-effectiveness of the homogenous catalyst used for enhancing biodiesel production from salmon smoking oil.

Analysis of sildenafil citrate in male stamina herbal medicines from the Magelang region, Central Java using thin-layer chromatography and UV spectrophotometry

Background: Male stamina herbal medicines are widely consumed in Indonesia, often believed to enhance sexual performance. However, the illegal addition of medicinal chemicals (BKOs) such as sildenafil citrate in traditional herbal products poses health risks. Objective: To identify and quantify sildenafil citrate in selected herbal medicine samples using thin-layer chromatography (TLC) and UV spectrophotometry. Method: Three samples of male stamina herbal medicine were purchased and subjected to organoleptic tests. Ethanol (96%) was used for extraction, followed by TLC analysis using two mobile phases (methanol: chloroform at 4:1 and 1:1 ratios). UV spectrophotometry was employed to determine the maximum absorbance wavelength and quantify sildenafil citrate. Results: TLC analysis revealed that two samples (K and L) contained sildenafil citrate, with Rf values consistent with the standard. UV spectrophotometry confirmed the presence of sildenafil citrate at a maximum wavelength of 292.5 nm. Sample K contained 0.96%, while sample L had 13.766% sildenafil citrate. Conclusion: The presence of sildenafil citrate in two samples violates Indonesian regulations prohibiting medicinal chemicals in herbal products, raising significant safety concerns.

Phytochemistry of Petroselinum crispum (Mill.) Fuss, Murraya koenigii (L.) Spreng. and Cinnamomum tamala (Buch.-Ham.) T. Nees & C. H. Eberm. and in silico studies of the role of their bioactive components against cancer

Phytochemical profiling using reliable equipment and validated methods helps us know the medicinal value of the plants. Since natural compounds have fewer side effects, they can serve as replacements for synthetic drugs that are being used in the treatment of challenging chronic diseases like cancer. The present study focuses on the bioactive phytochemical profiling of Petroselinum crispum (Mill.) Fuss, Murraya koenigii (L.) Spreng., and Cinnamomum tamala (Buch.-Ham.) T. Nees & C. H. Eberm. and in silico studies to check the anticancer potential of their bioactive components. In thin-layer chromatography (TLC) plate analysis, it was found that the methanolic extracts of plants contained the maximum number of components. Gas chromatography–mass spectrometry (GC-MS) analysis of the methanolic extracts of plants showed the presence of 22 bioactive components. High-performance thin-layer chromatography (HPTLC) analysis of the extracts of these plants showed the important chromatographic peak, apigenin. In silico studies showed the binding efficacy of selected bioactive components observed in the analysis of plant extracts. Amongst them, apigenin was found to be most effective at binding to the receptors of targeted cancer cells, viz., hepatocellular carcinoma, lung, and breast cancer. After the analysis of the study, it was arrived at the conclusion that the plants, viz., P. crispum, M. koenigii, and C. tamala, possess various bioactive components, and some of these components have anticancer potential. Therefore, in vivo and in vitro studies should be essentially conducted for the development of cancer-preventive drugs.

Exploring novel antioxidant cyclic peptides in corn protein hydrolysate: Preparation, identification and molecular docking analysis

Antioxidant cyclic peptides were successfully identified from a corn protein hydrolysate. Hydrolysate by Alcalase + Flavourzyme showed the highest cyclic peptide purity (48.36 ± 1.81 %) and higher antioxidant activities compared with other hydrolysate. The success of peptide cyclization in hydrolysate was demonstrated by thermogravimetric analysis and thin-layer chromatography (TLC) analysis. Thermogravimetric analysis showed that the thermal stability of hydrolysate after cyclization was significantly increased, which was related to the formation of cyclic peptides. Peptides with molecular weight less than 1000 Da accounted for more than 80 % in hydrolysate after cyclization. After separation using gel silica chromatography and semi-preparative reverse phase high performance liquid chromatography (RP-HPLC), 22 novel antioxidant cyclic peptides were identified by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS) and orbitrap-tandem mass spectrometry (Orbitrap-MS/MS). Synthetic cyclic peptides with the same sequence were synthesized and characterized for their antioxidant activity. Molecular docking suggested that the free radical molecules could bind with the cyclic backbone and side chain of cyclic peptides through hydrogen bonding, hydrophobic interaction as well as electrostatic interaction. This study has important implications for the high-value utilization of corn protein and new cyclic peptides drugs or functional food development.

Unveiling the Potential of Hyptis Capitata and Hyptis Brevipes Compounds in Overcoming Apoptosis Resistance and Inducing Targeted Cell Cycle Arrest in Breast Cancer Stem Cells

Breast cancer stem cells (CSC), as a kind of tumor cells, are able to regenerate themselves, leading to apoptosis resistance and cancer relapse. In this study, we investigated the potential of Hyptis Capitata and Hyptis Brevipes compounds in preventing apoptosis resistance and inducing targeted cell cycle arrest in CSC. The cytotoxic effects of the compounds were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay and the cell cycle distribution and apoptosis induction were assessed through flow cytometry analysis. The phytochemical characterization of Hyptis bravipes extract (HBE) and Hyptis capitata extract (HCE) conducted under thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analysis. The results revealed that Hyptis Brevipes extract (HBE) exhibited strong cytotoxicity towards CSCs, with an IC50 value of 89 µg/mL. In contrast, Hyptis Capitata extract (HCE) showed weak growth inhibition, with an IC50 value greater than 500 µg/mL. Subsequent analysis demonstrated that HBE induced cell cycle arrest in the G2/M and S phases and significantly increased apoptosis in the CSC population. Furthermore, phytochemical characterization using TLC and HPLC revealed that HBE and HCE share similar constituent compounds, with HBE exhibiting a more diverse compound content and stronger cytotoxic effects. These findings provide valuable insights into the potential of Hyptis Capitata and Hyptis Brevipes compounds as targeted agents against breast cancer stem cells and contribute to understanding their chemical profiles and pharmacological properties. HIGHLIGHTS Hyptis brevipes extract induced cell cycle arrest in G2/M and S phases. Cytotoxicity of Hyptis brevipes leading to apoptosis induction on CSC Hyptis brevipes significantly targeted Breast Cancer Stem Cells GRAPHICAL ABSTRACT

GC-MS Analysis of Antibiotic-Producing Bacteria Isolated from The Soil Environment of Zamfara State

Antibiotics produced by soil microorganisms, are crucial for treating bacterial and fungal diseases. Due to antibiotic resistance, a broad spectrum of antibiotics from bacteria is needed. Soil samples from Zamfara state were used for detecting antibiotic-producing bacteria. The study involved physico-chemical analysis, pour plate methods, biochemical tests and molecular characterization using 16s rRNA. Agar well diffusion method was employed to test isolates’ ability to inhibit pathogens growth and their potential for antibiotic production. Antibiotics production was assessed through small-scale submerged fermentation condition. Compounds were extracted using solvent extraction method, thin layer chromatography and GC-MS analysis were used for separation and identification. Data analysis were done using SPSS and were presented as mean, standard error of mean, and Anova. The soil samples exhibited light brown to dark brown colors, indicating clay-loamy, loamy-sandy and sandy-loamy. Sample C had the highest pH (7.31±.03), Sample H had the highest temperature (26oC), sample D had the highest electrical conductivity (108µs/m), and sample B had the highest moisture content (5.6). Six bacteria were identified and these include; Bacillus cereus, Bacillus pumilus, Providencia stuartii, Providencia alcalificiens, Pseudomonas aeruginosa, and Micrococcus luteus. All the organisms inhibited at least one pathogen, with Bacillus cereus showing the highest zone of inhibition. GC-MS analysis revealed diverse bioactive compounds namely, Carbonic acid, Glycerin, Nonadecane, 5-hydroxymethyl furfural, L-arabinitol, and Methylchloroformate. This study has demonstrated the diversity of bacterial population in the local soils and potential production of antibiotics that can be useful for many medical applications and further studies are recommended.

Isolation and Identification of Bioactive Chlorogenic Acid from Ethanol Fraction of Cordia subcordata: HPLC, HPTLC, FTIR, and LC-MS Analysis.

Cordia subcordata was subjected for soxhlet extraction using soxhlet apparatus by hydroalcoholic solvent (Ethanol:water; 70:30). The obtained crude extract was subjected to organoleptic properties. The extract was dark greenish or greenish-black in color. The odor of the extract was strong like tamarind. The extract taste was bitter mixed with pungent and the texture was sticky solid. The yield of the extract was 9.1%. It was observed that crude extract contains carbohydrates, reducing sugars, monosaccharides, proteins, amino acids, fats and oil, steroids, cardiac glycosides, saponin glycosides, alkaloids, tannins and phenolic compounds, and flavonoids. The presence of many phytochemicals in the extract signifies its substantial contribution to various disorders. The crude extract was subjected to column fractionation using various solvents. For column fractionation, we used polar solvents and analyzed the resulting fractions using thin-layer chromatography (TLC) to determine the presence of individual or multiple compounds. From TLC analysis, it was observed that the ethanol fraction contains a single compound and therefore same fraction was subjected for HPLC, HPTLC, FTIR, and LC-MS analysis. From HPLC analysis, it was clearly identified that this ethanolic fraction contains chlorogenic acid and it displayed a peak at 8.428 minutes. The HPTLC analysis revealed the presence of chlorogenic acid. From HPTLC analysis, chlorogenic acid was identified at 0.65 Rf value. From the present investigation, we concluded that this developed method can be used for the isolation of chlorogenic acid from C. subcordata.