Screening Factorial Design for Extraction of Bioactive Metabolites from Tithonia diversifolia (Hmsl.) A. Gray/Asteraceae Leaf Extract: Phytochemical Assays and Chemical Characterization by Synchronous Fluorescence and Phosphorescence Spectroscopy
Novel SFS, SPS and 3D-SFS are described as fingerprinting/comparative profiling techniques for leaf extracts which require definitive compound identification methods.
Abstract
Recent studies highlight the rich medicinal flora of São Tomé and Príncipe (STP) islands due to their therapeutic potential against various diseases. To maximize bioactive compounds’ extraction from Tithonia diversifolia leaves, we have used a two-level, three-factor (23) full factorial design. The screening evaluated water and hexane solvents at 25 °C and 40 °C, across incubation periods of 0 and 5 days, maintaining a constant agitation of 150 rpm. The highest TPC among the tested conditions was 72.16 µmole gallic acid equivalent/g of leaves using water at 40 °C with a 5-day incubation period. The lea extracts were characterized through several phytochemical assays, including TFC, reducing power, ABTS, DPPH, and SOD activity. This report introduces a novel, non-destructive analytical fingerprinting approach using intrinsic synchronous fluorescence and phosphorescence. Intrinsic, non-destructive synchronous fluorescence spectroscopy was performed across a wavelength range of 250 to 750 nm, utilizing a Δ λ interval of 5 to 30 nm. The hexane leaf extract displayed distinct peaks at 290, 320, 345, 400, 490, and 675 nm. Conversely, the aqueous extracts only showed peaks at 490, 560, and 675 nm. Intrinsic synchronous phosphorescence testing revealed peaks at 325, 400, 490, 550, and 675 nm for the hexane extract, and at 500 and 560 nm for the aqueous extract. Additionally, 3D fluorescence fingerprinting spectra successfully validated the emission peaks at 290, 320, 345, 400, 490, and 675 nm. Therefore, the present work describes novel SFS, SPS and 3D-SFS as fingerprinting/comparative profiling techniques for leaf extracts which require definitive compound identification methods.
ABSTRACT Alternanthera bettzickiana exhibits considerable morphological diversity, which has contributed to significant inconsistencies in reported phytochemical profiles and bioactivities, as the majority of published studies have not specified the morphotype of the plant material examined. Unlike the green morphotype, which is widely utilized in culinary applications, the variegated (mixed red and green) morphotype has received considerably less attention as an edible vegetable. Furthermore, no validated extraction protocol has been established for this species, despite being a critical determinant of bioactive compound recovery. The present study therefore aimed to optimize extraction conditions and compare the phenolic profiles and antioxidant capacities of both morphotypes. Utilizing Box–Behnken design (BBD)‐based response surface methodology (RSM), the optimized extraction conditions comprised 90% (v/v) aqueous ethanol, a shaking temperature of 50°C, an incubating time of 2 h, and an extract concentration of 10 mg/mL. Under these conditions, the green morphotype exhibited higher total phenolic content (4.95 mg gallic acid equivalent/g dry sample weight (DW)), whereas the variegated morphotype exhibited higher total flavonoid content (31.95 mg quercetin equivalent/g DW) and total anthocyanin content (7.04 mg cyanidin‐3‐O‐glucoside equivalent/g DW). Liquid chromatography‐electrospray ionization‐tandem mass spectrometry identified p‐coumaric acid as the predominant phenolic compound, followed by 3,4‐dihydroxybenzoic acid, in both morphotypes, while rutin was exclusively detected in trace amounts in the green morphotype. The green morphotype also demonstrated significantly higher ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) activities, whereas 2,2‐diphenyl−1‐picrylhydrazyl (DPPH) radical scavenging activities were not significantly different between morphotypes. These findings establish a scientific basis for the future valorization of both morphotypes as functional food ingredients and nutraceutical sources.
This study evaluates the thin-layer desorption kinetics, isolation-stabilization parameters, and bioactive efficacy of secondary metabolites from Parameria laevigata leaf matrices, mapping its chemical potential as a functional hepatoprotective beverage. Fresh biomass was processed using forced-convection thermal drying at 125°F for 4 hours and 140°F for 3 hours, with both regimes reaching a stable moisture desorption equilibrium at a 74% mass reduction (5% residual moisture). Phytochemical profiling and multi-laboratory screening confirmed that the low-temperature 125°F parameters protected fragile organic compounds from thermal oxidation, safely preserving high concentrations of total flavonoids, saponins, and condensed tannins along with essential micronutrients. Blind sensory screening by 19 independent evaluators confirmed that this gentle drying profile optimized the preservation of natural volatile compounds, yielding a darker color liquor, enhanced solute extraction, and superior aromatic qualities. Preclinical in vivo assays using carbon tetrachloride (CCl4)-induced hepatotoxicity models in male Sprague Dawley rats confirmed that a 100% concentration of the natural product extract completely blunted pathological serum enzyme elevations (SGOT and SGPT), maintaining baseline levels within normal parameters (p < 0.05). Microscopic histopathological reviews corroborated this systemic protection, showing that the isolated polyphenolic and metabolite matrix effectively prevented lobular disarray, severe necrosis, and vascular space dilatation, exhibiting structural tissue regeneration equal to the standard clinical reference drug Silymarin (100 mg/kg). These findings validate the natural product chemistry profile of P. laevigata as a stable, standardized, and highly viable bio-accessible formulation for targeted hepatoprotection.
Jandy S. Danzalan, Generaldo D. Maylem, Julius T. Capili· Genetics and Molecular Resea...· 0 citations
Objectives: Tinospora cordifolia, a medicinal plant, was widely used as traditional therapeutics, but the phytochemicals and bioactivities remained underexplored in a comparative fresh versus dry stem context using advanced analytical approaches. Using a combination of analytical and biological methods, T. cordifolia’s phytochemical content and bioactive potential were examined. Standard phytochemical screening and spectroscopic, chromatographic, and biological assay methods were used in this study.
Methods: The total phenolic and flavonoid content of T. cordifolia fresh and dry stems was quantitatively estimated. Fourier-transformed infrared (FTIR) spectroscopy was used for functional group analysis, while gas chromatography and mass spectroscopy (GC-MS) analysis were employed for chemical constituent identification, and the antioxidant property of T. cordifolia stem was analyzed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and phosphomolybdenum tests. The antimicrobial assay was assessed in relation to particular strains of bacteria and fungi.
Results: Phytoconstituents such as alkaloids, tannins, flavonoids, phenolics, saponins, and glycosides were found by quantitative analysis. While GC-MS profiling revealed a complex composition of fatty acid esters, phytosterols, terpenoids, and alkaloids, FTIR analysis confirmed amine, hydroxyl, and carbonyl moieties’ presence. Particularly in polar extracts, the DPPH and phosphomolybdenum assays and antimicrobial study demonstrated strong concentration-dependent action, superior radical scavenging capacity, and broad-spectrum antimicrobial potential, respectively.
Conclusion: The results support the traditional medicinal usage of T. cordifolia because of its high bioactivity, antibacterial properties, and abundance of natural antioxidants. The research offers a comprehensive phytochemical and bioactivity profile of T. cordifolia, integrating spectroscopic, chromatographic, and biological analyses. It highlights the plant’s potential for pharmaceutical and nutraceutical applications and lays the foundation for future studies on compound isolation, mechanism elucidation, and in vivo validation.
SHARMILA KJ, SANTHIYA JAYAKUMAR, NABEEL MOHAMMED· Asian Journal of Pharmaceuti...· 0 citations
