Total Flavonoid Content Research Articles

Study on the Mechanism of Flavonoid Enrichment in Black Soybean Sprouts by Abscisic Acid/Melatonin Under Slight Acid Treatment

Plant hormones play a critical role in the physiological and biochemical mechanisms of plants, with functions such as regulating the metabolic pathways of secondary metabolite production and alleviating external stresses. In this study, the synthesis of flavonoids in black soybean sprouts was induced by slight acid combined with the plant hormones abscisic acid (ABA) and melatonin (MT). The results indicated that the contents of daidzin, genistin, daidzein, and genistein in black soybean sprouts treated with slight acid were increased by 10 μM ABA and 75 μM MT, and the total flavonoid content was significantly enhanced. Compared with the slight acid treatment, the H2O2 and malondialdehyde (MDA) contents in black soybean sprouts were increased after ABA treatment, and the black soybean sprouts were further stressed. However, the H2O2 and MDA contents in black soybean sprouts were significantly decreased after MT treatment, indicating that the stress of black soybean sprouts can be alleviated by MT. Under slight acid stress, the genes related to flavonoid synthesis in black soybean sprouts were induced by exogenous ABA, promoting the accumulation of flavonoids; under exogenous MT treatment, the activity of phenylpropanoid metabolism enzymes was significantly increased, the genes related to flavonoid synthesis were upregulated, and flavonoid synthesis was induced. These results suggest that the combination of slight acid and plant hormone treatments promotes the accumulation of flavonoid substances during the germination of black soybeans. This research lays the foundation for improving the growth conditions of black soybeans and promoting the enrichment of flavonoid substances in black soybeans.

Assessment of Pudding Formulations Using Lyophilized Apricot, Plum, and Plum–Apricot Powders: Texture, Bioactivity, and Sensory Quality

The food sector is constantly responding to consumers’ increased demands concerning healthy nutrition and beneficial ingredients. This study presented the development of three pudding alterations using lyophilized fruit (apricots, plum–apricots, and plums) powders. The same concentrations of fruit powder fully substituted the sugar in each formulation. The results showed that each new formulation formed a thick gel consistency and had full water-holding capacity at 24 h of storage. The color differed according to the established CIE-lab data. The lightness varied from 42.57 ± 1.97 (pudding formulation using plum powder) to 81.91 ± 1.18 (control sample). The total soluble solids and titratable acidity showed that the control sample was different from each new formulation. The water activity was similar in all studied samples varying from 0.978 ± 0.003 to 0.989 ± 0.001, and the plum and plum–apricot formulations had a pH near the control samples, at 6.54 and 7.23, respectively. The antioxidant activity, total polyphenol content, and total flavonoid content were also evaluated. The ABTS assay revealed the highest results compared to the other three applied methods. The sensory evaluation showed that it is necessary to further improve the recipes for better consumer perception.

Influence of altitudes and development stages on the chemical composition and antioxidant capacity of Andean blackberries (Rubus glaucus Benth)

BackgroundThe Andean blackberry (Rubus glaucus Benth), locally known as “Mora de Castilla,” is a high-altitude fruit rich in phytochemicals, mainly anthocyanins, with potent antioxidant properties. Although its chemical composition has been studied, the influence of altitude on its phytochemical profile and antioxidant capacity is still unclear. This research aimed to investigate the effects of development stages and altitude on the phytochemical profile and antioxidant activity of this fruit.MethodsAndean blackberry fruits were collected at different altitudes and development stages in the central Andean region of Ecuador. An hydroalcoholic extraction was used to quantify the compounds and determine the total antioxidant activity, while solid phase extraction (SPE) was performed to separate anthocyanins from other compounds. Ultra-high-performance liquid chromatography coupled to a triple quadrupole mass spectrometer (HPLC-DAD/ESI-MSn) was used to identify anthocyanin and non-anthocyanin phenols, while total antioxidant capacity, total polyphenols, total flavonoid content, and total anthocyanin content were quantified spectrophotometrically.ResultsFlavonols and anthocyanins constitute the majority of the flavonoids identified in the Andean blackberry extracts, both in terms of peak areas and number of identified compounds, followed by ellagic and gallic acid derivatives, as well as phenolic acids, mainly hydroxycinnamic acids. Quercetin was identified as the predominant flavonol in unripe berries, where anthocyanins were not significantly present. On the contrary, in ripe berries, cyanidin and its glycosides stood out as the main anthocyanins and predominant compounds. We observed that in the early stages of ripening, the total polyphenol content predominates in the berries and is mainly responsible for their antioxidant capacity. However, as the fruit ripens, the total anthocyanin content increases, becoming the most prominent bioactive compounds in fully ripe berries.ConclusionThe results suggest that higher altitude environmental conditions may improve the composition, concentration of phenolic compounds, and antioxidant activity of Andean blackberries. Overall, our findings highlight the high functional value of this fruit, supporting its health-protective effects when consumed regularly, either as fresh fruit or in nutraceutical form.

Assessment of Bioactive Compounds and Physiological Activities of Ethanolic and Aqueous Extracts from Black Rice, Black Rice Bran, and Milled Black Rice

Black rice (BR) is considered one of the healthiest foods worldwide. This study assessed the bioactive compound contents and antioxidant activities of the distilled water (DW) and 80% ethanol (EtOH) extracts of BR, black rice bran (BRB), and milled black rice (MBR). In addition, their anti-obesity, anti-obesogenic, and anti-muscle atrophy effects were comparatively evaluated. The results revealed that the 80% EtOH extract of BR contained higher contents of β-glucan, total flavonoids, and total phenolics (26.97 ± 0.70 mg gallic acid equivalents (GAE)/g and 16.58 ± 0.37 mg quercetin equivalents (QE)/g, respectively) and showed higher antioxidant activity than those of the BRB and MBR extracts. On the other hand, BRB contained the highest content of γ-oryzanol, which was 15.12 ± 0.03 mg/g. Moreover, the 80% EtOH extract of BR exhibited a stronger anti-obesogenic effect by downregulating adipogenesis-related proteins (PPAR-γ, C/EBP-α, and FAS) and upregulating lipolysis-related proteins (PGC-1α and CPT-1), while the BRB extract effectively increased myotube diameter and fusion index. Overall, these findings indicate that the 80% EtOH extracts of BR, BRB, and MBR, particularly the BR extract, have high potential as natural antioxidants and anti-obesity agents, with BRB showing promise in preventing muscle atrophy.

Optimization of microwave-assisted extraction using response surface methodology and HPLC-DAD phenolic compounds quantification from Hylocereus undatus peel and pulp cultivated in Tunisia

The present study aimed to optimize the microwave-assisted extraction process for both the peel and pulp of Hylocereus undatus (white dragon fruit) cultivated in Tunisia, using response surface methodology. Total phenolic content, total flavonoid content, FRAP (ferric reducing antioxidant power), and DPPH (1,1-diphenyl-2-picrylhydrazyl) antioxidant activities were optimized. A central composite design (CCD) was applied, considering three key variables: extraction time, extraction temperature, and liquid-to-solid ratio. The optimized extraction parameters for Hylocereus undatus peel and pulp were determined respectively (9.57 min, 42.20 °C and 27.79 mL/g) and (10.08 min, 40.84 °C and 31.52 mL/g). The main phenolic compounds identified in Hylocereus undatus peel and pulp extracts using HPLC-DAD were chlorogenic and caffeic acids and rutin, quercetin, luteolin-7-O-glucoside as flavonoids. Therefore, this research has revealed the potential of a sustainable and eco-friendly process hold promise a directional option and encouraging a circular economy approach for industrial production of antioxidant-rich Hylocereus undatus extracts.

Bioactive Properties of Campomanesia lineatifolia: Correlation Between Anti-Helicobacter pylori Activity, Antioxidant Potential and Chemical Composition

Helicobacter pylori is found in the stomach of patients with chronic gastritis and peptic ulcers, infecting approximately half of the world’s population. Current treatment for H. pylori infection involves a multi-drug therapeutic regime with various adverse effects, which leads to treatment abandonment and contributes to the emergence of resistant strains of H. pylori. Previously, we demonstrated that the essential oil of Campomanesia lineatifolia leaves exhibited an anti-H. pylori activity. In this study, we aimed to evaluate the phenolic content of the phenolic-rich ethanol extract (PEE) from C. lineatifolia and its anti-H. pylori and antioxidant properties. Additionally, the anti-H. pylori activity was assessed in polar and non-polar fractions from PEE, isolated myricitrin (MYR) and a mixture of myricitrin and quercitrin (MYR/QUER) from polar fractions, and aqueous extract (tea) to correlate the responsible fractions or compounds with the observed activity. Broth microdilution assays were performed to assess the anti-H. pylori activity using type cultures (ATCC 49503, NCTC 11638, both clarithromycin-sensitive) and clinical isolate strains (SSR359, clarithromycin-sensitive, and SSR366, clarithromycin-resistant). The antioxidant activity was evaluated using the DPPH assay. The total tannin and flavonoid contents were determined using the hide-powder method, the Folin-Ciocalteu reagent, and the aluminium chloride colourimetric assay, respectively. The tea (MIC 1:100), PEE, polar and non-polar fractions, MYR, and MYR/QUER inhibited the growth of H. pylori strains tested (MIC values ranging from 0.49 to 250 μg/mL). The antioxidant assays revealed that PEE exhibited a higher antioxidant activity (EC50 = 18.47 μg/mL), which correlated to the high phenolic content (tannin and flavonoid, 22.31 and 0.15% w/w, respectively). These findings support the traditional use of C. lineatifolia as a multitarget medicinal plant for treating gastric ulcers and reinforce the potential use of the species as a coadjuvant in therapeutic regimes involving patients with resistant H. pylori infection.

Foliar Application of Methyl Jasmonate and Chitosan Improve Growth, Yield, and Quality of Guar (Cyamopsis tetragonoloba L.) Under Water-Deficit Stress

Guar (Cyamopsis tetragonoloba L.), a summer legume, is becoming increasingly important as an industrial crop due to its high gum and viscosity content. This study investigated the effects of methyl jasmonate (MeJA), chitosan (CH), and their combination on the growth, yield, and quality of guar under irrigation regimes. A greenhouse experiment was conducted using a factorial design to evaluate the effect of foliar spraying with MeJA (5, 25, and 50 µM), CH (100, 150, and 200 mg/L), their combination (25 µM MeJA + 150 mg/L CH), and control on two commercial guar varieties (RGC-986 and BR-2017) under different irrigation regimes (100%, 70%, and 40% field capacity). The results showed that the exogenous application of MeJA and CH, individually and in combination, significantly enhanced various morphological traits and yield components in guar, including plant height, pod characteristics, seed yield, and root development. Additionally, the combination treatments improved seed quality parameters, such as gum percentage and viscosity content. Leaf analysis revealed increased levels of total phenolic content, total flavonoid, and anthocyanin contents. The BR-2017 variety showed superior performance in most morphological and qualitative traits, demonstrating greater resistance to irrigation regimes. It maintained yield and quality characteristics under water-deficit conditions, particularly when treated with 25 µM MeJA and 150 mg/L CH. The highest gum percentage (33.67%) and viscosity (4768.5 cP) were observed in the RGC-986 variety, along with enhanced levels of secondary metabolites. This study provides new insights into how MeJA, CH, and their combination can improve the yield and quality of guar under water deficit stress conditions. The results suggest that the use of these elicitors, especially in combination, represents an innovative strategy for improving guar production and quality, with potential variety-specific responses to water-deficit stress.

Effect of vermicompost on the nutraceutical quality of tender cladodes of Opuntia ficus-indica

The consumption of tender cladodes knows as “nopalitos” (Opuntia ficus-indica L. Miller.) has increased worldwide. That is because it is an important nutritional source in the human diet given its content of fiber, minerals, vitamins, phenolic compounds and secondary metabolites (antioxidants, phenols and flavonoids), which are essential for human health. The objective of this study was to evaluate the effect of doses of vermicompost (0, 5, 15 and 25 t ha-1) on the yield and nutraceutical quality of tender cladodes of O. ficus-indica. The measured variables were tender cladodes production and nutraceutical quality. The results showed no significant effect on the content of total phenolic compounds, total flavonoids and antioxidant capacity. However, the application of vermicompost showed significant effects on the yield, in which higher estimates were achieved for the highest dose with up to 100% compared to the control. The data obtained indicate that the dose of 25 t ha-1 of vermicompost promotes and increase in yield without compromising the nutraceutical quality of “nopalitos”.

The Utilization of Vegetable Powders for Bread Enrichment—The Effect on the Content of Selected Minerals, Total Phenolic and Flavonoid Content, and the Coverage of Daily Requirements in the Human Diet

The aim of this study was to evaluate the content of selected minerals and total phenolic and flavonoid content of wheat bread and bread enriched with varying amounts of carrot powder (CP) and pumpkin powder (PP). In addition, the coverage of daily requirements of selected minerals was evaluated after the consumption of 100 g of each type of bread. The research included seven types of bread: wheat bread (WB) and bread enriched with 10%, 20%, and 30% CP and PP. The vegetable powders were obtained by freeze-drying. The concentrations of minerals—sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), copper (Cu), and manganese (Mn)—were determined by atomic absorption spectroscopy (AAS) method. Coverage of daily requirements after consumption of a serving of bread was determined based on Recommended Daily Allowance (RDA) and Adequate Intake (AI) ratios for minerals, according to the nutritional standards for the Polish population. The addition of 10% PP led to a greater increase in Na, K, Mg, and Cu in the bread compared to CP, while for Ca, Fe, Zn, and Mn a greater increase was observed with the addition of 10% CP than PP. Among macronutrients, the greatest changes in content were recorded for Ca—the addition of 10 and 20% CP and PP resulted in increases of 66, 113, 51 and 59%. Among micronutrients, the addition of CP and PP to wheat bread caused the largest changes in Cu (46–150% increase) and Mn (25–99% increase) content. Additionally, there was a tenfold increase in total phenolic content (TPC) when 30% CP was added to the bread. Consuming 100 g of bread with CP and PP provided the greatest coverage of K (41–60%), Cu (8–17.5%), and Mn (6–17%). These findings suggest that fortification of bread with vegetable powders has the potential to be a useful method of enriching the mineral composition and increasing the proportion of selected minerals in the human diet.

Qualitative and quantitative analysis of major components in Abelmoschus manihot flowers treated with different drying methods using UHPLC Q-Exactive MS and HPLC-PDA

The flowers of Abelmoschus manihot (L.) Medic are commonly used in clinical practice in China to cure forms of chronic kidney disease. Despite a long history of traditional use, the flowers obtained by different drying technologies have never been fully chemically characterized, and the ranges of constituents between different drying methods have not been comprehensively reported. To establish a quality control and chemical characterization method, a total of 14 batches of samples corresponding to 14 postharvest treatments were studied. Seven flavonoids were quantified using a HPLC-PDA method. The method was validated in terms of linearity (r > 0.999), precision (intra- and inter-day: 0.7–1.4%), accuracy (99.90–100.7%), detection limit (0.34–0.46µg/mL) and quantification limit (1.15–1.52µg/mL). The contents of total flavonoids in manihot flowers were as follows in descending order: Infrared Drying (50.96mg/g) > Microwave Drying (41.84mg/g) ≈ Hot-air Drying (39.58mg/g) ≈ Fresh (39.35mg/g) ≈ Primary Drying (38.95mg/g). Principal component analysis showed that samples processed with Fresh, Primary Drying, and the investigated three modern drying methods were well classified into three domains, indicating an important difference between drying methods. For the purpose of saving the flavonoids contents, infrared drying under 80~100 ℃ would be most acceptable. Furthermore, using UHPLC Q-Exactive Orbitrap MS data with targeted and non-targeted approaches, 28 compounds were identified in Abelmoschus manihot samples. Flavonoids were the main group of compounds found in Abelmoschus manihot flowers. The study could provide the scientific evidence for the selection and optimization of appropriate drying method for manihot flowers, and also provide the reference for the formation of generic primary drying processing technology for medicinal flowers containing flavonoids.

Change Patterns of Major Functional Components in the Loquat Summer Shoot Leaf during Annual Growth

Loquat leaves are widely used in traditional Chinese medicine and food processing as raw materials. To provide a theoretical basis for suitable harvesting periods based on the utilization of distinct components, the change patterns of major functional components in the annual growth of loquat leaves were investigated. In this study, the summer loquat leaves were sampled regularly during their annual growth, and the contents of seven main functional components and 13 monomer components were determined. The results showed that there was a regular change pattern for major functional components during loquat annual growth from Jul 2020 to Jun 2021; however, the change trend was different among different components. Among them, the contents of total flavonoids, total phenols, total triterpenoid acids, vitamin C, asiatic acid, quillaic acid, corosolic acid, and chlorogenic acid presented an overall upward trend. The content of maslinic acid showed a downward trend, whereas the content of neochlorogenic acid did not change much throughout the whole growth period. The content changes in the loquat leaves were affected by the light durations and temperature changes in different seasons, with mutual transit during the flowering and fruit growth processes. Sampling in May to June was found to be a most optimized option when applied in Chinese medicine processing; however, the degree of fibrosis is another indicator when it is used as food raw material. This study revealed the change pattern of major functional components of loquat leaves during the annual growth period and provides a theoretical basis for the reasonable harvesting period of loquat leaves as raw materials for processing different types of products.

Evaluation of the antioxidant activity and prebiotic properties of mangosteen peel proanthocyanidin extracts through simulated in vitro digestion and colonic fermentation

The biological activity of mangosteen peel proanthocyanidin extracts is related to digestive properties and colonic metabolism. Therefore, the study's aim was to investigate the dynamics of proanthocyanidin extracts during in vitro digestion and colonic fermentation, their protective effects against oxidative damage in PEC-J2 cells, and their interactions with the intestinal flora. The results showed that epicatechin and proanthocyanidin B2 were the most abundant in proanthocyanidin. In addition, the catechin was increased by 27.7 ± 0.2 μg/mL. The total phenol content, total flavonoid content, and proanthocyanidin content of mangosteen peel were reduced by 32.36%, 90.63%, and 78.06%, respectively. However, the antioxidant capacity was increased by two-fold, protecting against oxidative damage in cells. Proanthocyanidin polymers that are not digested by the gastrointestinal tract go to the colon, where they are metabolized by microorganisms to produce short-chain fatty acids (SCFAs). Proanthocyanidin extracts modulated the microbial flora structure: the relative abundance of Phascolarctobacterium, Roseburia, Faecalibacterium, Coprococcu, and Clostridium sensu stricto all increased significantly. The ratio of Bacteroides to Firmicutes increased, and Bifidobacterium grew and multiplied. This study improves the utilization of mangosteen peel and provides a theoretical basis for the scientific evaluation of the antioxidant activity and gut microbiota modulation of mangosteen peel.

Eco-Friendly Synthesis of Silver Nanoparticles: Enhancing Optimization Reaction, Characterization, and Antimicrobial Properties with Lantana camara from Geothermal Area

Green synthesis methods for producing silver nanoparticles (AgNPs) have garnered significant attention for their potential in medical applications. Despite the known potential of Lantana camara in AgNP synthesis, there is a lack of studies investigating its application when grown in extreme geothermal environments, which may influence the properties and efficacy of the synthesized nanoparticles. This research aimed to fabricate AgNPs utilizing aqueous extract from Lantana camara, a plant growing in an extreme geothermal manifestation area. Another aim of this study is to evaluate their antimicrobial activity. Qualitative and quantitative phytochemical analyses of the plant's leaves were also conducted. Reaction optimization was performed using response surface methodology (RSM), employing a central composite design (CCD) approach. The characterization of AgNPs involved UV-Vis spectroscopy, FTIR, SEM-EDX, and PSA. The antimicrobial testing was conducted against Gram-positive bacterium (S. aureus), Gram-negative bacterium (E. coli), and the fungus (C. albicans). The phytochemicals analysis revealed that the L. camara leaf extract contains flavonoids, phenolics, saponins, tannins, and steroids, lacking alkaloids and terpenoids, with total phenolic and flavonoid contents of 11.94 mg (GAE/g) and 6.70 mg (QE/g), respectively. The AgNPs exhibited a spherical shape with a surface plasmon resonance (SPR) peak at a wavelength of 417 nm, and the smallest particle size measured was 44 nm. Based on FTIR analysis, AgNPs have functional groups such as OH, NH, C=C, and CH that were identified as groups involved in the reduction of Ag+ to Ag0 during the green synthesis of AgNPs. The AgNPs demonstrated the lowest antifungal activity against C. albicans. In summary, the aqueous leaf extract of L. camara from geothermal manifestation areas can serve as a bioreductant for AgNPs, exhibiting higher antibacterial activity against Gram-positive bacteria compared to Gram-negative ones.

Anti-inflammatory, antioxidant, and anti-apoptotic properties of Thunbergia laurifolia Lindl. aqueous leaf extract on LPS-stimulated RAW 264.7 macrophages

Thunbergia laurifolia Lindl., a Thai medicinal herb, has garnered substantial attention among researchers due to its recognized efficacy as an antidote, antioxidant, and anti-inflammatory agent. Despite its extensive investigation, the specific role of T. laurifolia in apoptotic inhibition remains unexplored. Therefore, this study aimed to explore the protective properties of T. laurifolia leaf extract against lipopolysaccharide (LPS)-induced apoptosis in RAW 264.7 macrophages and determine the extractʼs total phenolic as well as flavonoid content, and its antioxidant. Moreover, the cytotoxicity including anti-inflammatory potential on LPS-stimulated RAW 264.7 macrophages were assessed by measuring the inhibition of nitric oxide (NO). The outcomes of this investigation revealed remarkable similarity between the total phenolic and flavonoid contents and antioxidant capacity of the leaf extract under scrutiny and those documented in prior studies. In addition, the extract demonstrated negligible cytotoxicity at low concentrations. Furthermore, it effectively impeded the LPS-mediated production of NO, inflammatory mediator, in the macrophages. Moreover, pre-treatment with T. laurifolia leaf extract improved LPS-induced apoptosis by reducing rate of apoptotic cell death up to 41 % at the same concentration (1.68 μL/mL) in LPS-stimulated RAW 264.7 macrophages. Collectively, these findings provide compelling initial evidence attesting to the ability of T. laurifolia leaf extract to inhibit LPS-induced apoptosis in RAW 264.7 macrophages, thereby augmenting our understanding of its potential therapeutic applications in inflammatory conditions.

Effect of Biostimulants on Plant Growth and Leaf Functional Compounds of Passiflora Plants

The influence of biostimulant treatments on Passiflora species was assessed by observing changes in the growth and leaf functional compounds of plants exposed to humic acids (HAs) and seaweed extracts (SEs) to determine their optimal application doses. Four different concentrations of HA irrigation (0, 144, 192, and 285 mg·L−1 per pot assigned as HI-1, 2, 3, and 4, respectively), HA spraying (0, 96, 128, and 192 mg·L−1 as HS-1, 2, 3, and 4, respectively), SE irrigation (SI-1∼4 at 0, 1, 1.33, and 2 g·L−1 per pot, respectively), and SE spraying (SS-1∼4 at 0, 0.5, 0.67, and 1 g·L−1, respectively) treatments were applied to ‘Tainung No. 1’ and Passiflora suberosa cultivars every week to study their responses in plant growth traits and metabolites. Results show that leaf dry weight (DW) and root DW of ‘Tainung No. 1’ plants under SS-3 and HI-4 treatments, respectively, were significantly increased compared with controls. Both HI-3 and HS-3 treatments remarkably increased vine length, fresh weight (FW), and DW of leaves and shoots in P. suberosa plants compared with controls. Furthermore, ‘Tainung No. 1’ and P. suberosa plants respectively subjected to HI-2 and SS-4 treatments contributed to developing biostimulant applications for obtaining higher total phenol and flavonoid content per plant. Both Passiflora species are recommended for treatment with SS-4 and SE to maximize orientin and isovitexin content, and for all their beneficial applications and sustainable use in agriculture.

Determination of Total Flavonoid Content from Extract and Fractions of Mangrove Leaves (Rhizopora mucronata)

Indonesia is an archipelagic country with a very wide coastal area. Lombok Island has a coastal area with quite extensive mangrove forest areas. Rhizopora mucronata was the most dominant species found in the region. The Rhizopora mucronata species was a plant from the Rhizoporaceae family reported to have antibacterial, antifungal, and antioxidant activity. These biological activities were caused by the presence of secondary metabolite compounds in R. mucronata, one of which is flavonoids. Research regarding the flavonoid content of R. mucronata leaves has not been explored. Therefore, the study aimed to determine the total flavonoid content in extracts and fractions of R. mucronata leaves using the spectrophotometric UV-Vis method. R. mucronata mangrove leaves simplicia were extracted using the sonication method using 96% ethanol solvent, then fractionated using the liquid-liquid extraction method with n-hexane, ethyl acetate, and water. The percentage yield of 96% ethanol extract, water fraction, ethyl acetate fraction, and n-hexane fraction respectively were 42.33%; 43.147%; 40.49%; 9.74%; and 32.49%. The TLC test showed the presence of flavonoid compounds detected in the extract and fractions which were marked with blue spots. The total flavonoid contents in the 96% ethanol extract, water fraction, ethyl acetate fraction, and n-hexane fraction were respectively (12,980; 14,160; 23,880; and 25,350) mg Quercetin equivalent/gram sample (mg QE/ g). The total flavonoid content of the extract and fractions of R. mucronata leaves were analyzed using the One-way ANOVA test method. The total flavonoid content in the n-hexane fraction and ethyl acetate fraction was significantly higher than the 96% ethanol extract and water fraction. The total flavonoid content of R. mucronata leaves was relatively high, which contributes to several of its biological activities.

Unveiling the influences of P fertilization on bioactive compounds and antioxidant activity in grains of four sorghum cultivars.

Phosphorus is a critical nutrient in agriculture, influencing plant growth and nutritional quality. This study, uniquely designed to investigate the effects of phosphorus (P) fertilization levels, sorghum cultivars, and growing locations on phytochemical content and antioxidant activity in sorghum grains, employed four sorghum cultivars (Hakeka, P954063, Tabat, and Tetron) grown under three P levels (0P, 1P, 2P) in two locations (Gezira and White Nile) in Sudan. In this study, four sorghum cultivars were grown in two distinct locations in Sudan, employing a split-plot design with three (P) fertilization levels. P was applied as triple super phosphate directly with the seeds, and additional fertilization included urea applied in two split doses. At physiological maturity, representative sorghum panicles were harvested, processed, and analyzed for bioactive compounds and antioxidant activities using standard extraction and quantification techniques such as Folin-Ciocalteu for phenolics and colorimetric flavonoid assays. Antioxidant activities were assessed through various assays, including DPPH and FRAP. Statistical analyses were performed using a three-way ANOVA to examine the effects of cultivar, P level, and location on the measured parameters, supplemented by multivariate analysis to further elucidate the interactions between these factors. Significant interactions (p<0.001) were observed among cultivars, P levels, and locations for total phenolic content (TPC), total flavonoid content (TFC), carotenoids, tannins, and various antioxidant activity measures (DPPH, FRAP, ABTS, TRP, H2O2). P fertilization significantly increased all measured phytochemicals and antioxidant activities compared to non-treated cultivars, except for H2O2, which decreased with P application. Among cultivars, Hakeka consistently exhibited the highest TFC, carotenoid content, and antioxidant activities (DPPH, FRAP, TRP, ABTS), particularly at the 2P level. P954063 showed the highest TPC and tannin concentrations. Tetron generally had the lowest phytochemical and antioxidant levels. White Nile showed higher TPC, carotenoids, DPPH, FRAP, TRP, and ABTS levels, while Gezira had higher TFC, tannins, and H2O2 concentrations. The impact of phosphorus fertilization often varies between locations. Strong positive correlations were found between TPC and all antioxidant assays (r = 0.68-0.90) and total carotenoids and antioxidant activities (r = 0.73-0.93). This study recommended cultivating the Tabat variety with 2P doses in Gezira. In addition, the Hakeka cultivar showed the highest increases in total flavonoid content, carotenoids, and antioxidant activities, particularly under the highest P level (2P). The findings highlight that P plays a critical role in enhancing sorghum's nutritional and health-promoting qualities, which are essential for leveraging this staple crop for food and nutrition security strategies in semi-arid regions.

Evaluation of Antioxidant and Breast Cancer Cytotoxicity of Beta vulgaris and Daucus carota Methanol Extracts

Breast cancer is the world's most frequent malignancy among women. Many dietary antioxidants have been shown to help prevent oxidative stress, which has been linked to a variety of diseases, including cancer. Many chronic diseases, such as cancer, can be avoided by eating fruits and vegetables on a regular and balanced basis. The current study's goal is to assess the antioxidant and cytotoxic capabilities of methanolic extracts of Beta vulgaris and Daucus carota against MCF-7 human breast cancer cell lines. Therefore, the aim of the present study was to determine the phytochemical components, antioxidant capacity, total phenolic &amp; flavonoid contents in the methanolic extracts of Beta vulgaris and Daucus carota. Further, MTT assay against breast cancer cell line (MCF-7) was performed for the evaluation of cytotoxic activity. The majority of secondary metabolites were found in both the aqueous and methanolic extracts of Beta vulgaris and Daucus carota. Beta vulgaris and Daucus carota have different total phenolic and flavonoid contents. TLC results showed many spots having different Rf values. Moreover, we observe comparable cytotoxic activity in both the extracts against MCF-7 cell lines. Our results reveal that the methanolic extracts of Beta vulgaris and Daucus carota has effective phytochemical constituents, antioxidant and anticancer activity. The total phenolic and flavonoid contents vary between Beta vulgaris and Daucus carota. Further studies are needed to evaluate the chemopreventive potentials of the Beta vulgaris and Daucus carota extract when used alone or in combination with doxorubicin to mitigate the toxic side-effects of the latter.

Investigating Flavonoids by HPTLC Analysis Using Aluminium Chloride as Derivatization Reagent

This is the first study to report on high performance thin layer chromatography (HPTLC) generated spectrophotometric data to systematically capture flavonoid compounds using optimized derivatization with either AlCl3 or NaNO2-AlCl3-NaOH as visualisation reagents. While the traditional AlCl3 colorimetric method using UV–Vis analysis provides valuable insights into the presence of flavonoids and allows derivation of the total flavonoid content (TFC) of a sample, HPTLC fingerprints obtained after spraying with AlCl3 or NaNO2-AlCl3-NaOH enable the visualization of the various flavonoids present in a sample based on their respective absorption shifts, thus complementing the traditional TFC assay. In this study, 40 different flavonoids representing different classes (flavonols, flavanolols, flavan-3-ol, flavones, flavanones, and isoflavonoids) were analysed. Upon derivatization with AlCl3 most of the investigated flavonoids recorded bathochromic shifts, yielding characteristic λmax values between 370 and 420 nm, while spraying with NaNO2-AlCl3-NaOH triggered hyperchromic shifts, and thus an increase in absorbance intensity in flavonoids with particular substitution patterns. A few non-flavonoid components with structural similarities to flavonoids (e.g., rosmarinic acid, gallic acid, aspirin, salicylic acid) served as the negative control in this study to determine whether the derivatization reagents allowed exclusive detection of flavonoids. The method was then applied to the analysis of flavonoid containing supplements as well as red clover honey to demonstrate the method’s application in the analysis of natural products.

Integrated Metabolome and Transcriptome Analysis Reveals the Key Bitter Substances of Pleioblastus amarus Shoots at Different Developmental Stages

Pleioblastus amarus (bitter bamboo) shoots are a traditional forest vegetable in China, renowned for its nutritional composition and associated health benefits. This study aimed to investigate the dynamic changes in nutritional quality and bitterness-related metabolites, as well as corresponding gene expression in P. amarus shoots at three distinct growth stages, specifically at heights of 10 cm (BT1), 20 cm (BT2), and 40 cm (BT3). The results showed that the content of soluble sugars decreased while the content of total phenols and flavonoids, which contribute to bitterness, increased during the growth of P. amarus shoots. In addition, comparative analysis of BT2 vs. BT1, BT3 vs. BT1, and BT3 vs. BT2 revealed 14, 43, and 36, respectively, differentially accumulated metabolites (DAMs). Notably, BT2 exhibited upregulation of flavonoids, such as apigenin, while bitter amino acids, including phenylalanine and arginine, showed a marked increase in BT3. Moreover, differential gene expression analysis revealed 3161, 13268, and 3742 differentially expressed genes (DEGs) in the BT2 vs. BT1, BT3 vs. BT1, and BT3 vs. BT2 comparisons, respectively. An integrated analysis of metabolomic and transcriptomic data indicated that the metabolites and genes associated with the biosynthesis pathways of phenylalanine, tyrosine, tryptophan, arginine, and the branched-chain amino acids valine, leucine, and isoleucine were significantly enriched during the growth of P. amarus shoots. The study indicates that early-stage (≤20 cm) P. amarus shoots are mainly bitter due to flavonoids like apigenin, while the bitterness in later-stage (around 40 cm) shoots is largely due to bitter amino acids such as phenylalanine, arginine, valine, and isoleucine. This research highlights key compounds and genes related to bitterness, providing a basis for future research on enhancing the nutritional value and flavor of P. amarus shoots.