Optimization of Ultrasound-Assisted Extraction of Verbascum sinaiticum Leaves: Maximal Phenolic Yield and Antioxidant Capacity.

Verbascum sinaiticum (Qetetina or yeahya Joro) is a medicinal plant with secondary metabolites such as phenolics, flavonoids, glycosides, saponins, and alkaloids. This study was designed to optimize the ultrasonic-assisted extraction (UAE) parameters to enhance the phenolic content and characterize the phenolic compounds using ultra-high-performance liquid chromatography, coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC-ESI-QTOF-MS/MS), and antioxidant activities in Verbascum sinaiticum extract. Extraction time, sample-to-solvent ratio, and extraction temperature were considered for UAE optimization. It was found that UAE generated the highest extraction yield (21.6%), total phenolic content (179.8 GAE mg/g), total flavonoid content (64.49 CE mg/g), DPPH (61.85 µg/mL), and ABTS (38.89 µg/mL) when compared to maceration extraction. Metabolite analysis in this study showed the detection of 17 phenolic compounds, confirming antioxidant capacities. The optimization parameters have significant effects on phenolic compounds. Scanning electron microscopy showed the presence of structural changes when UAE was used over the maceration method. The optimized UAE parameters for extraction temperature (41.43 °C), sample-to-solvent ratio (36.32 g/mL), and extraction time (33.22 min) for TPC were obtained. This study shows the potential application for UAE of Verbascum sinaiticum leaves in the development of pharmaceutical and nutraceutical products.

Evaluation of Wound Healing Activity of Hydromethanolic Crude Extract and Solvent Fractions of the Root of Verbascum Sinaiticum Benth. (Scrophulariaceae) in Swiss Albino Mice.

Background: The roots of Verbascum sinaiticum have been used traditionally for the management of wound in different regions of Ethiopia. Despite the presence of several claims and in vitro studies regarding its role in wound healing, no scientific studies have been conducted so far. Therefore, this study aims to scientifically evaluate the wound healing activities of the crude extract and solvent fractions of the roots of Verbascum sinaiticum in Swiss albino mice. Methods: The dried root powder of Verbascum sinaiticum was extracted using 80% methanol by maceration technique. This was then fractionated with chloroform, ethyl acetate, and water. These extracts were formulated as ointment at 5% and 10% concentration by using simple base. Acute dermal toxicity was performed on mice. The wound healing potential was evaluated using excision, incision, and burn wound models. Results: In excision wound, 10% and 5% of crude extract ointment provided a significant (P<0.001) percentage of contraction starting from day 4 and day 6 onwards respectively. Moreover, the rate of epithelialization was significantly (P<0.001) improved in 10% crude extract. In burn wound, 10% and 5% crude extract showed significant (P<0.001) wound contraction starting from day 4 and 8 onwards respectively. In both excision and burn wounds, a moderate concentration of fibroblast proliferation and collagen deposition was observed on the 10% crude extract. The 5% and 10% aqueous and ethyl acetate fractions produced a significant (P<0.001) percentage of wound contraction and shortening of epithelialization at different time points compared to simple ointment. Conclusion: The results of this study demonstrated that the 80% methanolic crude extract, aqueous and ethyl acetate fractions of Verbascum sinaiticum root have wound healing potential which assimilates its traditional use.

In vitro antimicrobial and antioxidant activities, essential oil composition, and in silico molecular modeling analysis of secondary metabolites from roots of Verbascum sinaiticum.

Verbascum sinaiticum is locally used to treat wound, stomachache, viral infection, cancer, sunstroke fever, abdominal colic, diarrhea, hemorrhage, anthrax, and hepatitis. The objective of this study was to identify the compounds and to evaluate the antimicrobial and antioxidant activity of the extracts and isolated compounds from V. sinaiticum. The 1H-NMR, 13C-NMR, and DEPT-135 were used to elucidate the structures of isolated compounds. Essential oils were extracted by hydrodistillation method and their chemical analyses were performed by GC-MS. The broth microdilution method was used to evaluate the antimicrobial activity. The radical scavenging activity of the extracts and isolated compounds were evaluated using DPPH method. Silica gel column chromatographic separation of root extracts afforded seven known compounds: 3'-(4''-methoxy phenyl)-3'-oxo-propionyl hexadecanoate (1), harpagoside (2), pulverulentoside I (3), scrophuloside B4 (4), scropolioside A (5), scropolioside-D2 (6), and harpagide 6-O-ß-glucoside (7), which are all reported from this species for the first time. The EO extracts from leaves and roots were the most susceptible to Streptococcus agalactiae, with a 2 mg/mL MIC. The EO from roots was effective against Candida albicans and Trichophyton mentagrophytes, with a MIC of 8 mg/mL. The MeOH and CH2Cl2/CH3OH (1:1) root extracts showed the maximum activity against S. epidermidis with MIC values of 0.25 mg/mL. The strongest antibacterial effects were demonstrated against Staphylococcus epidermidis, which exhibited a 0.0625 mg/mL MIC for compound 1. The strongest radical scavenging activity was exhibited by the methanol extract (IC50 = 3.4 µg/mL), and compounds 4, 6, 5, 3, 7, and 2 with IC50 values of 3.2, 3.38, 3.6, 3.8, 4.2, and 4.7 µg/mL, respectively, in comparison with ascorbic acid (IC50 = 1.3 µg/mL). The results of the molecular docking analysis of compounds revealed minimal binding energies range from -38.5 to -43.1 kJ/mol, -33.1 to -42.7 kJ/mol, -34.7 to -39.3.7 kJ/mol, -25.5 to -37.6 kJ/mol against human myeloperoxidase (PDB ID: 1DNU), murA enzyme (PDB ID: 1UAE), human topoisomerase IIß (PDB ID: 4fm9), S. epidermidis FtsZ (PDB number: 4M8I) proteins, respectively. The docking results and the in vitro antibacterial activity are in good agreement. These findings show that the isolated compounds 2-7 can act as potential antioxidants and strong antibacterials against Staphylococcus aureus and S. epidermidis. As a result, V. sinaiticum root extracts have the potential to be effective in treating diseases caused by bacteria and free radicals, as long as further investigation has been suggested for the ultimate decision of this plant's potential candidate.

Antibacterial and antioxidant potential analysis of Verbascum sinaiticum leaf extract and its synthesized silver nanoparticles.

The potential applications of plant extract and nanoparticles in antibacterial and antioxidant studies have garnered significant interest in recent times. Despite being utilized in Ethiopian traditional medicine, Verbascum sinaiticum (qetetina) constituents and its usage in nanoparticle synthesis remain relatively unexplored. This study explores the potential of the plant extract and its nanoparticles for antibacterial and antioxidant applications, with a focus on the leaf extracts and its silver nanoparticles. The leaf extract was analyzed using LC-MS and GC-MS and found to contain over 70 compounds, including glycosides, phenolic compounds, flavonoids, and fatty acids. The synthesized nanoparticles had a maximum absorbance of 408 nm, with a size range of 2-40 nm and showed a spherical shape. Using the agar well diffusion method, the extract and nanoparticles were evaluated against Gram-positive (Staphylococcus aureus ATCC 2592, Streptococcus agalactia ATCC12386) and Gram-negative bacteria (Acinetobacter baumannii ATCC19606, Pseudomonas aeruginosa ATCC27853) bacterial strains. In terms of antibacterial effects, both the silver nanoparticles and leaf extract displayed a greater impact on gram-positive bacterial strains over gram-negative bacterial strains. Additionally, the tests for lowest inhibitory and bactericidal concentration indicated similar outcomes. Notably, the silver nanoparticles exhibited greater antibacterial activity compared to the leaf extract alone. The DPPH (2, 2-diphenylpicrylhydrazyl) assay was conducted to investigate antioxidant activity. The results showed that the plant extract had an IC50 value of 143 µg/ml, while the synthesized nanoparticle had an IC50 value of 216 µg/ml, indicating that the plant extract had greater antioxidant activity than the synthesized silver nanoparticles.

Antidiabetic Activities of 80% Methanol Extract and Solvent Fractions of Verbascum Sinaiticum Benth (Scrophulariaceae) Leaves in Mice.

Background: Because of the scarcity, high cost, and severe side effects of current medications, it is necessary to discover novel, safe, and affordable anti-diabetic drugs. The current study was conducted to evaluate the antidiabetic activities of Verbascum sinaiticum Benth leaves in mice. Methods: Leaf coarse powder was extracted with 80% methanol and then fractionated with n-hexane, ethyl acetate, and distilled water. The glucose-lowering effects of V. sinaiticum at 100, 200, and 400mg/kg were then studied. Glibenclamide was used as a positive control at a dose of 5 mg/kg. For oral glucose tolerance tests and hypoglycemia tests, Tween 2% was used as a negative control, while citrate buffer was used as a negative control for antihyperglycemic investigations. The results from the study were evaluated using one-way ANOVA, and then Tukey's post hoc multiple comparison test was performed. Results: Blood glucose levels were significantly reduced by the V. sinaiticum 80% methanol extract at 400 mg/kg (p<0.05). The blood glucose levels were significantly lowered by the aqueous residue at 400 mg/kg (p<0.05) and the ethyl acetate fractions at 200 mg/kg (p<0.01) and 400 mg/kg (p<0.001); however, none of the fraction extracts resulted in hypoglycemic shock in healthy mice. Higher glucose tolerance was seen in orally glucose-loaded mice after exposure to 80% methanol extracts at 200 and 400 mg/kg (p<0.05), the aqueous residual fraction at 200 mg/kg (p<0.01), and the ethyl acetate fraction at 200 and 400 mg/kg (p<0.05). The ethyl acetate fraction at 200 and 400 mg/kg (p<0.01), the 80% methanol extract at 400 mg/kg (p<0.05) and the aqueous residue at 400 mg/kg (p 0.01) significantly lowered blood glucose levels in streptozotocin-induced diabetic mice. Conclusion: The results of this study revealed that the 80% methanol extract and solvent fractions of V. sinaiticum Benth leaves are endowed with antidiabetic activity.

Anti-Inflammatory and Analgesic Activity of Methanolic Root Extract of Verbascum sinaiticum Benth.

Introduction: Pain in its various forms is undoubtedly the most common ailment known to human beings. Non-steroidal anti-inflammatory drugs (NSAIDs) and opioid analgesics are widely used to treat pain. However, long-term use of NSAIDs and opioids causes serious adverse effects on various organs. As a result, looking for drugs with better efficacy and lesser adverse effects appears crucial. For this purpose the obvious search begins from traditional medicines, particularly herbs. Therefore, this study investigated analgesic and anti- inflammatory activity of 80% methanol root extract of Verbasicum sinaiticum Benth (VS) in vivo. Methods: The dried and crushed plant material was macerated with 80% methanol sequentially and dried with lyophilizer. As per the acute toxicity study conducted elsewhere, 100 mg/kg, 200 mg/kg and 400 mg/kg doses of extract were used in the acetic acid induced writhing, hot plate test, as well as carrageenan and formalin induced anti-inflammatory models. As a positive control, aspirin 150 mg/kg was used for anti-nociceptive and anti-inflammatory model and morphine 10 mg/kg was used for central analgesic models. Results: VS200 and VS400 doses of the extract significantly (p< 0.05) reduced acetic acid induced writhing as compared with the control group. Similarly in hot plate test also, both VS200 and VS400 groups demonstrated significant (p< 0.05 at 30 min and p< 0.001 at 60 and 120 min) analgesic effect in comparison with the control and VS100 groups. Furthermore, in carrageenan and formalin induced anti-inflammatory test both VS200 and VS400 were shown to produce significant (p< 0.05) anti-inflammatory effect at the later hours and days. Conclusion: The findings from this study suggest that 80% methanol root extract of V. sinaiticum possesses peripheral and central analgesic as well as anti-inflammatory activity, possibly emanating from the phytochemicals present in the hydroalcoholic crude extract.

Antitrypanosomal activity of Verbascum sinaiticum Benth. (Scrophulariaceae) against Trypanosoma congolense isolates.

BACKGROUND: African Trypanosomiasis is a neglected tropical disease with a large impact on the livelihood of the rural poor in Sub-Saharan Africa. The available drugs for managing this disease are old, expensive and are facing the problem of drug resistance. Thus, the aim of this study was to evaluate in vivo antitrypanosomal efficacy of aqueous and absolute methanol leaf extracts of Verbascum sinaiticum Benth. against Trypanosoma congolense field isolate. METHODS: Verbascum sinaiticum (Local name 'qetetina') is a biennial plant, and 60-150 cm tall. It is traditionally used to treat wound, stomachache, viral infection, cancer, sunstroke, fever, abdominal colic, diarrhea, hemorrhage, anthrax, and hepatitis. The efficacy of aqueous and absolute methanol leaf extracts of V. sinaiticum was evaluated in a randomized experiment with Swiss albino mice infected with T. congolense field isolate. The extracts were administered at doses of 100, 200 and 400 mg/kg by intraperitoneal injection for seven days at 12 Days Post-Infection (DPI) when the peak parasitaemia level was approximately 10(8) trypanosomes/ml. Parasitaemia, Packed Cell Volume (PCV), mean survival time and change in body weight were used as indices for monitoring the efficacy of the extracts. Diminazene (28 mg/kg) was used as a positive control while 2 % Tween was used as the negative control. Phytochemicals screening were conducted following standard methods. RESULTS: The extracts showed no toxicity effect in Swiss albino mice and had LD50 above 2000 mg/kg. The phytochemicals screened in V. sinaiticum were alkaloids, flavonoids, glycoside, saponins, steroids, phenolic compounds, and tannins. The mice treated with absolute methanol leaf extract of V. sinaiticum at 400 mg/kg dose had significantly lower mean parasitaemia (7.20 ± 0.16) (p < 0.001) as compared to the negative control group (8.82 ± 0.12) on day 14 of treatment. Animals treated with the same dose had significant (p < 0.001) higher PCV value and body weight and as well as the highest mean survival time of 40.20 ± 0.31 days as compared to the negative control at the end of the observation period. CONCLUSION: This study established that Verbascum sinaiticum had trypanocidal activity. The crude extracts have partially eliminated trypanosomes in a dose-dependent manner. The study can be a basis for future in-depth analysis of the biologically active chemicals.