Quantification Of Total Flavonoids Research Articles

UPLC-MS/MS analysis and chemical profile of Ficus pertusa L. bark, an ethnomedicinal resource from intertropical America.

Background Ficus pertusa is a native species from intertropical America used for medicinal purpouses. However, despite its importance as ethnobotanical resource, there is poor chemical information about its extracts. Therefore, the objective of this study was to evaluate the phytochemical profile of the hydroalcoholic extract of F. pertusa barks through UPLC-MS/MS, as well as its phenolic and flavonoid content. Methodology The chemical identification was performed by Ultra-High Liquid Chromatography coupled to Mass Spectrometry, whereas the Total Phenolic content and Total Flavonoid quantification were measured through UV spectrometry. Results As a result, 19 substances were identified, which are mostly included within the group of condensed tannins, phenolic acids and flavonoids, such as procyanidins, chlorogenic acid and luteolin. Conclusion The identification of the main substances of F. pertusa’s bark and the measurement of its polyphenols content can guide the understanding of the pharmacological mechanisms involved in its ethnomedicinal uses.

Antioxidant and Hepatoprotective Activity of an Extract from the Overground Parts of Phlomis russeliana Lag. ex Benth

An evaluation of the possible hepatotoxicity/hepatoprotective effects of a defatted extract of the above ground parts of Phlomis russeliana was conducted in vitro and in vivo. The extract was tested in vitro on hepatocytes alone and in a carbon tetrachloride (CCl4)-bioactivation model. The same toxic substance was used for the in vivo evaluation on old Wistar rats. The extract was standardised via high performance liquid chromatography (HPLC) by the quantification of total flavonoids and verbascoside. Gallic acid equivalents were used to express the content of total phenolic compounds. The identification of flavonoids in this species was undertaken for the first time. The extract was not statistically hepatotoxic in vitro on the isolated rat hepatocytes. In the CCl4-induced hepatotoxicity model, the extract had a hepatoprotective effect, which was concentration-dependant (the highest at 50 µg/mL). An in vivo study on old rats confirmed the observed antioxidant and hepatoprotective effects. The histological findings were favourable for the rats, given the extract and CCl4 in combination. They had an unchanged organ structure, which is commensurable with these animals, treated with a combination of CCl4 and silymarin.

Antioxidant, antimicrobial and anticholinesterase activity of Dittrichia graveolens

Plants are responsible for many different biological activities. In our study, antioxidant, antimicrobial and anticholinestrase activities of Dittrichia graveolens (L.) Greuter were determined. In addition, the total phenolic and flavonoid contents of the plant were measured. The aerial parts of the plant were extracted with ethanol. Antioxidant activities of the extracts were measured with Rel assay kits. Antimicrobial activity was determined using the agar dilution method. Anticholinesterase activity was determined by measuring acetyl and butyrylcholinesterase activities. Determination of total phenolic content was determined using the Folin-Ciocalteu reagent. Total flavonoid quantification was performed using aluminum chloride assay. As a result of the study, the TAS value of the plant was determined as 6.933±0.121, the TOS value was 12.535±0.244 and the OSI value was 0.181±0.006. It was determined that the plant extract was effective against microorganisms at concentrations of 50-400 µg/mL. Additionally, the anti-AChE value was found to be 25.88±1.73 and the anti-BChE value was 45.32±2.26. Total phenolic content was determined as 86.42±2.72, and total flavonoid content was determined as 117.96±1.93. As a result, it was determined that D. graveolens has antioxidant, antimicrobial and anticholinesterase activity.

Formulation of Vitex negundo L. (Verbenaceae) leaf used in Traditional Medicine Hospital of Thua Thien Hue and evaluation of the product quality

Background: Vitex negundo L. leaves are a medicinal herb with many therapeutic effects, especially bone and joint diseases. At Traditional Medicine Hospital of Thua Thien Hue, method of using Vitex negundo L. leaves for patients who have joint diseases was shown to good treatment results. Besides, preparation of cream are convenient to use and take advantage of a precious and available medicinal herb of Vietnam. Objectives: To determine the formula, preparation process of products from the leaves of Vitex negundo L., determining the total flavonoid content of the product obtained and initial investigation of anti-inflammatory effects of prepared products. Methods: Building oriented formulas and investigating excipients. Development of a preparation process by emulsification method. Evaluation of the quality of prepared products by the Center for Drug - Cosmetic - Food Testing in Thua Thien Hue. Quantification of total flavonoids according to the method of Chang (2002). Investigation of anti-inflammatory effects based on carrageenan inflammation model in experimental rats. Results: Cream formula with the main ingredient was condensed glue of Vitex negundo L. 5%; Olive oil 10%; Glycerin 4%; Propylene glycol 15%; Ceteareth 20,4%; Cremophor RH40 7%; Nipagin 0.18%; Nipazol 0.02%; Menthol 0.05%. The cream product was smooth, homogeneous, dark brown in color, lightly scented, and met the basic standards. Total flavonoid content condensed glue of Vitex negundo L. was 13.18 ± 0.207, cream of Vitex negundo L. was 12.96 ± 0.163 (p > 0.05). The product was preliminarily evaluated to have the anti-inflamation effect in mice. Conclusions: Building a complete cream formula that met quality standards, the total flavonoid content was not lost much after the preparation process Key words: Vitex negundo L., the total flavonoid content, quality standards.

Costus spiralis extract restores kidney function in cisplatin-induced nephrotoxicity model: Ethnopharmacological use, chemical and toxicological investigation

Ethnopharmacological relevanceCostus spiralis (Jacq.). Roscoe (Costaceae) is traditionally used in Brazil for the treatment of kidney diseases such as pyelonephritis, urethra inflammation, kidney stones, and inflammatory conditions. There are reports of its use by Brazilian Indians since the 17th century when it was known as “pacocatinga.” Currently, the use of the Costus species in Brazil is widespread, which was evidenced by the inclusion of the genus in the Brazilian National List of Medicinal Plants of Interest to the Unified Health System (RENISUS). Aim of the studyThis study aimed to confirm the ethnopharmacological use of Costus spiralis in the treatment of kidney diseases, toxicity study using animal models, and the phytochemistry of the species. Materials and methodsThe chemical profile of Costus spiralis leaves extract (CSLE) was obtained for the hydroethanolic extract by ultra-performance liquid chromatography coupled to a mass spectrometer and ultraviolet detector with diode array (UPLC-UV/DAD-ESI-MS). The acute oral toxicity of the extract was predicted using the neutral red uptake cytotoxicity assay. Wistar rats were used in a model in vivo for confirmation of acute oral toxicity (2000 mg/kg p.o. for 14 days.) and determination of the effect on a cisplatin-induced nephrotoxicity model. ResultsThe analysis by UPLC-UV/DAD-ESI-MS showed that the chemical composition of the extract is mostly di-glycosylated flavones of apigenin. In the extract were identified the flavones vicenin II and schaftoside. The quantification of total flavonoids by spectrometry showed 0.880%. CSLE proved to be safe for acute oral administration (2000 mg/kg) with an IC50 value of 222.9 μg/mL and predicted oral toxic dose of 523.82 μg/mL in a neutral red uptake cytotoxicity assay. The absence of death allows the classification of the extract in class 5 according to OECD 423 guidelines and therefore it can be considered as a high acute safety product, which is highly relevant, considering the wide popular use of the species. In the cisplatin-induced nephrotoxicity model, C. spiralis extract (5, 15, and 30 mg/kg) significantly improved renal function, reversing almost completely the effects on plasma creatinine levels and creatinine clearance (p < 0.001). ConclusionsThis study demonstrates that oral administration of Costus spiralis extract leaves is safe and effective in restoring the renal function in rats in a cisplatin-induced nephrotoxicity. It is suggested that the observed activity is related to the flavonoids present. This hypothesis should be confirmed, and the participation of other secondary metabolites should be investigated in the future.

Determination and validation of spectrophotometric analytical method for quantification of total flavonoids in the leaves of Azadirachta indica A. Juss. (Meliacease) and optimization of the ultrasound-assisted extraction conditions

Azadirachta indica A. Juss. (Meliaceae) is a plant whose leaves present several pharmacological activities, among them: antioxidant, antimicrobial and hypoglycemic. The present work aims to determine and validate a spectrophotometric analytical method for quantification of total flavonoids, expressed as rutin, in A. indica leaves and to optimize the extraction conditions of these flavonoids. For the selection of the spectrophotometric method, four analytical methods were investigated. The method selected as the most efficient, simple, fast and sustainable has been validated and optimized. The validation was performed according to criteria of national and international regulatory agencies. Box Behnken design (33) and the response surface methodology (RSM) were used to estimate the best extraction conditions. The analytical method that met the study criteria consisted of ultrasound-assisted extraction (UAE) of the plant material in hydroethanolic solution, at a temperature of 60 ºC, for 30 minutes, followed by filtration, dilution of the extract in ethanol/0.02 M acetic acid solution (99:1) and absorbance reading in a spectrophotometer at 364nm. In the validation stages, this method was shown to be linear (linear interval - 20 to 80 μg/mL), precise (intraday and interday), accurate (mean recovery of 120.8%), robust and selective (absence of matrix effect). The best conditions for the UAE process were: Ethanol content 30% (p/p), plant material/solvent ratio 0.2 g/mL and extraction time of 33 minutes. The spectrophotometric method and the UAE conditions determined in this study are suitable for use in the quality control of plant drugs and ethanol extracts from the leaves of Azadirachta indica.

Development and validation of analytical method by spectrophotometry UV-Vis for quantification of flavonoids in leaves of Senna occidentalis Link

Senna occidentalis Link (Fabaceae), known as “fedegoso”, is used in folk medicine due to its anti-inflammatory, antioxidant, muscle relaxant and inhibiting lipid peroxidation imputed to flavonoids contained in its composition. The species is also a constituent of processed products are commercialized in various parts of the world including Brazil, although there are few reports in the literature about the development of an analytical method for quantification of flavonoids in it. The main purpose of this research was the evaluation of an analytical methodology to determinate the content of total flavonoids in leaves of S. occidentalis, for quality control. The parameters evaluated were: drug: solvent ratio, concentration of aluminum chloride and reaction time. The quantification of total flavonoids was obtained after reaction with aluminum chloride by UV/Vis spectrophotometry. The results revealed a maximum absorption peak (λ = 392 nm) was the amount of 0.5 g of a plant raw in 100 mL of solvent and there were no significant influences between the concentrations of aluminum chloride or hydroalcoholic solutions analyzed. The evaluation of reaction time allowed to characterize the kinetics as slow, stabilizing from 60 min, choosing this as reading time. The method was specific, linear, precise, accurate and robust, according to the specifications set in RDC n. 166/2017. Finally, the results of the study showed that the measured methodology is simple and accurate and can be applied for quality assessment of the species S. occidentalis.

Evaluation of 2,2-diphenyl-1-picrylhydrazyl, secondary metabolite contents and antimicrobial efficacy of Blastonia garcinii(Burm.f.) Cogn.

This study emphasizes screening of DPPH, secondary metabolites and antimicrobial activity of Blastonia garcinii an annual wine with a slender stem, climbing using tendrils, leaves are ovate and flowers are unisexual. The leaves were collected, shade dried and powdered. The powdered sample was extracted by using soxhlet apparatus with petroleum ether and ethanol. The antioxidant activity of the plant extracts was measured using spectrophotometry by their ability to scavenge free radicals such as DPPH (2,2-diphenyl-1-picrylhydrazyl), hydrogen peroxide and also quantification of total flavonoids, total phenol content and antimicrobial activity of plant extracts. The significant result observed in when compared Petroleum ether and Ethanol extract, the DPPH and H 2 O 2 activities and the total phenol and total flavonoid content and Antibacterial and antifungal activity were shown significant activity in ethanolic extract of B. garcinii.

Phytochemical Evaluation, Embryotoxicity, and Teratogenic Effects of Curcuma longa Extract on Zebrafish (Danio rerio).

Curcuma longa L. is a rhizome plant often used as traditional medicinal preparations in Southeast Asia. The dried powder is commonly known as cure-all herbal medicine with a wider spectrum of pharmaceutical activities. In spite of the widely reported therapeutic applications of C. longa, research on its safety and teratogenic effects on zebrafish embryos and larvae is still limited. Hence, this research aimed to assess the toxicity of C. longa extract on zebrafish. Using a reflux flask, methanol extract of C. longa was extracted and the identification and quantification of total flavonoids were carried out with HPLC. Twelve fertilized embryos were selected to test the embryotoxicity and teratogenicity at different concentration points. The embryos were exposed to the extract in the E3M medium while the control was only exposed to E3M and different developmental endpoints were recorded with the therapeutic index calculated using the ratio of LC50/EC50. C. longa extract was detected to be highly rich in flavonoids with catechin, epicatechin, and naringenin as the 3 most abundant with concentrations of 3,531.34, 688.70, and 523.83μg/mL, respectively. The toxicity effects were discovered to be dose-dependent at dosage above 62.50μg/mL, while, at 125.0μg/mL, mortality of embryos was observed and physical body deformities of larvae were recorded among the hatched embryos at higher concentrations. Teratogenic effect of the extract was severe at higher concentrations producing physical body deformities such as kink tail, bend trunk, and enlarged yolk sac edema. Finally, the therapeutic index (TI) values calculated were approximately the same for different concentration points tested. Overall, the result revealed that plants having therapeutic potential could also pose threats when consumed at higher doses especially on the embryos. Therefore, detailed toxicity analysis should be carried out on medicinal plants to ascertain their safety on the embryos and its development.

Phenolic compounds, carotenoids, and antioxidant capacities of a thermo-tolerant Scenedesmus sp. (Chlorophyta) extracted with different solvents

The human body can fight against the adverse effects of chronic exposure to environmental pollutants and stress by consumption of a diet rich in antioxidants. Although fruits and vegetables are the predominant sources, alternative sources of antioxidants such as microalgae are also being explored. Here, we investigate the antioxidant capacity, total phenolic, flavonoid, and carotenoid contents of novel thermo-resistant green microalga Scenedesmus sp. ME02. This strain has previously been shown to have a high polyunsaturated fatty acid content. Four different solvents were used for extraction and the antioxidant capacity was determined to be 3.71 ± 0.11 and 47.01 ± 3.14 μmol Trolox eq. g−1 DW in ethanol/water mixture by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays, respectively. Total phenolic, flavonoid, and carotenoid contents in ethanol/water were measured as 5.40 ± 0.28 mg gallic acid eq. g−1 DW, 1.61 ± 0.76 mg quercetin eq. g−1 DW, 0.61 ± 0.05 mg g−1, respectively. This is one of the few studies that reports the presence and quantification of total flavonoids in microalgae. The correlation between the total phenolic content and FRAP assay, but not the DPPH assay, was statistically significant. Finally, 12 different phenolic compounds were analyzed by reverse-phase HPLC and ethyl acetate extract showed substantial amounts of quercetin (0.84 ± 0.12 mg g−1 DW) and rutin (0.11 ± 0.08 mg g−1 DW). Quercetin amount was also high in the ethanol/water extract along with gallic acid, 4-hydroxy benzoic acid, and chlorogenic acid. To the best of our knowledge, this is the first study that reports significant amounts of quercetin and rutin in a microalgal species.

Antioxidant effect of Morus nigra on Chagas disease progression.

ABSTRACTConsidering the widespread popular use of Morus nigra and the amount of scientific information on its antioxidant and anti-inflammatory activity, the effectiveness of this phytotherapeutic compound in the parasitemia progression during the acute phase of Chagas disease and its role in the development of the inflammatory process as well as its effects on the oxidative damage in the chronic phase of infection were evaluated. Thus, 96 male Swiss mice were randomly divided into eight groups, four groups were uninfected controls, and four groups were intraperitoneally infected with 5.0 x 104 blood trypomastigotes forms of T. cruzi QM2 strain. Four batches composed of one uninfected and one infected group were respectively treated with 70% alcohol solution and 25 μL, 50 μL and 75 μL of the phytotherapeutic compound. Levels of antioxidant elements (TBARS, FRAP, GSH and Sulfhydryl groups) were measured in plasma samples. The phytotherapeutic compound’s antioxidant activity was measured by polyphenol and total flavonoid quantification, DPPH, NO, and FRAP method. Our results showed that the vehicle influenced some of the results that may have physiological relevance in Chagas disease. However, an important action of M. nigra tincture was observed in the progression of Chagas disease, since our results demonstrated a reduction in parasitemia of treated groups when compared to controls, especially in the group receiving 25 µL. However, in the chronic phase, the 50-µL dosage presented a better activity on some antioxidant defenses and minimized the tissue inflammatory process. Results indicated an important action of M. nigra tincture on the Chagas disease progression.

Phytochemical analysis and salivary amylase inhibition activities of Carica papaya leaf and Garcinia mangostana pericarp extracts and partially purified fractions

Plant-derived digestive enzyme inhibitors particularly those targeted to carbohydrate metabolism has been the focus of recent studies as natural supplements for weight control and diabetes. The present study explores the salivary amylase inhibition activity of Garcinia mangostana (Linn.) pericarp extracts and Carica papaya (Linn.) leaf extracts and fractions, as well as perform phytochemical screening and quantification, and thin layer – and high performance liquid chromatographic profiling. ­Results show that crude extracts and purified fractions were able to inhibit salivary amylase, with C. papaya fraction 1 being the most active at 30.89% inhibition. Phytochemical screening of all extracts tested ­positive for tannins, glycosides, phenolics, flavonoids and alkaloids. Quantification of phenolics showed that extracts contained high levels of phenolics, with C. papaya crude extract having the highest content with 219.0±12.7 mg GAE/g extract followed by G. mangostana crude extract with 247.1±18.0 mg GAE/g extract. Quantification of total flavonoids also showed C. papaya crude extract to contain the highest content with 55.12±0.679 mg QE/g extract. All extracts contained negligible alkaloid content, though. HPLC and TLC profiling showed several peaks and bands, when viewed in 210 nm and UV light, respectively. These results demonstrate in vitro the salivary amylase inhibitory activity of both plants and their potential as antidiabetic drug candidates; however, further studies need to be done, like isolation and structure elucidation of active components and toxicity assays. Keywords: Amylase inhibition, phytochemical quantification, Carica papaya, Garcinia mangostana

Comparative cytotoxic and spectrophotometric quantification of phytochemicals of the methanol extracts of the leaf and root bark of &lt;i&gt;Securinega virosa&lt;/i&gt; (Roxb ex. Willd) Baill (Euphorbiaceae)

The comparative cytotoxic and spectrophotometric quantification of phytochemicals of the methanol extracts of the leaf and root bark of Securinega virosa was carried out. Phytochemical screening and spectrophotometric quantification of total flavonoids and phenolics of the extracts were carried out using standard reported methods. Phytochemicals detected in the extracts include alkaloids, cardiac glycosides, flavonoids, steroids, tannins, terpenoids and saponins. The total flavonoids and phenolic contents were observed to be more on the leaves than the root bark. The cytotoxic effects of the methanol extracts were evaluated between 10-400 µg/ml over a period of 24 hr using the tadpole mortality assay. At 100 µg/ml, the methanol extract of the leaf produced 73.30 ± 3.33% mortality which increases to 100 % at 200 and 400 µg/ml while the root barks produced 26.78 ± 4.45 and 56.70 ± 6.67 % mortalities respectively at the same concentrations. The significant cytotoxic effects observed coupled with the presence of phytochemicals such as phenolics supports the ethnomedicinal claim of this plant in treating cancer in Ogun State. Keywords: Cytotoxicity; Phytochemicals, Securinega virosa ; Phenolics; Raniceps raninus; Spectrophotometric

Hypotensive effect and endothelium-dependent vascular action of leaves of Alpinia purpurata (Vieill) K. Schum

The aims of this study were to evaluate the chemical profile, vascular reactivity, and acute hypotensive effect (AHE) of the ethanolic extract of leaves of Alpinia purpurata (Vieill) K. Schum (EEAP). Its chemical profile was evaluated using HPLC-UV, ICP-OES, and colorimetric quantification of total flavonoids and polyphenols. The vascular reactivity of the extract was determined using the mesenteric bed isolated from WKY. AHE dose-response curves were obtained for both EEAP and inorganic material isolated from AP (IAP) in WKY and SHR animals. Cytotoxic and mutagenic safety levels were determined by the micronucleus test. Rutin-like flavonoids were quantified in the EEAP (1.8 ± 0.03%), and the total flavonoid and polyphenol ratios were 4.1 ± 1.8% and 5.1 ± 0.3%, respectively. We observed that the vasodilation action of EEAP was partially mediated by nitric oxide (·NO). The IAP showed the presence of calcium (137.76 ± 4.08 μg mg-1). The EEAP and IAP showed an AHE in WKY and SHR animals. EEAP did not have cytotoxic effects or cause chromosomic alterations. The AHE shown by EEAP could result from its endothelium-dependent vascular action. Rutin-like flavonoids, among other polyphenols, could contribute to these biological activities, and the calcium present in EEAP could act in a synergistic way.

Quantification of Total Flavonoids, Total Phenolic Compounds and Antioxidant Activity of Extracts from the Seed of &lt;i&gt;Viburnum odoratissimum&lt;/i&gt;

The fruit of Viburnum odoratissimum is an elliptical berry. The aims of our study were the quantification of some antioxidants using colorimetric assays and the determination of antioxidant activities by three methods, and the relationships between these bioactive compounds and antioxidant activities of the crude extract (CE) and its four fractions derived from the seed of the fruit of Viburnum odoratissimum were investigated. The results showed that the n-butanol-soluble fraction (BF)was the richest fraction in total flavonoids and total phenolic. The ethyl acetate-soluble fraction (EAF) and BF showed good superoxide radical scavenging activity(SRSA) and reducing power(RP), while the petroleum ether-soluble fraction(PEF) possessed the highest inhibiting lipid peroxidantion activity(LPA).There were statistically significant correlations between some antioxidants and antioxidant activities.

Quantification of Total Flavonoids and Phenolic Acids from Microwave Irradiated and Non-irradiated Plants

Quantification of Total Flavonoids and Phenolic Acids from Microwave Irradiated and Non-irradiated Plants

Geopropolis from Melipona scutellaris decreases the mechanical inflammatory hypernociception by inhibiting the production of IL-1β and TNF-α

Ethnopharmacological relevanceThe pharmacological activity of geopropolis collected by stingless bees (important and threatened pollinators), a product widely used in folk medicine by several communities in Brazil, especially in the Northeast Region, needs to be studied. ObjectiveThe aim of this study was to evaluate the antinociceptive activity of Melipona scutellaris geopropolis (stingless bee) using different models of nociception. Material and methodsThe antinociceptive activity of the ethanolic extract of geopropolis (EEGP) and fractions was evaluated using writhing induced by acetic acid, formalin test, carrageenan-induced hypernociception, and quantification of IL-1β and TNF-α. The chemical composition was assessed by quantification of total flavonoids and phenolic compounds. ResultsEEGP and its hexane and aqueous fractions showed antinociceptive activity. Both EEGP and its aqueous fraction presented activity in the mechanical inflammatory hypernociception induced by the carrageenan model, an effect mediated by the inhibition of IL-1β and TNF-α. The chemical composition of EEGP and its hexane and aqueous fractions showed a significant presence of phenolic compounds and absence of flavonoids. ConclusionOur data indicate that geopropolis is a natural source of bioactive substances with promising antinociceptive activity.

Optimization of Extraction Condition and Quantification of Total Flavonoids in Elaeagni Folium

Introduction: Elaeagni Folium has been used as a crude drug to cure lung deficiency cough, asthma and shortness of breath for a long time in China. The research aimed to optimize the extraction condition and measure the amount of total flavonoids in this medicinal material.Methods: Reflux extraction was an efficient method, compared with sonication and soxhlet extraction, to extract total flavonoids. On the basis of single factor test, the L9(34) orthogonal test was used in the optimization of technological parameters by investigating the ethanol concentration, extraction time and the material/solvent ratios. Using 0.1 mol·L− aluminum chloride (AlCl3) solution as chromogenic agent, the total flavonoids content was measured by ultraviolet and visible spectrophotometry.Results: The optimal extraction parameters showed that: ethanol concentration was 85%, ratio of solvent to raw material 40:1 and extraction time 1.5 h. The amount of total flavonoids in the leaf of Elaeagnus pungens Thunb. collected in different harvest time and habitats was 20.8–33.2mg·g-1.Conclusions: It indicated that the amount of total flavonoids had significant difference in this crude drug from different habitats but at the same time, while the total flavonoids content was stable in the samples collected in different harvest time but at the same habitat.