Resumo em Inglês:

ABSTRACT Digestive herbal teas in the form of monoherbal infusions or in mixtures are the most commonly used form of phytotherapy in Bahía Blanca (Argentina). This practice is based on family tradition reinforced by current trends reverting to the ingestion of natural products. The objective of this work was to analyze the botanical quality of the digestive mixtures sold in Bahía Blanca as fine cut mixtures (in tea bags) and thick cut mixtures (fragmented herbs). Samples of seven mixtures sold in the form of tea bags and four as bags of fragmented herbs were studied under stereoscopic and optical microscopes and micrographic studies were carried out following conventional protocols. Observed characteristics were compared with those in the reference bibliography for verified genuine monoherbs. In the studied products a total of 21 different species were found. There was total concordance between the species declared on the label and those identified in the samples in six of the mixtures in the form of tea bags and in only one of the bags of fragmented herbs. Adulterations were found in four of the samples analyzed (one in a tea bag and three in the fragmented mixtures). The labels on only two of the tea bag samples presented complete and correct information; in the remaining samples irregularities were found related to: the taxonomic identification of the species, the absence of a lot number, date of expiry, the plant parts used and proportions of the herbs etc. The findings underline the great importance of quality controls as a tool to protect the rights of phytotherapy consumers.

Resumo em Inglês:

ABSTRACT Bananas and plantains are herbaceous monocotyledonous plants belonging to the genus Musa, Musaceae, which has a widespread distribution around the world. Various parts of banana plant are commonly used in traditional medicines. Several species of Musa are reported to possess anti-inflammatory, anti-hyperglycemic and antidiabetic properties. This work is aimed at studying the morphological and anatomical characteristics of the inflorescences of Musa × paradisiaca L., that could contribute to the characterization of these species cultivated in Brazil. Plant materials were collected and prepared in accordance with standard optical microscopy techniques. Morphological characterizations were conducted using morphological descriptors for inflorescences, including some descriptors from International Plant Genetic Resources Institute for Musa spp. Microscope slides were prepared using glycol-methacrylate and were stained in toluidine blue. Main features observed for M. × paradisiaca inflorescence were amphistomatic bracts with tetracytic stomata, fiber caps next to the phloem, adaxial and abaxial uniseriate epidermis, and papillose on the abaxial face. Outer tepals have multilayer epidermis and vascular bundles aligned next to the abaxial face. Free tepal has unilayeredepidermis. Anthers are tetrasporangiate and the locules are separated by the septum. Ovary is inferior and trilocular with external unilayered and internal epidermis. The main morpho-anatomical characteristics of inflorescence of Musa × paradisiaca are highlighted in this study, contributing to provide more information about the characterization of this species cultivated in Brazil.

Resumo em Inglês:

ABSTRACT The root of Anacyclus pyrethrum (L.) Lag., Asteraceae, is very widely used for treating various diseases in Traditional Uygur Medicine, particularly in the treatment of vitiligo. However, there have been few studies on the quality standards of A. pyrethrum in China. A. pyrethrum contains abundant N-alkylamides, which are considered to be the principal components. Therefore, based on the previous research in our group, six N-alkylamides were obtained by using column chromatography. We used ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry to determine the mass spectrometry cleavage mechanism of these six monomer components and established the mass spectrometry cleavage law of N-alkylamides. Then, we used the ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry method to rapidly identify and analyze the N-alkylamide components of the A. pyrethrum methanol extract. Finally, twenty N-alkylamides were identified, including eleven N-isobutylamides, two N-methyl isobutylamides, six 4-hydroxyphenylethyl-amide and one 2-phenylethylamide. Five of these compounds were identified as new compounds that have not been reported to date. Two of these compounds were identified for the first time in this herb. Therefore, this work provides an approach for the quality analysis of N-alkylamides in the root of A. pyrethrum. A search of the literature showed that the content determination in the A. pyrethrum quality standard is still a remaining problem. N-alkylamides are the main components of A. pyrethrum. Even though ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry has the advantages of lower time and higher efficiency compared to high-performance liquid chromatography, considering the ease of repeatability and universality of the quality control method, we chose to use high-performance liquid chromatography for content determination. In this experiment, high-performance liquid chromatography was used for the first time to establish a simple, rapid and accurate method for evaluating the N-alkylamide content in A. pyrethrum with five N-alkylamides used as the standards. Finally, this work provides a qualitative and quantitative method for the analysis of N-alkylamides in A. pyrethrum, improving the quality control standards for A. pyrethrum.

Resumo em Inglês:

ABSTRACT In this work, we developed and validated a HPLC-PDA method for the quantification of hibalactone in Hydrocotyle umbellata L., Araliaceae, subterraneous parts extracts and optimized its ultrasound-assisted extraction. Chromatographic separations were carried out with an isocratic mobile phase of acetonitrile/methanol/water (10:65:25), a flow of 0.8 ml min−1, detection at 290 nm and C18 column (250 × 4.6 mm, 5 µm). The method validation parameters were determined according to Brazilian legislation. The optimization of the hibalactone ultrasound-assisted extraction was performed using Box–Behnken design and response surface methodology. The HPLC method for hibalactone quantification proved to be selective, linear, precise, accurate and robust, being useful for the analysis of hibalactone in H. umbellata subterraneous parts extracts. The optimal ultrasound-assisted extraction conditions were obtained with solid-to-liquid ratio of 1:5 g ml−1, ethanolic strength of 70% (v/v) and temperature of 65 °C. The results can provide support of the quality control and standardization of raw materials from H. umbellata.

Resumo em Inglês:

ABSTRACT A validated method for the identification and authentication of tingenone and pristimerin was developed using HPLC. The chromatographic profile analysis was combined with simultaneous quantification in Crossopetalum rhacoma Crantz, Cassine xylocarpa Vent, Semialarium mexicanum (Miers) Mennega (known as cancerina), and Maytenus phyllanthoides Benth, through microwave-assisted extraction. The HPLC profiles of the four analyzed species showed three similar signals, which corresponded to the main chemotaxonomic markers of the Celastraceae family: quinonemethide triterpenes. HPLC profile analysis was used as a tool to identify the relationship with the quinonemethide triterpenes, for establishing the taxonomic position of some species whose placement in, or within, the Celastraceae family is uncertain.

Resumo em Inglês:

ABSTRACT In Thai traditional medicine, Pluchea indica (L.) Less., Asteraceae, leaf has been widely used for the treatment of diabetes mellitus, tumors, hypertension, cystitis, and wounds. P. indica herbal tea is commercially available in Thailand as a health-promoting drink. The study was conducted to develop and validate a high-performance thin-layer chromatography (HPTLC) method for the quantitative analysis of chlorogenic acid, 3,4-O-dicaffeoylquinic acid, and 3,5-O-dicaffeoylquinic acid in P. indica leaf extract and their commercial products in Thailand. The method was validated according to ICH guidelines. The proposed HPTLC method showed acceptable validation parameters. The content of chlorogenic acid, 3,4-O-dicaffeoylquinic acid, and 3,5-O-dicaffeoylquinic acid in P. indica leaves from seven different provinces in Thailand was in the range of not detectable −1.94 ± 0.02%w/w, 0.71 ± 0.01–1.89 ± 0.05%w/w, and 1.00 ± 0.01–2.18 ± 0.03%w/w, respectively, while in the commercial products, it was in the range of 0.59 ± 0.03–2.17 ± 0.05%w/w, 0.53 ± 0.04–3.77 ± 0.03%w/w, and 0.88 ± 0.05–4.72 ± 0.10%w/w, respectively. The results indicated that plantation of P. indica in coastal saline land would be beneficial as it would increase the concentration of its active compounds and improve its medicinal quality. The developed high-performance thin-layer chromatography could be used as a rapid, reliable, less demanding, and cost-effective analytical method.

Resumo em Inglês:

ABSTRACT Acheflan® herbal medicine is the first medicine developed and produced in Brazil using Cordia verbenacea DC., Boraginaceae, essential oil as a constituent. C. verbenacea has anti-inflammatory properties, which have been directly related to α-humulene and trans-caryophyllene. Currently, the quantification of α-humulene and trans-caryophyllene in C. verbenacea has only been described by GC-MS. Although this technique is widely used it cannot be directly applied to finished medicinal products since they contain aqueous constituents. In this context, the objective of this work was to develop a methodology for quantifying C. verbenacea in raw materials and in pharmaceutical formulations using liquid chromatography. The Box–Behnken experimental design was used successfully to optimize the analytical method. The method developed in this study was validated and proven to be effective in the proper separation of α-humulene and trans-caryophyllene. In addition, the developed method was applied to commercial formulations (cream and aerosol) containing C. verbenacea essential oil, which resulted in very satisfactory separation without interference from excipients. C. verbenacea oil contains four times higher concentrations of trans-caryophyllene than α-humulene. Therefore, we propose that trans-caryophyllene can be used as a marker of C. verbenacea essential oil. Evaluation of trans-caryophyllene content would be especially valuable for applications where the concentrations are very low such as in permeation and release studies of dermatological formulations.

Resumo em Inglês:

ABSTRACT Natural products have been the most valuable source of chemical compounds in the discovery of novel medicines. Secondary metabolites from terrestrial and marine organisms have found considerable use in the treatment of numerous diseases and have been considered lead molecules both in their natural form and as templates for medicinal chemistry. Brazil has an exceptionally rich biodiversity, and a valuable source of secondary metabolites that can be useful for the development of bioproducts. Ipomoea species, Convolvulaceae, are mostly found in tropical and sub-tropical regions, including South America and many are used for nutritional and medicinal purposes. Ipomoea procumbens Mart. & Choisy is endemic from South America, and this is the first study reported on the chemical composition and biological activities of this species. The present work reports the tentatively identification of natural products present in the extracts using a high performance liquid chromatography–high resolution mass spectrometry method. Additionally, the antioxidant and antifungal biological activities of the leaves, roots and steams extracts and fractions of this species were evaluated. While for the antioxidant activity the hydromethanol fractions (leaves, stem and roots) were more active, the methanol fractions of leaves and stem provided better results for the antifungal assay.

Resumo em Inglês:

ABSTRACT Schisandra chinensis (Turcz.) Baill., Schisandraceae, is a well-known traditional Chinese medicine used mainly as a recipe for hepatoprotection. Modern researches have revealed that the hepatoprotection is related to its lignans and crude polysaccharide. In this study, we examined the effect and mechanism of S. chinensis total lignans on the liver injury induced by alcoholic. S. chinensis total lignans were extracted with ethanol extraction. The liver index, alanine aminotransferase and aspartate aminotransferase levels in serum of the rat culture supernatant were examined. The malondialdehyde level and superoxide dismutase activities in serum and liver tissue, and triacylglyceride content in liver tissue were tested. Western blot was conducted to determine cytochrome P450 2E1 expression in liver tissue of rats. The results showed that S. chinensis total lignans administration significantly inhibited alcohol-induced liver injury. In exploring the underlying mechanisms of S. chinensis total lignans action, we found that it significantly decreased Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), Glutamyl transpeptidase (γ-GT), reactive oxygen species (ROS) and malondialdehyde (MDA) level in livers/serum and inhibited the gene expression level of CYP2E1 in rat livers. The Nuclear factor erythroid-2 related factor 2 (Nrf2) gene expression and Nuclear factor erythroid-2 related factor 2 (Nrf2) protein nuclear transfer increased significantly, and significantly increased the expression of downstream target gene and protein heme oxygenase-1 (HO-1), Glutamate--cysteine ligase regulatory subunit (GCLM), NAD(P)H:quinine oxidoreductase 1 (NQO1). Moreover, S. chinensis total lignans decreased production of pro-inflammatory markers including Tumor Necrosis Factor-α (TNF-α), Interleukin-1beta (IL-1β) and Interleukin-6 (IL-6). In conclusion, these results suggested that the inhibition of alcohol-induced liver injury by S. chinensis total lignans is associated with its ability to inhibiting CYP2E1 activation and activating the Nrf2/Antioxidant response element(ARE) signaling pathway.

Resumo em Inglês:

ABSTRACT A new xanthone: mangostanaxanthone VIIII [1,3,5,6,7-pentahydroxy-2-(3-methylbut-2-enyl)-8-(3-hydroxy-3-methylbut-1-enyl) xanthone] (5) and four known xanthones: mangostanaxanthones I (1) and II (2), γ-mangostin (3), and mangostanaxanthone VII (4) were separated and characterized from the acetone fraction of Garcinia mangostana L., Clusiaceae (mangosteen) pericarps. Their structures were established based on various spectroscopic analyses in addition to HRMS and comparison with the literature. The α-amylase inhibitory potential of the isolated metabolites was evaluated. Compounds 1, 2, and 5 had the highest activity with % inhibition 72.5, 86.5, and 81.8, respectively compared to acarbose (97.1%, reference α-amylase inhibitor). The molecular docking study of the tested metabolites was estimated to shade up the rational explanation of the α-amylase inhibitory activity results. Moreover, the pharmacokinetic parameters were assessed using Swiss ADME. It is noteworthy that 1, 2, and 5 had similar binding poses as the X-ray crystal structure of acarbose, whereas the other metabolites possessed different binding mode that decreased their inhibitory capacity. Thus, these data reinforced the health benefit of mangosteen as an alternative medicine to help lowering the postprandial glucose absorption. Therefore, it could have a good potential for the treatment of diabetes.

Resumo em Inglês:

ABSTRACT The hepatoprotective activities of two traditionally used plants, Cleome droserifolia (Forssk.) Delile, Cleomaceae, and Artemisia annua L., Asteraceae, were recently reported. However, the biologically active metabolites responsible for this activity were not identified. The aqueous extract of C. droserifolia aerial parts, and the polar fraction of A. annua leaves were screened for their antioxidant activities using the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) assay. The in vitro viability of HepG-2 cells treated with CCl4 and the extracts were assessed by MTT assay. The effects of the extracts on the liver enzymes and the total soluble protein in CCl4-intoxicated HepG-2 cells were investigated. An HPLC/PDA/ESI/MS–MS based analysis was carried out for extract of C. droserifolia and polar fraction of A. annua. Both exhibited pronounced free radical scavenging activities (86 and 83%, respectively). Both showed a significant increase in cell viability: 86.43% for the extract of C. droserifolia and 79.32% for polar fraction of A. annua. Only the extract of C. droserifolia (39.6 ± 5.41 and 20.4 ± 6.91 IU/dl, respectively) and polar fraction of A. annua (40.8 ± 2.14 and 24.5 ± 3.11 IU/dl, respectively) restored the levels of liver enzymes (aspartate transaminase and alanine transaminase, respectively) compared to the CCl4 intoxicated group (87.5 ± 4.34 and 34.1 ± 8.12 IU/dl, respectively) and other herbal extracts. More than fifty phenolic secondary metabolites were identified in the extracts under investigation. The significant hepatoprotective activities of both extracts seemed to be strongly connected to their content of hydroxycinnamoyl quinic acids and flavonoids.

Resumo em Inglês:

ABSTRACT This study investigated the possible antioxidant and neuroprotective effects of alcesefoliside, isolated from Astragalus monspessulanus L., Fabaceae, against carbon tetrachloride (CCl4)-induced brain injury in Wistar rats. Iron sulphate/ascorbic acid lipid peroxidation was induced in rat brain microsomes and pre-incubated with alcesefoliside and silybin. Male rats were treated in vivo with alcesefoliside and with silymarin alone; animals challenged with CCl4; and pre-treated with alcesefoliside or silymarin in respective doses for 7 days, challenged with CCl4, followed by curative treatment (additional 14 days). The activity of acetylcholine esterase and the antioxidant enzymes: superoxide-dismutase, catalase, glutathione-peroxidase, glutathione reductase and glutathione-S-transferase as well as the biomarkers of oxidative stress malondialdehyde and reduced glutathione were measured. The alcesefoliside pre-treatment and consecutive curative treatment normalizes the activity of the antioxidant enzymes as well as levels of malondialdehyde and reduced glutathione. The observed effects on tissue level correlate with the histopathological observations of the brain. They were comparable to the effects of silymarin, used as a positive control. The results showed that alcesefoliside has a neuroprotective effect against CCl4-induced brain toxicity in rats.

Resumo em Inglês:

ABSTRACT Croton zehntneri Pax & K. Hoffm., Euphorbiaceae, or “canela-de-cunhé” is used in the Northeast Brazil to treat several diseases. Leaves and aerial parts of C. zehntneri are rich in volatile oil of high potential therapeutic. This study aimed to investigate volatile oil systemic toxicity after per oral treatment in rats. Volatile oil characterization (gas chromatography and mass spectrometry) showed 85.7% anethole and 4.8% estragole. Male Wistar rats (116–149 g) were treated with volatile oil (250 mg/kg p.o.) during ten weeks and evaluated for the following parameters: survival; food and water intake; body mass; absolute/relative organs weight; hemogram; plasma biochemical dosage; organs morphology. Volatile oil did not alter animal water and food consumption or the relative/absolute weight of most organs, but animals gained less weight. Volatile oil did not alter function biomarkers of pancreas, kidney, heart or liver, but increased plasma gamma-glutamyltranspeptidase (liver biomarker) and decreased uric acid (kidney biomarker). Although volatile oil had caused discrete morphological alterations in some organs, it did not induce architectural changes in these organs. In conclusion, the sub-acute per oral treatment with volatile oil no longer than ten weeks in rats offers small toxicity at doses below 250 mg/kg.

Resumo em Inglês:

ABSTRACT Gentamicin induced renal complications are well known in humans and animals. Medicinal properties of Withania somnifera (L.) Dunal, Solanaceae, are recognized to improve renal functions. However, the pharmacological function of W. somnifera is not completely understood. We sought to unravel medicinal therapeutic function of W. somnifera on gentamicin-induced nephrotoxicity in wistar rats. Twenty-four adult male wistar rats evenly divided into four groups to evaluate in vivo nephroprotective and nephrocurative function of W. somnifera in gentamicin induced nephrotoxic rats. Experimental design as follows: Group I, saline control for 21 days; Group II, gentamicin nephrotoxic control for eight days; Group III, alcoholic extract of W. somnifera for 13 days + simultaneous administration of gentamicin and W. somnifera, from day 14 to 21 (nephroprotective) and Group IV, gentamicin for 8 days + alcoholic extract of W. somnifera from day 9 to 21 (nephrocurative). End of experiment, respective serum and kidney tissue samples used to analyze renal function. Withania somnifera as a nephroprotective and nephrocurative molecule significantly restore the renal function on gentamicin-induced nephrotoxicity. This phenomenon is accompanied with significantly reduced blood urea nitrogen, creatine, alkaline phosphatase, gamma-glutamyl transferase, albumin, total protein, calcium, potassium and kidney malondialdehyde concentrations. Additionally, W. somnifera significantly increased antioxidant activities of glutathione and superoxide dismutase to protect renal tissue damage from gentamicin in wistar rats. Over all, W. somnifera treated nephroprotective animal shows improved recovery compared to nephrocuartive. The nephroprotective or nephrocurative effect of W. somnifera could be due to inherent antioxidant and free-radical-scavenging principle(s). In the near future, biologically active compounds of W. somnifera (withanolides) could appear as a novel therapeutic molecule for renal disorders.

Resumo em Inglês:

ABSTRACT Hyperuricemia is the main cause of gout, an inflammation induced by uric acid deposition in joints. Drugs available present side effects, so there is a need for new treatment alternatives. Lychnophora species are used in folk medicine to treat inflammation, rheumatism and muscle pain. Goyazensolide (10 mg/kg), eremantholide C (25 mg/kg) and lychnopholide (25 mg/kg), sesquiterpene lactones isolated from Lychnophora species were previously studied and showed anti-hyperuricemic effects in mice. However, the mechanisms of this effect were not elucidated. The methodology of this study consisted in treatment of hyperuricemic-induced rats, and comparison between control groups, clinically used anti-hyperuricemic drugs and sesquiterpene lactones. Urine and blood were collected for uric acid quantification. Xanthine oxidase inhibition was measured in liver homogenates. Results showed that all evaluated sesquiterpene lactones presented anti-hyperuricemic activity at the doses of 5 and 10 mg/kg and can act through one or both mechanisms, depending on the dose administrated. Goyazensolide and lychnopholide at dose of 5 mg/kg showed important uricosuric effect. Goyazensolide and lychnopholide at dose of 10 mg/kg, and eremantholide C (5 and 10 mg/kg) presented notable inhibition of hepatic xanthine oxidase activity and uricosuric effect. Thus, these sesquiterpene lactones are promising hypouricemic agent to treat hyperuricemia and gout.

Resumo em Inglês:

ABSTRACT Smilax moranensis M.Martens & Galeotti, Smilacaceae, root is a medicinal plant used among the Chatinos in Oaxaca, Mexico, to control type 2 diabetes. The objectives of the study were to isolate the bioactive compounds from the roots of Smilax and evaluate the chronic hypoglycemic effect of the ethanol–water extract. The main compounds were isolated from the methanolic extract via conventional phytochemical methods. The dried roots of S. moranensis were extracted with methanol and chromatographed on Sephadex LH 20. Fractions were chromatographed and purified on a silica gel chromatography column. The ethanol–water extract was orally administered to hyperglycemic rats for a period of 42 days, and glucose, glycated hemoglobin (HbA1c), and triacylglycerides were measured. Moreover, very-low-density lipoprotein was calculated. During the chemical investigation, three compounds were isolated and characterized, namely, 3-O-caffeoyl-quinic acid, 5-O-caffeoyl-quinic acid and trans-resveratrol, using various spectroscopic techniques. Animal experiments confirmed that the plant extract could control both the glucose and HbA1c levels. In conclusion, this study confirms that the roots of S. moranensis have hypoglycemic properties and suggests that the isolated compounds are potentially involved in this effect.

Resumo em Inglês:

ABSTRACT Centella asiatica (L.) Urb., Apiaceae, is commonly used as food, food supplement, and medicine. Development of the extraction process to obtain the high extent of the active compound is necessary. So, the response surface methodology was used in this work to optimize the dynamic maceration of C. asiatica to obtain the highest content of the four centelloids including asiatic acid, madecassic acid, asiaticoside, and madecassoside. Two factors: extraction temperature and extraction time, were studied. The content of four centelloids was observed. After the extraction of C. asiatica using ethanol, the content of four centelloids was analyzed using validated high performance liquid chromatography. The optimization result showed that madecassoside and asiaticoside had a similar pattern of the contour plots and response surfaces. These two centelloids were highly extracted at a high extraction time and high extraction temperature. The other two centelloids had the same pattern, they had a high content at a high temperature and time as well as at a low temperature and time. The simultaneous highest content of four centelloids was achieved when extracted at 60 °C for 120 min. The optimal condition could be used as standard condition for extraction of C. asiatica to provide the highest content of four centelloids.

Resumo em Inglês:

ABSTRACT Hippeastrum goianum (Ravenna) Meerow, Amaryllidaceae, is an endemic species from the Cerrado, Brazil; there are only few studies about its chemistry or biological activity. This study aimed to investigate the occurrence of lycorine in extracts from in vitro H. goianum plantlets, as well as evaluate a possible inhibition of acetylcholinesterase. The ethanol extract of plantlets produced by in vitro seed germination and micropropagation of bulblets was obtained from seedlings from in vitro germination, while the ethanol extract micropropagtion of bulblets was obtained from a subculture of those seedlings. The presence of lycorine was detected in only in the micropropagation of bulblets. The micropropagation of bulblets was more active than the plantlets produced by in vitro seed germination, with an IC50 of 114.8 ± 0.95 µg/ml and IC50 386.00 ± 0.97 µg/ml, respectively. These results showed that both in vitro germination and micropropagation of H. goianum can lead to the biosynthesis of lycorine. Moreover, the micropropagation led to improved anticholinesterase activity.

Resumo em Inglês:

ABSTRACT Dysphania ambrosioides (L.) Mosyakin and Clemants (Syn: Chenopodium ambrosioides L.), Amaranthaceae, is a plant with antibacterial, antifungal, antioxidant, antiparasitic and antitumor properties that is commonly used in Brazilian folk medicine. In this work we performed the optimization of ultrasound-assisted extraction of flavonoids in the aerial parts of D. ambrosioides. The flavonoid concentrations, as rutin equivalents, were quantified with the aid of a validated spectrophotometric method. The Box–Behnken (33) design with response surface methodology, for the independent variables, extraction time, temperature, and ethanol content, were used for the optimization of ultrasound-assisted extraction. The analytical method was selective, linear, without matrix interference, accurate, precise and robust. The best conditions for the ultrasound-assisted extraction of flavonoids were: time of 60 min, temperature of 57 °C and ethanol content of 57% (w/w). The methods of extracting and quantifying flavonoids developed in the present study have provided be eco-friendly, simple, and useful to determine the flavonoid content, expressed as rutin equivalents, in the aerial parts of D. ambrosioides.