Use of HPLC–DAD–ESI/MS to profile phenolic compounds in edible wild greens from Portugal

In some Mediterranean areas traditional wild greens are responsible for a significant percentage of total dietary antioxidant intakes. Asparagus acutifolius L. (wild asparagus), Bryonia dioica Jacq. (white bryony) and Tamus communis L. (black bryony) are important examples of those edible wild greens widely consumed. This study aimed to determine the phenolic profile and composition of edible vernal early shoots of those species. Wild asparagus and black bryony revealed glycosides of flavonols as the main phenolic compounds, while white bryony contained C-glycosylated flavones. Black bryony was the wild green that possessed the highest content of phenolic compounds (2200 mg/kg). Amongst the 11 flavonols found in this sample, kaempferol glycosides were the main compounds (1760 mg/kg). In the sample of wild asparagus, quercetin 3-O-rutinoside was the main flavonol found (263 mg/kg). Five flavones and one flavonol were found in the white bryony sample, apigenin 6-C-glucoside-7-O-glucoside being the major compound (1550 mg/kg).     

Flavonol glycosides from whole cottonseed by-product

Cottonseeds are fed to high-producing dairy cows as a source of fat and highly-digestible fibre. Seven flavonol glycosides have been identified from whole cottonseed by-product. Their structures were established as quercetin 3-O-{β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside} (1), kaempferol 3-O-{β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside} (2), quercetin 3-O-[β-d-apiofuranosyl-(1 → 2)-β-d-glucopyranoside] (3), quercetin 3-O-β-d-glucopyranoside (4), kaempferol 3-O-[α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranoside] (5), quercetin 3-O-[α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranoside] (6), and kaempferol 3-O-α-l-rhamnopyranoside (7). Gallic acid (8) and 3,4-dihydroxybenzoic acid (9) were also found. All structures were elucidated by ESI-MS and NMR spectroscopic methods. Total polyphenols were assayed by the Folin–Ciocalteu method.     

α-Glucosidase inhibitors from Devil tree (Alstonia scholaris)

α-Glucosidase inhibitors are used in the treatment of non-insulin-dependent diabetes mellitus. We attempt to isolate α-glucosidase inhibitors from 24 traditional Thai medicinal plant samples. Potent α-glucosidase inhibitory activity was found in aqueous methanol extract of dried Devil tree (Alstonia scholaris) leaves. Active principles against α-glucosidase, prepared from rat small intestine acetone powder, were isolated and identified. The structures of these isolated compounds were found to be quercetin 3-O-β-d-xylopyranosyl (1? → 2″)-β-d-galactopyranoside and (−)-lyoniresinol 3-O-β-d-glucopyranoside on the basis of chemical and spectral evidence. The latter exhibited an inhibitory activity against both sucrase and maltase with IC₅₀ values of 1.95 and 1.43 mM, respectively, whereas the former inhibited only maltase with IC₅₀ values of 1.96 mM. This preliminary observation will provide the basis for further examination of the suitability of Alstonia scholaris as a medicinal supplement that contributes toward the treatment and prevention of diabetes.     

Chemical properties of the cultivated Sideritis raeseri Boiss. & Heldr. subsp. raeseri

Phytochemical analyses of the cultivated Sideritis raeseri subsp. raeseri in four different stages of flower development were performed. Traditionally used infusion and decoction were also prepared from aerial parts in full flowering stage, and analyses of active compounds and radical scavenging capacity were performed. The highest yield of the essential oil, obtained by hydrodistillation, was noticed in the full flowering phase (0.11%), with sesquiterpene bicyclogermacrene as the main constituent (42.5%). All examined extracts contained phenolic compounds and their amounts varied from 15.3 to 34.1 mg GAE/g DW. The amounts of total phenolics in infusion and decoction were similar (46.5 and 43.9 mg GAE/100 ml, respectively). LC–ESI-MS analyses of all samples allowed the characterisation of 22 phenolic compounds. Two dominant flavone glycosides, 4′-O-methylhypolaetin-7-O-[6?-O-acetyl-β-d-allopyranosyl (1 → 2)-β-d-glucopyranoside (17) and 4′-O-methylisoscutellarein-7-O-[6?-O-acetyl-β-d-allopyranosyl-(1 → 2)]-β-d-glucopyranoside (19) were quantified using HPLC. Moreover, the mineral content and the percent of transportation were investigated.     

Identification,quantification and antioxidant activity of acylated flavonol glycosides from sea buckthorn (Hippophae rhamnoides ssp. sinensis)

A novel acylated flavonol glycoside: isorhamnetin (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (1), together with two known acylated flavonol glycosides: quercetin (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (2) and kaempferol (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (3) were isolated from the n-butanol fraction of sea buckthorn (Hippophae rhamnoides ssp. sinensis) berries for the first time by chromatographic methods, and their structures were elucidated using UV, MS, ¹H and ¹³C NMR, and 2D NMR. Compounds 1–3 showed good scavenging activities, with respective IC₅₀ values of 8.91, 4.26 and 30.90 μM toward the 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radical; respective Trolox equivalent antioxidant capacities of 2.89, 4.04 and 2.44 μM μM⁻¹ toward 2,2′-azino-bis-3-ethyl-benzothiazoline-6-sulphonate (ABTS) radical. The quantitative analysis of the isolated acylated flavonol glycosides was performed by HPLC–DAD method. The contents of compounds 1–3 were in the range of 12.2–31.4, 4.0–25.3, 7.5–59.7 mg/100 g dried berries and 9.1–34.5, 75.1–182.1, 29.2–113.4 mg/100 g dried leaves, respectively.     

Anti-complementary activity of flavonoids from Gnaphalium affine D. Don

Gnaphalium affine D. Don, is used as a vegetable as well as a folk medicine in China for its anti-inflammatory, antitussive, and expectorant activities. Phytochemical investigation of the ethyl acetate (EtOAc) fraction of this plant led to isolation of three new acylated flavonol glycosides, apigenin 4′-O-β-d-(6″-E-caffeoyl)-glucopyranoside, luteolin 4′-O-β-d-(6″-E-caffeoyl)-glucopyranoside, and quercetin 4′-O-β-d-(6″-E-caffeoyl)-glucopyranoside, together with 24 known compounds. The structures of these compounds were determined by spectroscopic methods. All compounds were evaluated for their anti-complementary activity on the classical pathway of the complement system in vitro, and luteolin 4′-O-β-d-(6″-E-caffeoyl)-glucopyranoside exhibited highest anti-complementary activity with an IC₅₀ value of 0.045 ± 0.005 mg/ml. This is the first report of anti-complementary activity of G. affine D. Don, which displays the initial evidence that this plant is a potent complement inhibitor.     

A novel mechanism for the promotion of quercetin glycoside absorption by megalo α-1,6-glucosaccharide in the rat small intestine

The presence of an α-1,6-glucosaccharide enhances absorption of water-soluble quercetin glycosides, a mixture of quercetin-3-O-β-d-glucoside (Q3G, 31.8%), mono (23.3%), di (20.3%) and more d-glucose adducts with α-1,4-linkage to a d-glucose moiety of Q3G, in a ligated small intestinal loop of anesthetized rats. We prepared α-1,6-glucosaccharides with different degrees of polymerization (DP) enzymatically and separated them into a megalo-isomaltosaccharide-containing fraction (M-IM, average DP = 11.0) and an oligo-isomaltosaccharide-containing fraction (O-IM, average DP = 3.6). Luminal injection of either saccharide fraction promoted the absorption of total quercetin-derivatives from the small intestinal segment and this effect was greater for M-IM than O-IM addition. M-IM also increased Q3G, but not the quercetin aglycone, concentration in the water-phase of the luminal contents more strongly than O-IM. The enhancement of Q3G solubilization in the luminal contents may be responsible for the increases in the quercetin glucoside absorption promoted by α-1,6-glucosaccharides, especially that by M-IM. These results suggest that the ingestion of α-1,6-glucosaccharides promotes Q3G bioavailability.     

Antihypertensive properties of flavonoid-rich apple peel extract

Hypertension is a major public health problem rising across the globe. Inhibition of angiotensin converting enzyme (ACE) is identified as a main therapeutic target in controlling high blood pressure. The present study investigated the ACE inhibitory property of a flavonoid-rich apple peel extract (FAE), its constituents, selected flavonoids and some quercetin metabolites using a biochemical assay of ACE inhibition and a human umbilical vein endothelial cell (HUVEC) model. FAE, all the tested flavonoids except genistein, and two quercetin metabolites (quercetin-3-O-glucuronic acid and quercetin-3-O-sulfate) significantly (p < 0.05) inhibited ACE. Enzyme kinetic analysis revealed that flavonoids are competitive inhibitors of ACE. In the HUVEC model, FAE, quercetin-3-O-glucoside and quercetin-3-O-glucuronic acid inhibited significantly (p < 0.05) ACE activity. Overall, FAE and most of the flavonoids tested showed ACE inhibition in vitro which needs further investigations using animal and human clinical trials.     

Antithrombotic activity of fractions and components obtained from raspberry leaves (Rubus chingii)

The 70% ethanol fraction from an aqueous extract of raspberry leaves was shown to be the most antithrombotic fraction in in vitro and in vivo tests. The total flavonoids and phenolics in this fraction were 0.286 g/g and 0.518 g/g by colorimetry. Six compounds, including salicylic acid, kaempferol, quercetin, tiliroside, quercetin 3-O-β-d-glucopyranoside and kaempferol 3-O-β-d-glucopyranoside, were isolated from the active fraction. Among them, kaempferol, quercetin and tiliroside obviously delayed plasma recalcification time (PRT) in blood.     

Characterization of phenolic compounds in Chinese hawthorn (Crataegus pinnatifida Bge. var. major) fruit by high performance liquid chromatography–electrospray ionization mass spectrometry

Hawthorn (Crataegus spp.) fruits are increasingly popular as raw materials for nutraceuticals and functional foods. As a major group of bio-active components of hawthorn, the phenolic compounds of the fruits have not been well characterized so far. After extraction with 80% aqueous ethanol, the phenolics of the fruits of a major Chinese hawthorn variety, Crataegus pinnatifida Bge. var. major, were separated by polyamide column chromatography, followed by analyses by high performance liquid chromatography combined with diode array UV spectrometry and electrospray ionization mass spectrometry. Ideain (cyanidin-3-O-galactoside), chlorogenic acid, procyanidin B2 [epicatechin-(4β → 8)-epicatechin], epicatechin, hyperoside (quercetin-3-O-galactoside) and isoquercitrin (quercetin-3-O-glucoside) were identified with UV spectra, mass spectra and reference compounds. In addition, 35 compounds were tentatively identified based on UV and mass spectra. These compounds were mostly B-type procyanidins (PA) and their glycosides including aglycons of 3 dimers, 3 trimers, 8 tetramers, 4 pentamers, 2 hexamers and 2 glycosides of PA monomers, 7 glycosides of PA dimers, 1 glycoside of a PA trimer, 2 glycosides of PA tetramers, 1 glycoside of a PA pentamer, and 2 glycosides of quercetin. This is the first systematic study of phenolic compounds in Chinese hawthorn fruits and the first report of the presence of glycosylated procyanidins in hawthorn.     

The effect of temperature and radiation on flavonol aglycones and flavonol glycosides of kale (Brassica oleracea var. sabellica)

The winter crop kale has a complex profile of different glycosylated and acylated flavonol glycosides which may be affected by global warming. To the best of our knowledge, compound–climate relationships for flavonol aglycones and flavonol glycosides were established for the first time. The investigated 10 major flavonol glycosides responded structure-dependent in the investigated temperature range between 0 and 12 °C and the photosynthetic active radiation range between 4 and 20 mol m⁻² d⁻¹, e.g. the decrease in temperature led to an increase in sinapic acid monoacylated and diacylated quercetin glycosides, while the sinapic acid monoacylated kaempferol glycosides showed a maximum at 4.5 °C. Furthermore, the hydroxycinnamic acid residues and the different number of glucose moieties in the 7-O position affected the response of kaempferol triglucosides. Consequently, global warming would result in lower concentrations of antioxidant-relevant quercetin glycosides in winter crops, suggesting a production at e.g. higher altitudes due to lower temperature.     

HPLC-DAD-ESIMS analysis of phenolic compounds in bayberries (Myrica rubra Sieb. et Zucc.)

Qualitative analysis of the ethyl acetate extracts from three bayberry cultivars, Xiangshan, Biqi and Dongkui, was performed by means of a hyphenated technique of HPLC coupled to photodiode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESIMS). Three phenolic compounds were identified (gallic acid, protocatechuic acid, and quercetin 3-glucoside) and seven others (two myricetin hexoside and two myricetin deoxyhexoside derivatives; quercetin hexoside and quercetin deoxyhexoside derivatives; kaempferol hexoside derivative) partially identified. Quantification of phenolic compounds was performed by HPLC-DAD, which revealed that gallic acid (2.6–7.0 mg/kg FW) was the major phenolic acid in all analysed cultivars. Myricetin glycosides (71.1 mg/kg FW) were the major flavonol glycosides in cultivar Xiangshan and quercetin glycosides (117.8 mg/kg FW) were the major ones in cultivar Biqi. Cultivar Dongkui had medium contents of quercetin glycosides (48.0 mg/kg FW) and myricetin glycosides (53.2 mg/kg FW). Kaempferol glyosides (3.1–4.6 mg/kg FW) were the lowest contents of flavonol glycosides in the assayed bayberries. These results are relevant not only from a nutritional point of view, but also in the control of color stability and haze formation during bayberry juice processing and storage.     

Flavonoid glycosides from Pouteria obovata (R. Br.) fruit flour

Three unknown dihydroflavanol glycosides: 2R,3R-4′O-methyl dihydrokaempferol 7-O-[3″-O-acetyl]-β-d-glucopyranoside (1), 2R,3R-4′-O-methyl dihydrokaempferol 7-O-β-d-β-l-xylopyranosyl-(1″′ → 6″)-[3″-O-acetyl]-β-d-glucopyranoside (2), 2R,3R-4′-O-methyl dihydrokaempferol 3-O-β-d-β-l-xylopyranosyl-(1″′ → 6″)-[3″-O-acetyl]-β-d-glucopyranoside (3), together with gallic acid (4) were isolated from the n-butanol fraction of Pouteria obovata fruit flour by chromatographic methods and their structures were elucidated on the basis of CD, UV, MS, monodimensional NMR (¹H and ¹³C) and bidimensional NMR (COSY, HSQC and HMBC). The quantitative analysis of flavonoids and phenols were also reported. Total phenolic amount (51.1 ± 14.1 mg GAE/1000 g; p < 0.0006) and flavonoid content (153.2 ± 3.5 mg CE/100 g; p < 0.004) were detected spectrophotometrically.     

Antioxidant and cytotoxic flavonoids from the flowers of Melastoma malabathricum L.

Phytochemical and bioactivity studies of the flowers of Melastoma malabathricum L. (Melastomataceae) have been carried out. The ethyl acetate extract yielded three compounds, identified as naringenin, kaempferol and kaempferol-3-O-d-glucoside, and methanol extract gave kaempferol-3-O-(2″,6″-di-O-p-trans-coumaroyl)glucoside and kaempferol-3-O-d-glucoside. The crude extracts and isolated compounds were screened for their antioxidant and cytotoxic activities. The antioxidant assay was carried out by the DPPH radical-scavenging electron spin resonance (ESR) spectroscopic method. The cytotoxicity was measured by the MTT assay against a MCF7 cell line. Naringenin, kaempferol, kaempferol-3-O-d-glucoside, kaempferol-3-O-(2″,6″-di-O-p-trans-coumaroyl) glucoside, ethyl acetate and methanol extracts were found to be active as radical-scavengers with IC₅₀ values of 0.52 mM, 81.5 μM, 1.07 mM, 35.8 μM, 7.21 μg/ml and 6.59 μg/ml, respectively. Naringenin and kaempferol-3-O-(2″,6″-di-O-p-trans-coumaroyl)glucoside were also found to be active in inhibiting cell proliferation of MCF7 with IC₅₀ values of 0.28 μM and 1.3 μM, respectively.     

Antioxidative properties and flavonoid composition of Chenopodium quinoa seeds cultivated in Japan

To evaluate the nutritional advantages of quinoa seeds (Chenopodium quinoa Willd.) cultivated in Japan, antioxidative properties and flavonoid composition were determined and compared to corresponding data for conventionally-used cereals and pseudo-cereals, including quinoa seeds from South America. The antioxidant activities of these grains against DPPH radicals were strongly associated with the total phenolic content of the tested samples. The crude extracts of quinoa seeds cultivated in Japan exhibited higher antioxidative effects than those from South America and other cereals, excluding buckwheat. Four flavonol glycosides were isolated and identified from the Japanese quinoa seeds, and the chemical composition of the flavonoids – quercetin and kaempferol 3-O-(2″,6″-di-O-α-rhamnopyranosyl)-β-galactopyranosides (1 and 4), quercetin 3-O-(2″,6″-di-O-α-rhamnopyranosyl)-β-glucopyranoside (2), and quercetin 3-O-(2″-O-β-apiofuranosyl-6″-O-α-rhamnopyranosyl)-β-galactopyranoside (3) – was evaluated through quantitative determination. Trioside 2 was isolated for the first time from quinoa seeds. These glycosides were not detected in extracts from any of the tested grains except quinoa. The aglycone quercetin content of the Japanese quinoa seeds is higher than in the seeds from South America and buckwheat. The amounts of quercetin and kaempferol formed via acidic hydrolysis in quinoa are much higher than those of conventionally-used edible plants. The quinoa seeds cultivated in Japan are the most effective functional foodstuff – in terms of being a source of antioxidative and bioactive flavonoids – among cereals and pseudo-cereals.     

Identification of phenolic compounds isolated from pigmented rices and their aldose reductase inhibitory activities

Two anthocyanins (cyanidin-3-O-β-glucoside and peonidin-3-O-β-glucoside) and other phenolic (ferulic acid) were, respectively isolated from black and pigmented brown rices (Oryza sativa L. japonica) and their complete structures were determined by spectroscopic analyses (H NMR, C NMR and MALDI MASS). The HPLC profile of anthocyanins extracted from black rice showed cyanidin-3-O-β-glucoside as the first peak (85%) and peonidin 3-O-β-d-glucoside as the second (15%), while that of pigmented brown rice showed ferulic acid as the first peak (85.7%) and tocols as the second (14.3%). Several tocols were isolated and identified from the unsaponifiable fractions of both rices having some difference on their structures and amounts. The aldose reductase inhibitory activity of isolated compounds was in the following decreasing order: cyanidin-3-glucoside > quercetin > ferulic acid > peonidin-3-glucoside > tocopherol.     

Spirostane and cholestane glycosides from the bulbs of Allium nigrum L

A phytochemical investigation of the fresh bulbs of Allium nigrum L. led to the isolation of new spirostane-type glycosides as two inseparable isomer mixtures, nigrosides A1/A2 (1a/1b) and nigrosides B1/B2 (2a/2b), two new cholestane-type glycosides, nigrosides C and D (3 and 4), together with the known compounds, 25(R,S)-5α-spirostan-2α,3β,6β-trio1-3-O-β-d-glucopyranosyl-(1 → 2)-O-[β-d-xylopyranosyl-(1 → 3)]-O-β-d-glucopyranosyl-(1 → 4)-β-d-galactopyranoside (5a/5b) and 25(R,S)-5α-spirostan-2α,3β,6β-trio1 3-O-β-d-glucopyranosyl-(1 → 2)-O-[4-O-(3S)-3-hydroxy-3-methylglutaryl-β-d-xylopyranosyl-(1 → 3)]-O-β-d-glucopyranosyl-(1 → 4)-β-d-galactopyranoside (6a/6b), isolated from this plant for the first time. All structures were elucidated mainly by spectroscopic analysis (1D and 2D NMR experiments, FABMS, HRESIMS) and by comparison with literature data. Cytotoxicity of the isolated compounds was assessed against human colon carcinoma (HT-29 and HCT 116) cell lines. Compounds 5a/5b and 6a/6b were found to be the most active with IC₅₀ values 1.09 and 2.82 μM against HT-29 and 1.59 and 3.45 μM against HCT 116, respectively.     

Anti-melanogenesis properties of quercetin- and its derivative-rich extract from Allium cepa

In an effort to find a new whitening agent, we have found that the methanol extract of the dried skin of Allium cepa showed inhibition of melanin formation. Bioassay-guided fractionation led to the isolation of quercetin (1) and quercetin 4’-O-β-glucoside (3) from A. cepa as the inhibitors of melanin formation in B16 melanoma cells with IC₅₀ values of 26.5 and 131 μM, respectively. In addition, we evaluated the effect of some quercetin derivatives, such as isoquercitrin (2), quercetin 3,4’-O-diglucoside (4), rutin (5) and hyperin (6) on B16 melanoma cells. These quercetin derivatives did not show any inhibition of melanin formation. Furthermore, the ORAC values of compounds 1–6 were 7.64, 8.65, 4.82, 4.32, 8.17 and 9.34 μmol trolox equivalents/μmol, respectively. Dried skin of red onion showed inhibitory activity against melanin formation in B16 melanoma cells, as well as antioxidant properties.     

The antioxidant and free-radical scavenging activities of extract and fractions from corn silk (Zea mays L.) and related flavone glycosides

This study was designed to evaluate both the antioxidant and free-radical scavenging activities of extract and fractions from corn silk. N-butanol fraction (BF) demonstrated the highest total phenolic content (164.1 ± 9.7 μg GAE/g DCS) and total flavonoids content (69.4 ± 5.1 μg RE/g DCS), accompanied with the highest antioxidant activity compared to other fractions through all antioxidant assays. Two flavone glycosides showing potent antioxidant activity were isolated from BF and identified, by comparing spectral data (UV, FAB-MS and NMR) with literature values, to be isoorientin-2″-O-α-l-rhamnoside and 3′-methoxymaysin. The two isolated flavone glycosides, particularly isoorientin-2″-O-α-l-rhamnoside, demonstrated significant total antioxidant activity, DPPH radical scavenging activity, reducing power and iron-chelating capacity, with EC₅₀ values of 14.24 ± 1.49, 22.69 ± 2.33, 6.58 ± 1.07 and 30.25 ± 3.05 μg/ml, respectively. Results obtained indicated that corn silk extracts can be used potentially as a ready accessible and valuable bioactive source of natural antioxidants.     

Phytochemical profiles and inhibitory effect on free radical-induced human erythrocyte damage of Dracaena draco leaf: A potential novel antioxidant agent

The present study reports for the first time the metabolite profile and antioxidant activity of aqueous extract obtained from Dracaena draco L. leaf. Volatiles profile was determined by HS-SPME/GC-IT-MS, with 34 compounds being identified, distributed by distinct chemical classes: 2 alcohols, 5 aldehydes, 16 carotenoid derivatives and 8 terpenic compounds. Carotenoid derivative compounds constituted the most abundant class in leaf (representing 45% of total identified compounds). Phenolics profile was determined by HPLC/DAD and 9 constituents were identified: 2 hydroxycinnamic acid derivatives – 5-O-caffeoylquinic and 3,5-O-dicaffeoylquinic acids; 4 hydroxycinnamic acids – caffeic, p-coumaric, ferulic and sinapic acids and 3 flavonol glycosides – quercetin-3-O-rutinoside, kaempferol-3-O-glucoside and kaempferol-3-O-rutinoside. The most abundant phenolic compound is quercetin-3-O-rutinoside (representing 50.2% of total polyphenols). Organic acids composition was also characterised, by HPLC–UV and oxalic, citric, malic and fumaric acids were determined. Oxalic and citric acids were present in higher amounts (representing 47%, each). The antioxidant potential of this material was assessed by the ability to protect against free radical-induced biomembrane damage, using human erythrocyte as in vitro model. Leaf extract strongly protected the erythrocyte membrane from haemolysis (IC₅₀ of 39 ± 11 μg/ml), in a time- and concentration-dependent manner. This is the first report showing that D. draco leaf is a promising antioxidant agent.