In vitro determination of antioxidant activity of proteins from jellyfish Rhopilema esculentum

In this study, the antioxidant activity of proteins isolated from jellyfish, Rhopilema esculentum Kishinouye (R. esculentum), was determined by various antioxidant assays, including superoxide anion radical-scavenging, hydroxyl radical-scavenging, total antioxidant activity, reducing power and metal chelating activity. Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol, vitamin C and mannitol were used as standards in those various antioxidant activities. The crude protein (CP) and the protein fractions isolated by Sephadex chromatography, first peak (FP) and second peak (SP), had very low reductive power and metal chelating abilities compared to EDTA, but they showed strong scavenging effects on the superoxide anion radical, hydroxyl radical and varying total antioxidant activity. FP and SP exhibited stronger scavenging effects on the superoxide anion radical than BHA, BHT or α-tocopherol. The EC₅₀ values of FP and SP were 6.12 and 0.88 μg/ml, respectively, while values EC₅₀ of BHA, BHT and α-tocopherol were 31, 61 and 88 μg/ml, respectively. CP, FP and SP showed far higher hydroxyl radical-scavenging activities than did vitamin C or mannitol. The EC₅₀ values of CP, FP and SP were 48.76, 45.42 and 1.52 μg/ml, but EC₅₀ values of vitamin C and mannitol were 1907 and 4536 μg/ml, respectively. In a β-carotene–linoleate system, SP and CP showed antioxidant activity, but lower than BHA. Of the three samples, SP had the strongest antioxidant activity. So, SP may have a use as a possible supplement in the food and pharmaceutical industries.     

Microwave-assisted ionic liquids pretreatment followed by hydro-distillation for the efficient extraction of essential oil from Dryopteris fragrans and evaluation of its antioxidant efficacy in sunflower oil storage

With strong dissolution abilities of ionic liquids to plant cell walls, an integrated microwave-assisted ionic liquids pretreatment followed by hydro-distillation method (MILP–MHD) was performed for the efficient extraction of essential oil from the rare Dryopteris fragrans. Under optimal conditions, this innovative MILP–MHD provided the remarkable superiority of much higher yield (0.91%) of D. fragrans essential oil (DEO) within much shorter extraction duration (14.2 min) than those of the reported solvent-free microwave extraction technique (0.33% and 94 min). Furthermore, the antioxidant efficacy of DEO in the stabilization of sunflower oil was determined and found to be comparable to the commonly-employed α-tocopherol (VE) but a little lower than the synthetic antioxidant butylated hydroxytoluene (BHT). Therefore, MILP–MHD is a potential and highly efficient technique for the extraction of valuable essential oils in flavor food industries and DEO can be explored as a potent natural antioxidant additive for edible oils or lipid foods storage.     

Antimicrobial and antioxidant properties of the essential oils and methanol extract from Mentha longifolia L. ssp. longifolia

This study was designed to evaluate antimicrobial and antioxidant activities of the essential oil and methanol extract from Mentha longifolia ssp. longifolia. The essential oil showed strong antimicrobial activity against all 30 microorganisms tested whereas the methanol extract almost remained inactive. In contrast, the extract showed much better activity than the essential oil in antioxidant activity assays employed, e.g. in the inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene/linoleic acid systems. In the former, the extract was able to reduce the stable free radical DPPH with an IC₅₀ of 57.4 μg/ml while that of the oils was 10 700 μg/ml. When compared to BHT, a synthetic antioxidant, both showed weaker antioxidative potential. Similarly, in β-carotene/linoleic acid assay, these samples were not effectively able to inhibit the linoleic acid oxidation; exhibiting only 24% and 36% inhibitions at 2 mg/ml, respectively; both were far below than that of BHT. Total phenolic constituent of the extract was 4.5 g/100 g as gallic acid equivalent. GC–MS analysis of the oil resulted in the identification of 45 constituents, cis-piperitone epoxide, pulegone and piperitenone oxide being the main components.     

Evaluation of the antioxidant activity of three microalgal species for use as dietary supplements and in the preservation of foods

The antioxidant activity of the microalgal ethanolic extracts of Porphyridium cruentum, Phaeodactylum tricornutum and Chlorella vulgaris was determined by means of the β-carotene–linoleate model system. The results show that the activity of C. vulgaris extract was higher than those obtained for the other microalgal extracts tested and for the synthetics BHA (butylated hydroxyanisole), and BHT (butylated hydroxytoluene). In addition, the major constituents present in the ethanolic extracts of the three microalgae species were analyzed by means of GC and GC–mass spectrometry. The results showed that the tested microalgae may be an important source of natural antioxidants, as an alternative to higher plants or the production by chemical synthesis.     

The in vitro antioxidative properties of the essential oils and methanol extracts of Satureja spicigera (K. Koch.) Boiss. and Satureja cuneifolia ten

This study was designed to examine the in vitro antioxidant activities of the essential oil and methanol extracts of Satureja spicigera and S. cuneifolia from Turkish flora. GC and GC/MS analysis of the essential oils resulted in the identification of 40 and 29 compounds, representing the 99.4% and 99.5% of the oils, respectively. Major constituents of the oils were carvacrol (42.5% and 67.1%), γ-terpinene (21.5% and 15.2%) and p-cymene (20.9% and 6.7%), respectively. Methanol extracts were also obtained from the aerial parts of the plants. The samples were subjected to a screening for their possible antioxidant activities by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene–linoleic acid assays. In general, samples obtained from S. cuneifolia exerted greater antioxidant activities than did those obtained from S. spicigera. In the DPPH test system, free radical-scavenging activity of S. spicigera oil was determined to be 127 ± 1.63 μg/ml, whereas IC₅₀ value of S. cuneifolia was 89.1 ± 2.29 μg/ml. In the β-carotene–linoleic acid test system, antioxidant activities of the oil were 81.7 ± 1.14% and 93.7 ± 1.83%, respectively. Antioxidant activities of the synthetic antioxidant, BHT, ascorbic acid, curcumin and α-tocopherol were also determined in parallel experiments.     

GC/MS analysis and in vitro antioxidant activity of essential oil and methanol extracts of Thymus caramanicus Jalas and its main constituent carvacrol

Chemical composition of the essential oil, antioxidant activity (DPPH and β-carotene/linoleic acid assays), and total phenolic content (Folin–Ciocalteu assay) of aerial parts of Thymus caramanicus were determined. The highest radical-scavenging activity (DPPH test) was shown by the polar subfraction of the methanol extract (IC₅₀ = 43.0 μg/ml) which was also higher than that of butylated hydroxytoluene (BHT, IC₅₀ = 19.7 μg/ml). However, it was the nonpolar subfraction of the methanol extract that showed the highest inhibition (84.4%), as assessed by the β-carotene/linoleic acid assay, which was only slightly lower than that shown by BHT (93.3%). The antioxidant activities of the essential oil main component (carvacrol) were also evaluated for comparison. Total phenolic content of the polar subfraction, as gallic acid equivalents, was 124.3 μg/mg. Essential oil extracted from the aerial parts by hydrodistillation was analysed by GC and GC/MS. Fifteen constituents, representing 99.3% of the oil, were identified, of which the major ones, carvacrol (85.9%), thymol (3.3%), p-cymene (3.2%), γ-terpinene (1.8%) and borneol (1.3%), accounted for 95.6% of the oil.     

Determination of antioxidative potential of lichen Usnea ghattensis in vitro

The study was aimed at evaluating the antioxidant activities of extract of Usnea ghattensis. The antioxidant activity, reducing power, superoxide anion radical scavenging and free radical scavenging activities were studied. The antioxidant activity increased with the increasing amount of extracts (2-20 mg/ml) added to the linoleic acid emulsion. Lipid peroxidation upto 73.3% was inhibited by the extract of 20 mg/ml and 39.2% by α-tocopherol at the same concentration. These effects were statistically significant (r²=0.876,P<0.01) when compared with control. However, the extract had no significant effect on superoxide anion scavenging by the PMS/NADH-NBT method. Like antioxidant activity, the reducing power and free radical scavenging activity of extract depends on concentration and increasing with increased amount of sample. The reducing power and DPPH radical-scavenging activity of U. ghattensis extract were found greater than the BHA and BHT. The results obtained in the present study indicate that U. ghattensis is a potential source of natural antioxidant.     

Antioxidant activity of ethanolic extract of Cortex fraxini and use in peanut oil

Cortex fraxini was extracted with 95% ethanol to obtain a crude antioxidant extract. The antioxidant activity was evaluated using the linoleic acid peroxidation method and the free radical scavenging assays, namely 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. Cortex fraxini extract (CFE) showed high inhibition of peroxidation of linoleic acid when compared with butylated hydroxytoluene (BHT). CFE also exhibited excellent scavenging activity on DPPH and hydroxyl radicals. Total antioxidant activity was measured by the reduction of Mo(VI) to Mo(V) by the extract, and subsequent formation of a green phosphate/Mo(V) complex at acid pH. CFE had significant total antioxidant activity and the effects were increased with increasing reaction time. The total phenolic content of the sample, analyzed by using Folin–Ciocalteu’s reagent, was 91.33 mg/g dry weight expressed as pyrocatechol equivalents. Then the suitability of CFE as an antioxidant was determined in peanut oil, and the decrease of lipid oxidation was monitored using thiobarbituric acid-reactive substances (TBARS) assay. CFE treatment significantly (P < 0.05) reduced lipid oxidation in peanut oil compared to the control. No significant differences (P = 0.05) in lipid oxidation were detected between CFE antioxidant and BHT antioxidant samples.     

Neuroprotective potential of some terebinth coffee brands and the unprocessed fruits of Pistacia terebinthus L. and their fatty and essential oil analyses

In the current study, neuroprotective effects of the ethyl acetate and methanol extracts of four terebinth coffee brands and the fruits of Pistacia terebinthus L. were investigated through enzyme inhibition tests against acetylcholinesterase, butyrylcholinesterase, and tyrosinase as well as antioxidant test systems. Antioxidant activity was measured using radical scavenging activity tests and metal-related tests including metal-chelation capacity, ferric-reducing antioxidant power (FRAP), and phosphomolybdenum reducing power (PRAP). The fatty oils of the coffee brands and the fruits and the fruit essential oil were examined by GC–MS. Total phenol and flavonoid contents were calculated spectrophotometrically. The extracts had moderate inhibition against butyrylcholinesterase (9.78–45.74% at 200 μg mL⁻¹) and potent scavenging activity against DPPH. They exerted strong activity in FRAP and metal-chelation tests and modest activity in PRAP test. Oleic acid was identified as the major fatty acid in the fatty oils, while α-pinene (26.31%) was dominant in the essential oil.     

The effect of diets supplemented with thyme essential oils and rosemary extract on the deterioration of farmed gilthead seabream (Sparus aurata) during storage on ice

The effect on quality were assessed for gilthead seabream fed five different diets: control (basal diet); BHT (basal diet with 200 mg kg⁻¹ of butylated hydroxytoluene); rosemary (basal diet with 600 mg kg⁻¹ of rosemary extract - Rosmarinus officinalis); carvacrol (basal diet with 500 mg kg⁻¹ of essential oil of Thymbra capitata, carvacrol chemotype); and thymol (basal diet with 500 mg g⁻¹ of essential oil of Thymus zygis, subspecies gracilis, thymol chemotype). After 18 weeks of experimentation, the animals were stored on ice at 4 °C for 0, 7, 14, and 21 days. Physical-chemical, microbiological and sensory analyses were carried out at each sampling point to determine the degree of deterioration in the gilthead seabream. Lower indices of oxidation were observed in animals who were administered feeds supplemented with BHT, carvacrol and (to a lesser degree) rosemary. Lower bacteria counts were observed for the BHT and thymol groups, in addition to a slower deterioration in terms of sensory perception. Accordingly, the addition of natural antioxidants to the diet may have an added effect on fish quality, delaying post mortem deterioration.     

Integrated management of insect vectors of Aspergillus flavus in stored maize, using synthetic antioxidants and natural phytochemicals

The purpose of this study was to investigate the insecticidal activity of two benzoic acids 2(3)-tert-butyl-4 hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT); two phenolic acids 3-phenyl-2-propenoic acid (CA) and trans-4-hydroxy-3-methoxycinnamic acid (FA) and two essential oils of Eugenia caryophyllata (clove tree) and Thymus vulgaris (thyme) against Sitophilus zeamais, Tribolium confusum and Rhyzopertha dominica, vector carriers of aflatoxigenic fungi in stored maize. The susceptibility of insects, the frequency of isolation of Aspergillus section Flavi in insects and maize, and the analysis of aflatoxin B₁ in maize were determined. BHA, BHT, BHA/BHT mixture and the natural phytochemicals AF and AF/AC mixture showed the highest insecticidal activity against S. zeamais, T. confusum and R. dominica after 120 days of incubation. The insecticidal efficacy of the volatile fraction of essential oils of clove and thyme showed less inhibition. There was no contamination of Aspergillus section Flavi in dead and live insects collected from maize treated with BHA. No aflatoxin B₁ accumulation was detected in the control and treatments. The information obtained shows that these substances have the potential to control pest insect vectors of aflatoxigenic fungi in stored maize in microcosms during 120 days.     

Major chemotypes and antioxidative activity of the leaf essential oils of Cinnamomum osmophloeum Kaneh. from a clonal orchard

Essential oils of 92 cutting clones from a clonal orchard of Cinnamomum osmophloeum Kaneh. were obtained by hydrodistillation and characterised by gas chromatography–mass spectrometry. Our results showed that the yields of essential oils ranged between 0.09% and 2.65% (vol/fresh wt). The constituents of essential oils varied among samples. The major chemotypes classified in the individual cutting clones were cinnamaldehyde (50 plants, representing 50–95% of the total volatiles), linalool (1 plant, 73.3%), β-cubebene (2 plants, 59.4% and 78.7%), and cinnamyl acetate (1 plant, 61.8%). The antioxidant activities of the four chemotypes were determined using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The antioxidant activities of the essential oil decreased in the order of cinnamyl acetate > cinnamaldehyde > β-cubebene > linalool. Indigenous cinnamon oil extract showed a good free radical-scavenging capacity at all concentrations studied, except at 2 μg/ml. The scavenging activity increased with increasing concentration of the extract. The capability of the four essential oil chemotypes to reduce the stable radical, DPPH, to DPPH-H was assayed by a decrease in the IC₅₀ values of 10.4 (cinnamyl acetate type) to 29.7 (linalool type) μg/ml. These results suggest that the leaf essential oil of C. osmophloeum possesses chemical compounds with antioxidant activity which can be used as natural preservatives in food and/or by the pharmaceutical industry. Trees in this plantation which can be used for further propagation for the production of chemotypes of interest were identified.     

Extraction optimisation,purification and major antioxidant component of red pigments extracted from Camellia japonica

The optimised extraction conditions of red pigments (RP) from Camellia japonica obtained with an orthogonal design L₉(3⁴) were solid/liquid ratio, temperature, pH and extraction time as 1/10, 60 °C, 1.5 and 4 h, respectively. The RP were then purified by the macroporous resin method, which showed the resin LX-68 was appropriate for purifying the pigments from C. japonica. The antioxidant activities of these pigments were also investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical in vitro model systems. The DPPH scavenging activity of pigment extract was comparable to that of standard butylated hydroxyanisole (BHA). The IC₅₀ values of the RP and BHA were 4.55 and 4.17 μg ml⁻¹, respectively. The pigments showed higher hydroxyl radical-scavenging activities than that of mannitol at the same concentration. Following activity-oriented separation, (−)-epicatechin was isolated as an active principle, which exhibited excellent DPPH free radical scavenging activities with IC₅₀ 5.08 μg ml⁻¹.     

Antioxidant and antibacterial activities of Nephelium lappaceum L. extracts

Ether, methanolic and aqueous extracts of lyophilized rambutan (Nephelium lappaceum L.) peels and seeds were evaluated for phenolic contents, antioxidant and antibacterial activities. High amounts of phenolic compounds were found in the peel extracts and the highest content was in the methanolic fraction (542.2 mg/g dry extract). Several potential antioxidant activities, including reducing power, β-carotene bleaching, linoleic peroxidation and free radical scavenging activity, were evaluated. The peel extracts exhibited higher antioxidant activity than the seed extracts in all methods determined (P < 0.05). The methanolic fraction was found to be the most active antioxidant as shown by their 50% DPPH inhibition concentration, 4.94 μg/mL. The results indicated this fraction exhibited greater DPPH radical scavenging activity than BHT and ascorbic acid (0.32 g dry extract/g BHT or ascorbic acid). Antibacterial activity against eight bacterial strains was assessed by disc diffusion and broth macrodilution methods. All peel extracts exhibited antibacterial activity against five pathogenic bacteria. The most sensitive strain, Staphylococcus epidermidis, was inhibited by the methanolic extract (MIC 2.0 mg/mL).     

Chemical composition and biological activities of Algerian Thymus oils

The compositions of essential oils isolated from nine samples of three Thymus species (Thymus algeriensis, Thymus pallescens and Thymus dréatensis) were analysed by GC and GC–MS, and a total of 114 components were identified. T. pallescens collected from various regions showed a great similarity in their compositions and were characterised by carvacrol (44.4–57.7%), p-cymene (10.3–17.3%) and γ-terpinene (10.8–14.2%) as the major components for four samples; only one sample was thymol-rich (49.3%) with a small amount of carvacrol (9.0%). On the other hand, T. algeriensis showed a chemical polymorphism, even for samples from the same location, and two new chemotypes for this species were proposed. Oxygen-containing monoterpenes were the predominant class (76.3%) in T. dreatensis oil, with linalool (30.4%), thymol (20.2%) and geraniol (19.6%) as the principal constituents. The oils were screened for their possible antioxidant activities by four complementary assays, namely DPPH free radical scavenging, hydroxyl radical scavenging, inhibition of lipid peroxidation and reducing power. The two new chemotypes of T. algeriensis exhibited strong hydroxyl radical scavenging (IC₅₀ = 2.2–3.3 μg/ml), but were not or only slightly active against the other radicals and exhibited a weak reducing power. Despite their chemical similarity, T. pallescens oils sometimes produced significant differences in their antioxidant activities. The essential oils were also screened for their antimicrobial activity against five bacteria (three Gram-positive and two Gram-negative) and one yeast (Candida albicans). The tested essential oils showed antimicrobial activity against the microorganisms used, in particular against two important pathogens, C. albicans and Helicobacter pylori.     

Antioxidant and antiacetylcholinesterase activities of essential oils from Cymbopogon schoenanthus L. Spreng. Determination of chemical composition by GC–mass spectrometry and C NMR

Cymbopogon schoenanthus L. Spreng, is an aromatic herb consumed in salads and used to prepare traditional meat recipes in Tunisia. The chemical composition, antioxidant activities and acetylcholinesterase inhibitory properties of the essential oils from fresh leaves, dried leaves and roots collected from three different locations in southern Tunisia, were evaluated. Essential oils were analysed by GC–mass spectrometry and ¹³C NMR. The major components were limonene (10.5–27.3%), β-phellandrene (8.2–16.3%), δ-terpinene (4.3–21.2%) and α-terpineol (6.8–11.0%). Antioxidant activity was measured by DPPH assay. The results ranged from 36.0% to 73.8% (2 μl of essential oil per mL of test solution).     

Headspace-solid phase microextraction (HS-SPME) analysis of oxidized volatiles from free fatty acids (FFA) and application for measuring hydrogen donating antioxidant activity

Volatile compounds from thermally oxidized free fatty acids (FFA) at 93 °C for 200 min were analyzed by headspace-solid phase microextraction (HS-SPME)-gas chromatography (GC). Commercially available mixtures of FFA were used instead of selecting specific vegetable oils with various fatty acid compositions. As oxidation time increased, total volatiles and some individual volatiles including hexanal, 2-hexenal, 2-heptenal, 2,4-heptadienal, 2-octenal, and 2,4-decadienal increased linearly with 0.99 coefficient of determination (R²) at specific oxidation time against conjugated dienoic acid (CDA) or p-anisidine value (p-AV). Total volatiles showed the highest linearity (R²) of 0.99 and 2-heptenal showed the highest increasing slope in peak areas for 200 min oxidation. Not all oxidized volatiles increased linearly during oxidation. Availability of HS-SPME method for determining hydrogen donating antioxidant activity was tested using FFA containing serially diluted butylated hydroxytoluene (BHT) at 93 °C for 60 min. 2-Heptenal and total volatiles showed higher linearity against BHT concentration than hexanal. HS-SPME could be a useful method to determine the hydrogen donating antioxidant activity from FFA using total volatiles or 2-heptenal as oxidation markers.     

Antioxidant potential of solvent extracts of Kappaphycus alvarezii (Doty) Doty – An edible seaweed

Various solvent extracts of Kappaphycus alvarezii, an edible red seaweed (family Solieriaceae) were screened for total phenol content and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferrous ion chelating activity, reducing power and antioxidant activity assays in a linoleic acid system with ferrothiocyanate reagent (FTC). The total phenol content of different extracts of K. alvarezii varied from 0.683 ± 0.040% to 2.05 ± 0.038%. The radical-scavenging activity of ethanol extract was, as IC₅₀ 3.03 mg ml⁻¹, whereas that of the water extract was IC₅₀ 4.76 mg ml⁻¹. Good chelating activity was recorded for methanol extract (IC₅₀ 3.08 mg ml⁻¹) wherein 67.0 ± 0.924% chelation was obtained using 5.0 mg ml⁻¹ of extract. The reducing power of the samples was in the following order: BHT > methanol > ethanol > ethyl acetate > water > hexane. But, in the linoleic acid system, the ethanol extract proved superior to the synthetic antioxidants butylated hydroxytoluene (BHT). Hence, these extracts could be considered as natural antioxidants and may be useful for curing diseases arising from oxidative deterioration.     

Quantitative determination of the phenolic antioxidants using voltammetric techniques

Synthetic antioxidants, such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butylhydroquinone (TBHQ), show sensitivity to voltammetric waves. The waves of these antioxidants, however, are seriously overlapped and it is difficult to determine them simultaneously. The influence of different parameters (working electrode, supporting electrolyte, pH, voltammetric technique) was evaluated in a quantitative simultaneous determination of three antioxidants in alcoholic mixtures and real sample foods. Glassy carbon (GC) and platinum (Pt) working electrodes were investigated as mediators of oxireduction reactions. Two supporting electrolytes were investigated: Britton-Robinson 0.1 mol l⁻¹ buffer (pH 2.0) and HCl 0.1 mol l⁻¹ (pH 2.0) both with 2 g l⁻¹ (p/v) of methanol. In this paper, voltammetric conditions for the analysis of up to three-component mixtures of antioxidant present at levels: 2.0-100.0 mg l⁻¹ for BHA, 4.0-100.0 mg l⁻¹ for TBHQ and 2.0-20.0 mg l⁻¹ for BHT at GC in HCl 0.1 mol l⁻¹ and 8.0-120.0, 10.0-130.0 and 4.0-30.0 mg l⁻¹ for BHA, TBHQ and BHT, respectively, at Pt in the same supporting electrolyte. The results show that for real food samples, the parameters investigated were satisfactory for quantitative determination using square wave voltammetry (SWV) without chemometric approaches and without suffering overlapping problems.     

Silphium L. extracts – composition and protective effect on fatty acids content in sunflower oil subjected to heating and storage

The influence of ethanol and hexane extracts from leaves, inflorescences, and rhizomes of Silphium perfoliatum, Silphium trifoliatum, Silphium integrifolium on fatty acid content changes in sunflower oil subjected to heating and storage was studied in comparison to the synthetic antioxidant – butylated hydroxyanisole (BHA). A positive effect of extracts made of above-ground and underground organs of Silphium on the durable quantitative composition of fatty acids was proven. Tested extracts elevated the value of change inhibition with reference to linoleic acid to a level comparable with BHA, and sometimes, in appropriate systems, they were characterized by better values (for oil stored for 180 days at room temperature, the inhibition coefficient for linoleic acid changes reached 4.6% for 0.04% BHA, and 7.09% for hexane extract made of S. trifoliatum inflorescences, 400 μl/2 g; for oil heated for 120 h, the inhibition coefficient of linoleic acid changes amounted to 11.32% for 0.06% BHA, and 15.69% for hexane extract made of S. perfoliatum rhizomes, 600 μl/2 g). It was found that active substances groups such as phenolic acids, flavonoids and terpenes were present in tested extracts.