A correlation between the level of phenolic compounds and the antioxidant capacity in cooked-with-rice and vegetable soybean (Glycine max L.) varieties

This study was undertaken to determine the content of phenolic compounds contained in embryo, cotyledon and seed coat of nine soybean (Glycine max L.) varieties. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity was also measured to determine the correlation between the level of phenolic compounds and antioxidant capacity. A total of 10 anthocyanin constituents and 21 phenolic compounds was detected and quantified. In all nine varieties, the seed coat and cotyledon had the highest and lowest levels of phenolic compounds and anthocyanins, respectively despite the fact that all of them showed a wide variation in total amounts of phenolic compounds and anthocyanins in seed coat, embryo, and cotyledon. The seed coat tissue, but not other seed parts, showed a strong correlation between the seed coat color and the content of both phenolic compounds and anthocyanins. The brown and black soybean seed coat contained much higher levels of phenolic compounds and anthocyanins in seed coat tissue than the yellow or green coat soybean. Among the individual phenolic compounds, syringic acid (214 μg g⁻¹) and chlorogenic acid (31 μg g⁻¹) were highest in seed coat and embryo, respectively. Myricetin was highest both in whole seed (16.7 μg g⁻¹) and cotyledon (16.0 μg g⁻¹), being equivalent to 20 and 30% of total phenolic compounds, respectively. Among the 10 anthocyanins, cyanidin-3-glucoside was found to accumulate at the highest level in the seed coat (1783 μg g⁻¹), whole seed (106 μg g⁻¹) and embryo (0.35 μg g⁻¹), which correspond to 95, 96, and 40% of the total anthocyanin contents, respectively. The cotyledon accumulated pelargonidin-3-glucoside (0.39 μg g⁻¹) at the highest level that is equivalent to 62% of total anthocyanin contents. DPPH activity was found to have a strong correlation (^***probability < 0.001) with phenolic compounds (0.67^***) and anthocyanins (0.70^***).J. A. Kim, W. S. Jung, and S. C. Chun are equally contributed to this work.     

Effects of baking on cyanidin-3-glucoside content and antioxidant properties of black and yellow soybean crackers

Black soybean is a potential functional food ingredient with high anthocyanin content, but the ability to maintain anthocyanin content under dry heat processing has not been reported. This study investigated the effects of soybean seed coat colour and baking time–temperature combinations on the extractable antioxidant properties of a soy cracker food model. Crackers prepared with black soybeans had significantly higher TPC, total isoflavones, and peroxyl, hydroxyl, and ABTS⁺ radical scavenging abilities than their yellow counterparts, at all time–temperature combinations. Cyanidin-3-glucoside (C3G) was detected only in black soybean crackers, and all baking treatments significantly decreased C3G. The greatest losses occurred at the low temperature × long time and high temperature × short time, the smallest loss with moderate temperature × short/medium time. The high temperature treatment altered phenolic acid and isoflavone profiles; however, total isoflavones were unaffected. Overall results suggest that moderate baking temperature at minimal time may best preserve anthocyanin and other phenolics in baked black soybean crackers.     

Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component

Buddleia officinalis Maxim, commonly used as rice dye for festivals, was extracted with ethanol using microwave-assisted extraction and Soxhlet extraction. The antioxidant activities of microwave-assisted extract of B. officialis (MEB) and Soxhlet extract of B. officianils (SEB) at the optimum extraction conditions were evaluated and compared with synthetic antioxidant butylated hydroxytoluene (BHT) employing DPPH free radical assay, ABTS assay, total antioxidant activity and reducing power. MEB and SEB had stronger antioxidant activities than BHT in all assays except reducing power, and the effects decreased as follows: MEB > SEB > BHT. The total phenolic contents of MEB and SEB reached 113.56 mg/g and 100.94 mg/g dry weight of extract, respectively, expressed as pyrocatechol equivalents, while the total flavonoids contents were 75.33 mg/g and 62.56 mg/g dry weight of extract, respectively, expressed as catechin equivalents (P < 0.05). Higher phenolic and flavonoids compounds may be major contributors to their higher antioxidant activities. Following activity-oriented separation, luteolin was isolated as an active principle, which exhibited excellent free radical scavenging activities with DPPH IC₅₀ 3.09 μg/ml and ABTS IC₅₀ 2.20 μg/ml.     

Antioxidant effect of oregano (Lippia berlandieri v. Shauer) essential oil and mother liquors

The conventional steam distillation process for oregano (Lippia berlandieri v. Shauer) essential oil extraction produces large volumes of mother liquor. This residual liquid represents a potential value because the soluble antioxidants it contains. Essential oil and ethyl acetate mother liquor extracts (MLEs) were evaluated for antioxidant activity. Total phenolic content and antioxidant activities by the 2-2′-diphenyl-1-picrylhydrazyl (DPPH) method, by the deoxyribose degradation assay, and by oxidation of low density lipoproteins (LDL) with CuSO₄ were evaluated. Oil yield was 4.34%. Total phenolic content was 151 ± 2.00 and 150.5 ± 0.98 mg of GAE (gallic acid equivalents)/mL for the essential oil and MLEs, respectively. DPPH assay showed a low radical scavenging activity (RSA) for oregano essential oil. Meanwhile MLEs exhibited no significant RSA at low concentrations, but at higher concentrations (100 μg/mL), it was superior to those exhibited by the controls ascorbic acid and butylated hydroxytoluene (BHT). Deoxy-d-ribose assay results for both essential oil and MLEs showed a good hydroxyl radical RSA at the concentrations tested. Essential oil and MLEs delayed induction time effectively. Solubility problems, chemical constituents, and their hydrophilic–lipophilic distribution are key factors that explain samples behavior for an eventual use of these natural products.     

Standardised Mangifera indica extract is an ideal antioxidant

The standardised ethanolic and aqueous extracts of Mangifera indica leaf were prepared and analysed for their free radicals scavenging activity. The IC₅₀ values using the DPPH assay were 0.17 ± 0.02 and 0.49 ± 0.4 mg/ml, respectively. Standardised ethanolic extracts of the M. indica leaf had a solid content of 9.1 ± 0.7%, mangiferin concentration of 73 ± 0.17 mg/g of dry weight of the extract, free radical scavenging activity (IC₅₀) of 0.17 ± 0.02 mg/ml and total phenolic content of 590 ± 48 mg/g of extract. The protection exhibited by these extracts against lipid peroxidation was superior to butylated hydroxytoluene (BHT) and commercial grape seed extract. These extracts at higher concentration did not exhibit pro-oxidant activities when compared to vitamin C. Our findings also show that the aqueous and ethanolic extracts of M. indica leaf protect NIH/3T3 cells from oxidant-induced cell death.     

Antioxidant activity and phenolics of an endophytic Xylaria sp. from Ginkgo biloba

The objective of this study was to evaluate the antioxidant activity of cultivated fruiting bodies of an endophytic Xylaria sp. (strain number YX-28), from Ginkgo biloba. The results indicated that the methanol extract exhibited strong antioxidant capacity in both 2,2-diphenyl-1-picrylhydrazyl (DPPH) analysis and β-carotene–linoleic acid model system. Total phenolic and flavonoid contents extracted by different solvents were determined using Folin–Ciocalteu procedure and the flavonoid–aluminium method. The results showed that total phenolic and flavonoid contents were the highest in methanol extract (54.51 ± 1.05 mg gallic acid equivalent/g dry weight and 86.76 ± 0.58 mg rutin equivalent/g dry weight), while the hexane extract was the lowest (9.71 ± 0.57 mg GAE/g dw and 10.14 ± 0.76 mg RE/g dw, respectively). The correlation coefficients from regression analysis showed a positive relationship between total phenolic content in the extracts and DPPH activity (R² = 0.7336), as well as between total flavonoid content and DPPH activity (R² = 0.9392). Furthermore, GC/MS method was used to confirm the presence of phenolics with antioxidant activity in the methanol extract and resulted in the identification of 41 compounds, esters, phenolics, alkanes, carboxylates and alcohols being the main components. In conclusion, cultivated fruiting bodies of Xylaria sp.YX-28 may have potential as natural antioxidant.     

Antioxidative and anti-inflammatory properties of Citrus sulcata extracts

Citrus sulcata was subjected to ultrasound, high-pressure, and Soxhlet extractions. The antioxidant and anti-inflammatory activities of the extracts were evaluated qualitatively and quantitatively. The antioxidant content of the peel extract was twice that of the fruit extract. The quantitative analysis showed that the narirutin and hesperidin contents in the peel extracts were 8.8 and 7.5 mg/100 g, respectively. These extracts had a total phenolic content of 112.22 ± 2.89 gallic acid equivalent (GAE) mg/100 g, a total flavonoid content of 54.09 ± 1.01 rutin equivalent (RE) mg/100 g, DPPH radical scavenging activity of 46%, and antioxidant activity of 213.25 ± 2.82 μM of Trolox equivalents (TEAC). C. sulcata extracts could prevent hydrogen peroxide (H₂O₂)-induced oxidative stress, reduce expression of the inflammatory markers nuclear Factor kappa B (NFκB) and phosphorylated IκBα (p-IκBα) in A549 human lung carcinoma cells, and inhibit Lipopolysaccharide (LPS)-induced THP-1 monocyte differentiation to an extent of 85%.     

Chemical composition,anthocyanins, non-anthocyanin phenolics and antioxidant activity of wild bilberry (Vaccinium meridionale Swartz) from Colombia

Berries of Vaccinium meridionale Swartz native to Colombia were analysed for chemical composition, total phenolic content, anthocyanin content, and antioxidant activity. In addition, high-performance liquid chromatography with photodiode array detection (HPLC–DAD) and HPLC-electrospray ionisation tandem mass spectrometry (ESI–MS/MS) were used to determine anthocyanin and phenolic composition. Anthocyanin content was 329.0 ± 28.0 mg cyanidin 3-glucoside equivalents/100 g (fresh weight) FW and total phenolic content was 758.6 ± 62.3 mg gallic acid equivalent/100 g FW. Cyanidin 3-galactoside was the major anthocyanin while the most abundant non-anthocyanin phenolic was chlorogenic acid.     

Antioxidant capacity of the extracts from pulp of Osmanthus fragrans and its components

In this study, a significant inhibitory effect (EC₅₀ = 50.29 μg/ml) on the ABTS free radicals was detected in the crude ethanol extracts (CEE) of Osmanthus fragrans' pulp. Then, CEE was separated into methanol soluble fraction (MSF) and a methanol insoluble fraction (MISF), and their scavenging activities on the ABTS free radicals ascended in the order: MSF > CEE > MISF, respectively. Accordingly, the total phenolic (TP) contents of MSF, CEE and MISF were 40.53 ± 0.27, 35.37 ± 0.18 and 29.62 ± 0.23 mg/g (dry weight), respectively. Furthermore, salidroside (tyrosol 8-O-β-glucopyranoside), which showed powerful antioxidant activities in all test system was isolated from MSF. The antioxidant activity of salidroside (81.54 μg/ml) on DPPH free radicals was found to be nearly twice of that of MSF (144.78 μg/ml), and its total antioxidant activity is superior to that of the synthetic antioxidant butylated hydroxytoluene (BHT). In conclusion, our results suggested that O. fragrans' pulp, which was often considered as agricultural wastes, could be a promising source of natural antioxidants, and that MSF and salidroside may be good candidates for further development as natural antioxidants.     

Soluble phenolics and antioxidant properties of soybean (Glycine max L.) cultivars with varying seed coat colours

This research was the first to investigate nutritional components, including soluble phenolics (isoflavones and anthocyanins), protein, oil, and fatty acid as well as antioxidant activities in different coloured seed coat soybeans (yellow, black, brown, and green) for two crop years. The soluble phenolics differed significantly with cultivars, crop years, and seed coat colours, while protein, oil, and fatty acid exhibited only slight variations. Especially, malonylgenistin and cyanidin-3-O-glucoside compositions had the most remarkable variations. Green soybeans had the highest average isoflavone content (3079.42 μg/g), followed by yellow (2393.41 μg/g), and black soybeans (2373.97 μg/g), with brown soybeans showing the lowest value (1821.82 μg/g). Anthocyanins showed only in black soybeans, with the average contents of the primary anthocyanins, cyanidin-3-O-glucoside, delphinidine-3-O-glucoside, and petunidin-3-O-glucoside, quantified at 11.046, 1.971, and 0.557 mg/g, respectively. Additionally, Nogchae of green soybean and Geomjeongkong 2 of black soybean may be recommended as potential cultivars owing to the highest average isoflavone (4411.10 μg/g) and anthocyanin (21.537 mg/g) contents. The scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals also differed remarkably, depending upon isoflavone and anthocyanin contents, with black soybeans exhibiting the highest antioxidant effects.     

Antioxidant activity of white and black sesame seeds and their hull fractions

The total phenolic content (TPC), total antioxidant status (TAS), free radical scavenging capacity, inhibition of low density lipoprotein (LDL) cholesterol and metal chelating capacity of extracts of whole black and whole white sesame seeds and their hull fractions in 80% aqueous ethanol were investigated. The TPC of whole black sesame seeds and hull extracts were 29.9 ± 0.6 and 146.6 ± 0.6 mg catechin equivalents/g crude ethanolic extract, respectively. The corresponding values for white sesame were 10.6 ± 1.6 and 29.7 ± 0.9 mg catechin equivalents/g crude ethanolic extract. The TAS as determined by Trolox equivalent antioxidant capacity assay and expressed as Trolox equivalents was highest for black sesame hulls (65.9 ± 1.7) while white seeds showed the lowest (4.4 ± 0.6). Free radical scavenging capacity of sesame extracts (5–40 μg/mL) was measured using 2,2-diphenyl-1-picrylhydrazyl radical. The highest scavenging capacity was obtained at 40 μg/mL and was 94.9 ± 0.8, 25.1 ± 0.4, 14.4 ± 0.9 and 2.5 ± 0.4 for black sesame hulls, black sesame seeds, white sesame hulls and white sesame seeds, respectively. Inhibition of LDL oxidation at 100 ppm level was highest for black sesame hulls (96.7%) followed by those for white sesame hulls (84.6%), black sesame (78.4%) and white sesame seeds (57.3%). Sesame products displayed good ferrous ion chelating capacities, which ranged from 12% to 46% and 17% to 62% at 50 and 100 ppm levels, respectively. Results demonstrated considerable antioxidant activity of sesame products tested especially black sesame hulls.     

Assessment of in vitro antioxidant capacity and polyphenolic composition of selected medicinal herbs from Leguminosae family in Peninsular Malaysia

Determination of total phenolic (TPC), flavonoid and anthocyanin contents, and various antioxidant activities (2,2-diphenyl-1-picrylhydrazyl radical scavenging, ferric reducing power, ferrous ion chelating and lipid peroxidation inhibition) of leaves and flowers of Bauhinia kockiana, Caesalpinia pulcherrima and Cassia surattensis were performed in this study. The B. kockiana flower was found to possess the highest TPC (8280 ± 498 mg GAE/100 g), free radical scavenging activity (ascorbic acid equivalents 14,600 ± 2360 mg AA/100 g) and reducing ability (72.4 ± 8.7 mg GAE/g). Rutin and chlorogenic acid were detected in the plants, where the C. pulcherrima leaf contained the highest amount of rutin (669 ± 26 mg/100 g), while minute amounts of chlorogenic acid were detected in C. surattensis leaf (9.13 ± 0.44 mg/100 g). The C. pulcherrima leaf displayed the highest ferrous ion chelating and lipid peroxidation inhibition activities. Positive correlation was observed between TPC and various antioxidant activities.     

Anti-oxidative analysis,and identification and quantification of anthocyanin pigments in different coloured rice

Anthocyanin pigments in coloured rice cultivars were isolated and identified using high-performance liquid chromatography techniques. Two black rice cultivars (Asamurasaki, Okunomurasaki) contained three major anthocyanins: cyanidin-3-glucoside, peonidin-3-glucoside and malvidin. Chinakuromai (black) rice additionally contained a fourth anthocyanin, petunidin-3-glucoside. Four red rice cultivars contained only malvidin. The total anthocyanin content varied greatly among black rice cultivars (79.5–473.7 mg/100 g), but was lower in red rice (7.9–34.4 mg/100 g). Total phenolic content was similar between red (460.32–725.69 mg/100 g) and black (417.11–687.24 mg/100 g) rice. The oxygen radical absorbing capacity was ranked as follows: red (69.91–130.32 μmol Trolox/g) > black (55.49–64.85 μmol Trolox/g) > green (35.32 μmol Trolox/g) > white (21.81 μmol Trolox/g) rice. The antioxidant capacity resulted mainly from the seed capsule, not the endosperm. The anthocyanin pigments contributed little to the total antioxidant capacity of red (0.03–0.1%) and black (0.5–2.5%) rice cultivars. Hence, the antioxidant capacity is derived mainly from other phenolic compounds.     

Rind of the rambutan, Nephelium lappaceum, a potential source of natural antioxidants

The rind of rambutan, which is normally discarded was found to contain extremely high antioxidant activity when assessed using several methods. Although having a yield of only 18%, the ethanolic rambutan rind extract had a total phenolic content of 762 ± 10 mg GAE/g extract, which is comparable to that of a commercial preparation of grape seed extract. Comparing the extract’s pro-oxidant capabilities with vitamin C, α-tocopherol, grape seed and green tea, the rind had the lowest pro-oxidant capacity. In addition, the extract at 100 μg/ml was seen to limit oxidant-induced cell death (DPPH at 50 μM) by apoptosis to an extent similar to that of grape seed. The extracts were not cytotoxic to normal mouse fibroblast cells or splenocytes while the powderised rind was seen to have heavy metals contents far below the permissible levels for nutraceuticals. Our study for the first time reveals the high phenolic content, low pro-oxidant capacity and strong antioxidant activity of the extract from rind of Nephelium lappaceum. This extract, either alone or in combination with other active principles, can be used in cosmetic, nutraceutical and pharmaceutical applications.     

Phenolic content and antioxidant activity of cantaloupe (cucumis melo) methanolic extracts

The objectives of this study were to determine phenolic content and antioxidant activity of methanolic extracts from different parts of cantaloupe (leaf, stem, skin, seed and flesh). The flesh extract afforded the highest yield (89.6 ± 0.3%) whilst the lowest yield was obtained from the seed (13.7 ± 0.5%) (p < 0.05). The leaf extract showed the highest total phenolic content (26.4 ± 0.3 mg GAE/g extract) and total flavonoid content (69.7 ± 3.37 μg RE/g extract) accompanied with best antioxidant activity through all antioxidant assays (p < 0.05). In addition, the stem extract also exhibited good antioxidant activity. Thus, these results suggest that methanolic extracts of cantaloupe leaf and stem may serve as a potential source of natural antioxidant for food and nutraceutical application.     

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.     

Variation of active constituents and antioxidant activity in pyrola [P. incarnata Fisch.] from different sites in Northeast China

The variation of antioxidant activity and active components in pyrola [Passiflora incarnata Fisch.] from eight sites in Northeast China were investigated. Total phenolic and flavonoid contents were determined and varied within the range of 39.66–181.48 mg/g and 2.47–22.11 mg/g, respectively. Antioxidant activities were determined by scavenging activity against DPPH and ABTS, by a reducing power test and by a β-carotene-linoleic acid bleaching test. The IC₅₀ of Tahe samples determined by the DPPH test was 0.106 ± 0.006 mg/mL which was very close to that of Vc (0.076 ± 0.004 mg/mL). The Tahe samples had good antioxidant activity. Principal component activity analysis indicated that the Tahe samples of P. incarnata had the highest potential antioxidant properties, and may be a valuable antioxidant natural resource in the northeast of China.     

Antioxidant capacity and vasodilatory properties of Mediterranean food: The case of Cannonau wine,myrtle berries liqueur and strawberry-tree honey

The aim of this work was to use different assays to evaluate the antioxidant and vasodilatory properties of three typical food products from the Mediterranean area and to correlate these activities with their phenolic content. For this purpose, red wines Cannonau, liqueurs obtained by cold maceration of myrtle (Myrtus communis L.) berries and bitter honeys obtained from strawberry-tree flowers (Arbutus unedo L.) were analysed. The total phenolic (TP) content was measured spectrophotometrically with a modified Folin–Ciocalteau method and phenolic compounds were identified and dosed by HPLC-DAD and LC-MS/MS. Antioxidant activities were evaluated with DPPH, FRAP and ABTS assays and the in vitro vasodilatory effects were assessed using norepinephrine precontracted rat aortic rings. Cannonau wines and myrtle liqueurs showed high levels of TP (1978 ± 279 and 1741 ± 150 mg GAE/L, respectively), linearly correlated to the results of FRAP, ABTS, and DPPH assays. Their maximal vasodilatory activity was 61.7 ± 4.1% and 53.0 ± 3.0%, respectively. Although strawberry-tree honey contained relatively high levels of phenolic compounds (922 ± 38 mg GAE/kg), it did not induce vasodilation, even at the highest dose tested (0.206 g/L). These results indicate that foods with high levels of phenolic compounds should be studied using several different biological assays before being recommended to the general public as functional foods.     

Phenolic acid contents of kale (Brassica oleraceae L. var. acephala DC.) extracts and their antioxidant and antibacterial activities

Nine phenolic acids were identified and quantified by HPLC–MS in leaves and 10 in seeds of kale (black cabbage). The free, ester (methanol-soluble), glycoside and ester-bound (methanol-insoluble) phenolic acid contents of the leaves were 487, 532, 4989 and 6402 ng/g fresh weight, respectively. Ferulic and caffeic acids (total contents; 4269 and 4887 ng/g, respectively) were the most abundant. The seed contents of these fractions were 1993, 1477, 1231 and 4909 ng/g dry weight (DW), respectively, and sinapic acid was the most abundant (5037 ng/g DW). The fractions’ total phenolic contents, determined colorimetrically, were highly correlated with their DPPH scavenging capacity, and in antimicrobial activity assays, with nine test organisms representing a wide array of taxa, all of the fractions were effective against Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis and (most strongly) Moraxella catarrhalis. Antimicrobial and antioxidant activities of kale phenolics in free and conjugated forms are discussed.     

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.