JOURNAL OF MEDICINAL FOOD J Med Food 17 (10) 2014, 1151–1157 # Mary Ann Liebert, Inc., and Korean Society of Food Science and Nutrition DOI: 10.1089/jmf.2013.0057

Antioxidant Activity, Total Phenolic and Flavonoid Contents of Some Medicinal and Aromatic Plants Used as Herbal Teas and Condiments in Iran Abdollah Ghasemi Pirbalouti,1,2 Amir Siahpoosh,3 Milad Setayesh,3 and Lyle Craker 2 1

Research Center for Medicinal Plants & Ethno-veterinary, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran. 2 Medicinal Plants Program, Stockbridge School of Agriculture, College of Natural Science, University of Massachusetts, Amherst, Massachusetts, USA. 3 Department of Pharmacognosy, Herbal Medicine and Natural Products Research Center, School of Pharmacy, Ahwaz Jondishapur University of Medical Sciences, Ahwaz, Iran. ABSTRACT Total phenolic and flavonoid contents and antioxidant activity of four Iranian herbs (Lamiaceae) were investigated. The antioxidant activity of methanol extracts of thyme (Thymus daenensis Celak.), Bakhtiari savory (Satureja bachtiarica Bung.), dragonhead (Dracocephalum multicaule Montbr & Auch), and woundwort (Stachys lavandulifolia Vahl.) was evaluated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing/antioxidant power (FRAP), and trolox equivalent antioxidant capacity (TEAC). A comparison of all plant extracts in the DPPH assay indicated that dragonhead and thyme were the most effective free radical scavenging agents. Thyme demonstrated a relatively strong antioxidant activity in both the FRAP and TEAC assays. The total phenolic content of all the extracts ranged from 99 to 208 mg TAE/g extract with thyme exhibiting the highest phenolic content. The flavonoid content of the extracts, which ranged from 10.1 to 22.2 rutin equivalents/g of extract, was highest in dragonhead. A positive correlation was noted between the total phenolic content and antioxidant activity in both the FRAP and TEAC assays, while no significant correlation was observed between the DPPH, TEAC, and FRAP assay and total flavonoid, suggesting that the level of antioxidant activity in these plants varies greatly, but the total phenolic in the plant extracts provided a substantial antioxidant activity. Experimental results indicate that thyme and dragonhead extracts could be an important dietary source of phenolic compounds with high antioxidant capacity.

KEY WORDS:  DPPH  dragonhead  FRAP  savory  thyme  TEAC  woundwort

tannins), nitrogen compounds (alkaloids, amines, and betalains), vitamins, terpenoids, and other endogenous metabolites that have the antioxidant activity.6–12 For this reason, herbs and their constituents have been used in the food industry for their flavoring and biological activities since ancient times.13 Recently, the interest in finding naturally occurring antioxidants to replace synthetic antioxidants in foods and medicines has increased considerably, primarily due to the possible carcinogenicity of the synthetic antioxidants.9 In the present study, the antioxidant activity of extracts from four Iranian herbs used as herbal tea and condiments, including thyme (Thymus daenensis subsp. daenensis Celak.), savory (Satureja bachtiarica Bung.), dragonhead (Dracocephalum multicaule Montbr & Auch), and woundwort (Stachys lavandulifolia Vahl.) growing wild in Iran, was evaluated. T. daenensis, an endemic species of Iran, grows at high altitudes in the Zagros Mountains.14 T. daenensis is a traditional medicinal herb, the infusions and decoctions of aerial parts are used for the treatment of colds and digestive problems.15,16 S. bachtiarica is a perennial aromatic herb distributed in Zagros mountain range, southwestern Iran.

INTRODUCTION

N

umerous physiological and biochemical processes in the human body may produce oxygen-centered free radicals and other reactive oxygen species as by-products.1 Overproduction of such free radicals can cause oxidative damage to bimolecular constituents in the body, such as lipids, proteins, and DNA, which eventually leads to chronic diseases, such as atherosclerosis, cancer, diabetes, aging, and other degenerative diseases.2–4 Deterioration of lipids is catalyzed by internal and external factors, such as free radicals, metal ions, light, and heat.5 Oxidation, deterioration, and microbial contamination that occur in food products can lead to sickness in consumers and economic loss to processors.5 Herbs can contain a wide variety of free radical scavenging molecules, such as phenolic compounds (phenolic acids, flavonoid, quinones, coumarins, lignans, stilbenes, and Manuscript received 25 February 2013. Revision accepted 27 April 2014 Address correspondence to: Abdollah Ghasemi Pirbalouti, PhD, Research Center for Medicinal Plants & Ethno-veterinary, Shahrekord Branch, Islamic Azad University, PO. Box: 166, Shahrekord, Iran, E-mail: [email protected] or [email protected]

1151

1152

GHASEMI PIRBALOUTI ET AL.

The infusions and decoctions of aerial parts of S. bachtiarica are used for the treatment of colds and also as an analgesic and antiseptic by the Bakhtiari and Chaharmahali tribes of Iran.16 D. multicaule, which generally grows at high altitudes in Zagros Mountains of Southwest and Northwest Iran,14 is commonly used by the Bakhtiari and Chaharmahali tribes as a folk medicine for treatment of sleeplessness, pain, and infections.17 S. lavandulifolia, is a plant native of Iran from which the aerial parts are used in traditional medicinal as herbal tea for treatment of gastrointestinal disorders, inflammatory diseases, anxiety, coughs, ulcers, and other problems.15,18 To our knowledge, no documented reports on the antioxidant activity of the methanol extracts of T. daenensis, S. lavandulifolia, D. multicaule, and S. bachtiarica using different antioxidant assays are available. MATERIALS AND METHODS Plant material Thyme (T. daenensis subsp. daenensis Celak.), savory (S. bachtiarica Bung.), dragonhead (D. multicaule Montbr & Auch), and woundwort (S. lavandulifolia Vahl.) plants were collected from mountainous areas of Zagros, Chaharmahal va Bakhtiari district, Iran, between May and August, 2011 (Fig. 1). All collected specimens were dried and identified by comparison with authentic specimens deposited at the Herbarium of the Research Centre of Agriculture and Natural Resources of Chaharmahal va Bakhtiari (CHB) and I.A.U., Shahrekord (IAUSHK), Iran (Table 1).

Table 1. Iranian Herbs Used in This Study Voucher number

Scientific name Thymus daenensis Satureja bachtiarica Dracocephalum multicaule Stachys lavandulifolia

Parts used

Extract yield (%)

CHB.3092 CHB.1999

Areal parts Areal parts

4.70 – 1.23 8.80 – 3.44

IAUSHK.67

Areal parts

10.20 – 2.97

IAUSHK.73

Inflorescences

9.90 – 1.18

Chemicals and reagents FeCl3$6H2O, acetic acid, FeSO4$7H2O, NaC2H4O2$7H2O, FeCl2$7H2O tannic acid, methanol, and HCl (37%) used in this study were purchased from Merck Co. The 2,4,6-tri(2-pyridyl)-s-triazin (TPTZ) was purchased from Fluka Chemicals, rutin was obtained from Roth Co., and the Folin–Ciocalteu reagent, the 1,1-diphenyl-2-picrylhydrazyl (DPPH), and the ABTS radical cation [2,20 -azino-bis (3ethylbenzthiazoline-6-sulfonic acid)] were purchased from Sigma–Aldrich Co. Extract preparation The plant samples were subsequently air-dried for 5 days in a shaded room at 30C – 5C and then ground to a fine powder using a Moulinex food processor (Moulinex International). The ground powdered samples were subsequently passed through a 20-mesh sieve to remove large pieces of debris in preparation for essential oil extraction. The ground samples were subsequently dried to a constant weight over a desiccant (Na2SO4) at room temperature (30C). A 100-g sample was extracted with 1500 mL methanol at 25C for 48 h followed by shaking for 2 h. The methanol was subsequently removed under reduced pressure on a rotary evaporator at 40C. All prepared extracts were stored at 5C until use. Determination of total phenolic compounds The total amount of phenolic compounds in each extract was determined using the Folin–Ciocalteu method following the procedure of Singleton and Rossi19 with some modifications. Briefly, a 0.5 mL of the sample was mixed with 2.5 mL of Folin–Ciocalteu’s phenol reagent for 5 min at 37C, 2 mL of saturated Na2CO3 (7.5%) was added, and the mixture was brought to 10 mL with the addition of deionized distilled water. The mixture was maintained at room temperature in the dark for 120 min and then the absorbance was measured at 765 nm against a reagent blank using a Shimadzu UV–Vis (Shimadzu Corp.) spectrophotometer. Tannic acid was used as the reference standard and the total phenolic content was expressed as mg of tannic acid equivalents per gram of each extract on dry basis (mg TAE/g extract).

FIG. 1. The herbs tested, including Thymus daenensis Celak. (a), Stachys lavandulifolia Vahl. (b), Dracocephalum multicaule Montbr & Auch. (c), and Satureja bachtiarica Bung. (d). Color images available online at www.liebertpub.com/jmf

Estimation of flavonoid content The flavonoid content in extracts was determined spectrophotometrically according to the procedure of Lamaison

BIOACTIVITY OF SOME OF IRANIAN HERBS (LAMIACEAE)

1153

and Carnat,20 using a method based on the formation of a flavonoid–aluminum complex with a maximum absorbtivity at 430 nm. The flavonoid content was calculated using the following linear equation based on the calibration curve: A = 0.0224 C - 0.0133, r = 0.9993, where A is the absorbance and C is the flavonoid content in mg/g. Diluted samples (2 mL) were separately mixed with 2 mL of 2% AlCl3$H2O (w/v) and incubated at room temperature for 10 min before measuring the absorbance of the reaction mixture at 430 nm. The total flavonoid content is expressed as mg of rutin equivalents (RE) per g of extract. All replicates were performed in triplicate.

ethylbenzthiazoline-6-sulfonic acid)]. Total antioxidant activity values were estimated by the Trolox equivalent antioxidant capacity (TEAC) test.23 Briefly, ABTS was dissolved in water to a 7 mM concentration, and the ABTS radical cation (ABTS + ) was produced by reacting the ABTS stock solution with 2.45 mM potassium persulfate (final concentration) and allowing the mixture to stand in the dark at room temperature for 16 h before use. The ABTS + solution was diluted to get an absorbance of 0.700 – 0.020 at 734 nm. The percentage inhibition of absorbance at 734 nm was calculated using Equation 1. All determinations were done at least three times.

Determination of antioxidant activity

Statistical analysis

The ferric reducing/antioxidant power (FRAP) assay was used to determine the antioxidant activity, using the procedure described by Benzie and Strain.21 The FRAP reagent contained 2.5 mL of a 10 mM TPTZ solution in 40 mM HCl, 2.5 mL of 20 mM FeCl3$6H2O, and 25 mL of 300 mM acetate buffer (pH = 3.6). A standard curve was prepared using concentrations of 0.1–2 mM FeSO4$7H2O. The reaction mixture was incubated at 37C for 30 min, and the absorbance was measured at 593 nm using the above-described spectrophotometer. The antioxidant equivalent concentration (EC1 = 1) was defined as the concentration of antioxidant having a ferric-TPTZ reducing ability equivalent to that of 1 mM FeSO4$7H2O and represented the concentration of antioxidant equivalent to the theoretical absorbance value of a 1 mM concentration of Fe (II) solution. All replicates were performed in triplicate.

All assays were done in triplicates and results are expressed as the mean – standard deviation. The data were statistically analyzed in one-way analysis of variance using the program SPSS (17.0). Means of the scavenging activity, EC1, and IC50 of various extracts for different antioxidant assays were compared with the least significant different test at P £ .05 level. The Pearson correlation coefficient (r) and the probability value (P) were used to show correlation and their significance by using SPSS.

DPPH radical scavenging activity The DPPH radical scavenging activity of extracts was determined using the method proposed by Brand-Williams et al.22 Samples of selected concentrations of the plant extracts were mixed with an equal volume of 0.2 mM methanolic solution of DPPH. The disappearance of DPPH was followed spectrophotometrically at 515 nm beginning immediately after mixing and incubation for 1, 5, 15, and 30 min at room temperature. The absorbance of the DPPH radical without an antioxidant against a control measured daily. The control contained methanol instead of the antioxidant solution, while blanks contained methanol instead of DPPH solution. The amount of the sample necessary to decrease the absorbance of DPPH by 50% (IC50) was calculated graphically. The percentage inhibition was calculated according to Equation 1:   AC(0)  AA(t)  100, % inhibition ¼ AC(0)

RESULTS AND DISCUSSION Extraction yield Results of this study indicated differences in extract yields obtained from the studied species (Table 1). The highest extract yield (10.2%) was obtained from D. multicaule. An earlier study by Rabbani et al.24 reported that a hydroalcohol extract yield of woundwort was 11.3%. No information on extracts of the other tested plants is available. Phenolic content Phenolic compounds are believed to account for a major portion of the antioxidant capacity in many plants.25 A significant difference (P £ .05) for total phenolic content was measured among the four extracts examined. The total phenolic content of the various extracts varied from 99 to 208 mg TAE/g extract. The maximum total phenolic content was obtained for the extract of thyme (208 mg TAE/g extract) followed by dragonhead (180 mg TAE/g extract) (Table 2). The total phenolic content was similar in the extracts of savory and woundwort at 99.0 and 103 mg TAE/ g extract, respectively (Table 2). Total flavonoid content

Where AC(0) is the absorbance of the control at t = 0 min; and AA(t) is the absorbance of the antioxidant at t = 1, 5, 15, and 30 min. Trolox equivalent antioxidant capacity assay The free radical scavenging activity was determined using the stable ABTS radical cation [2,20 -azino-bis (3-

Flavonoids are widespread plant secondary metabolites, including flavones, flavanols, and condensed tannins. Epidemiological studies suggest that the consumption of flavonoidrich foods protects against human diseases associated with oxidative stress. In vitro, flavonoids from several plant sources have shown free radical scavenging activity and protection against oxidative stress.26,27 The flavonoid content

1154

GHASEMI PIRBALOUTI ET AL. Table 2. Antioxidant Activity, Total Phenolic Content, and Flavonoid Content of Methanol Extracts from Four Herbs (Lamiaceae)

Herbs T. daenensis S. bachtiarica D. multicaule S. lavandulifolia

Phenolic content** (TAE/g extract)

Flavonoid contentn.s (Rutin equivalents/g extract)

FRAP (EC1)* (mg/mL)

DPPH (IC50)* (mg/mL)

TEAC-I (IC50)* (mg/mL)

TEAC-II (IC50)* (mg/mL)

TEAC-III (IC50)* (mg/mL)

208 – 0.20a,{ 103 – 1.78b 180 – 2.98b 99 – 3.34a

10.46 – 0.17b 10.05 – 0.13b 22.18 – 0.22a 9.05 – 0.45b

0.81 – 0.056b 3.76 – 0.094a 2.62 – 0.087a 2.98 – 0.095a

1.56 – 0.78b 3.05 – 1.11a 1.41 – 0.16b 2.32 – 1.27ab

1.78 – 0.13b 5.01 – 0.10a 3.61 – 0.15ab 5.02 – 0.096a

1.47 – 0.10b 3.13 – 0.15a 2.44 – 0.12ab 4.08 – 0.18a

1.21 – 0.10b 2.51 – 0.08ab 2.11 – 0.07ab 3.77 – 0.09a

n.s Not significant. *Significant at P £ .05. **Significant at P £ .01. { Means with different superscript letters in a column are statistically significant at 5% level probability. DPPH, 1,1-diphenyl-2-picrylhydrazyl; FRAP, ferric reducing/antioxidant power; TEAC, trolox equivalent antioxidant capacity.

is expressed as RE/g dry weight extract. No significant difference (P > .05) in the flavonoid content was observed between the plants extracts (Table 2). The total flavonoid content of the extracts varied considerably from 10.05 to 22.18 RE/g extract (Table 2). The total flavonoid content of the various extracts was D. multicaule (22.18 RE/g extract) > T. daenensis (10.46 RE/g extract) > S. bachtiarica (10.05 RE/g extract) > S. lavandulifolia (9.05 RE/g extract). Our results agree with those of earlier studies by Gu et al.28 and Gohari et al.29 that determined the flavonoid contents in Dracocephalum moldavica and Dracocephalum kotschyi. A study by Dastmalchi et al.30 showed that the phenolic and flavonoid contents (quantified using luteolin-7-O-glucoside) of water extract of Dracocephalum moldavica were 145.3 mg of GAE/g extract and 26.01 mg/g dry weight extract, respectively. Nickavar and Esbati.31 have reported that the total phenolic and flavonoid contents of ethanolic extracts for three Thymus species ranged from 295.57 to 337 lg rutin/mg extract and 50.39 to 35.21 lg rutin/mg extract, respectively. The total phenolic and flavonoid contents of the extract of T. daenensis were 295 and 5.21 lg/mg extract, respectively.31 Khanavi et al.32 reported that the phenolic content of nine extract species of Stachys ranged from 4.30 (Stachys trinervis) to 32.94 (Stachys persica) mg of GAE/g extract. In a report by Molan et al.33 the phenolic content of hydrolic and ethanolic extracts of S. lavandulifolia collected from Iraq was 26.5 and 29.2 mg of GAE/g DW, respectively. In addition, a study by Bouayed et al.34 with S. lavandulifolia flowers showed that the phenolic and flavonoid contents were 14.1 mg of GAE/g DW and 4.02 mg CE/g DW. Goli et al.35 reported that the phenolic content of seed oil of D. kotschyi was 96.36 mg gallic acid/kg oil. Pla´nder et al.36 have reported that the phenolic and flavonoid contents in various extracts from the aerial parts of Satureja hortensis ranged from 0.040 to 0.13 g pyrogallol equivalent/100 g extract and 1.37% to 7.09% w/w, respectively. The phenolic content of essential oils of Satureja sahendica was 24.8 to 25.6 of GAE/g essential oil.37 The differences between current results and the previous report may be attributed to the differences in the sources of the samples and the extraction method.

Ferric reducing/antioxidant power The FRAP assay was used to measure the antioxidant effect as reducing ability. The antioxidant potential of four extracts was estimated from the ability to reduce TPTZ-Fe (III) to TPTZ-Fe (II) complex.21 The antioxidant activity was expressed as EC1. Significant differences (P £ .05) in EC1 values were observed among the samples. The EC1 values were 0.08, 0.03, 0.11, and 0.09 at 10 mg/mL for D. multicaule, T. daenensis, S. bachtiarica, and S. lavandulifolia extracts, respectively (Table 2). The findings of the present study are similar to Agbor et al.38 who indicated that the methanol extracts of Thymus vulgaris leaves and Irvingia gabonensis seeds had the highest antioxidant capacity in FRAP free antioxidant compared with 12 spice/herb extracts, and T. vulgaris ranked second in free polyphenol content. In our study, T. daenensis showed the highest antioxidant capacity in FRAP. ABTS radical scavenging activity The ABTS radical scavenging test widely is used to determine the antioxidant activity of both hydrophilic and lipophilic compounds.23 The ABTS assay also is an excellent tool for determining the antioxidant activity of hydrogendonating and chain-breaking antioxidants.39 Results indicated that there was a significant difference (P £ .05) between the extracts for IC50 values. The lowest IC50 values were obtained from the extract of T. daenensis with 1.78, 1.41, and 1.21 mg/mL at 2, 4, and 6 min, respectively (Table 2). Nickavar and Esbati31 reported that the scavenging activity of the extract from Thymus pubescens was higher than the extracts from Thymus daenensis and Thymus kotschyanus. The results of a study34 indicated that the antioxidant activity quantified by a vitamin C equivalent (VCE) for the extract of S. lavandulifolia was 15.4 mg VCE/g DW. DPPH radical scavenging activity (1,1-diphenyl-2-picrilhydrazyl) The DPPH is a stable free radical that is widely accepted as a tool for estimating the free radical scavenging activities

1155

BIOACTIVITY OF SOME OF IRANIAN HERBS (LAMIACEAE)

FIG. 2. Inhibition of DPPH by methanol extracts from T. daenensis (a), S. lavandulifolia (b), D. multicaule (c), and S. bachtiarica (d). DPPH, 1,1-diphenyl-2-picrylhydrazyl. *, Observation data.

of antioxidants.40 Antioxidant molecules can quench DPPH radicals (by providing a hydrogen atom or electron donation) and convert them to a colorless product.22 The lower IC50 value indicates a stronger ability of the extract to act as a DPPH scavenger, while the higher IC50 value indicates a lower scavenging activity of the scavengers as more scavengers were required to achieve a 50% scavenging reaction. The IC50 values were found to be 35.3, 39.0, 57.8, and 76.2 mg/mL for D. multicaule, T. daenensis, S. lavandulifolia, and S. bachtiarica, respectively (Table 2). The D. multicaule extract demonstrated the most potent activity (P £ .05). The results, however, showed no significant difference between activities of the extracts of D. multicaule and T. daenensis (P > .05). The extracts of S. lavandulifolia and S. bachtiarica revealed moderate scavenging activities (Fig. 2).

Correlation between antioxidant capacity and phenolic and flavonoid contents The correlation coefficients between the antioxidant activity and total phenolic and flavonoid contents of the medicinal plants were calculated (Table 3). The total phenolic content, as measured by the Folin–Ciocalteu reagent, correlated significantly with the FRAP and TEAC assays (r = 0.95 and 0.99, respectively). The results suggested that the phenolic compounds contributed significantly to the antioxidant capacity of the medicinal herbs. However, no significant correlation was observed between the DPPH, TEAC, and FRAP assays with flavonoid content in the extracts. The findings of the present study result in a good agreement with Andarwulan et al.,41 which suggested that the flavonoid content was not correlated with antioxidant activity

Table 3. Correlation Matrix Showing Relationship Between Antioxidant Indices (FRAP, DPPH, and TEAC), Total Phenols, and Flavonoid Content from Four Herbs (Lamiaceae)

Extract yield Total phenolic content Flavonoid content FRAP assay DPPH assay TEAC-T1 TEAC-T2 TEAC-T3

EY

TPC

FC

FRAP

DPPH

TEAC-T1

TEAC-T2

TEAC-T3

1 - 0.942* - 0.341 0.797 0.666 0.936* 0.998** 0.981**

1 0.183 - 0.953** - 0.755 - 9.998** - 0.957** - 0.880*

1 - 0.003 - 0.613 - 0.175 - 0.291 - 0.272

1 0.745 0.958* 0.791 0.705

1 0.758 0.621 0.514

1 0.931* 0.872

1 0.989**

1

*Significant at P £ .05. **Significant at P £ .01. EY, extract yield (%); TPC, total phenolic content (TAE/g extract); FC, flavonoid content (rutin equivalents/g extract); FRAP (EC1) (mg/mL); DPPH (IC50) (mg/ mL); TEAC-T1 (IC50) at 2 min (mg/mL); TEAC-T2 (IC50) at 4 min (mg/mL); TEAC-T3 (IC50) at 6 min (mg/mL).

1156

GHASEMI PIRBALOUTI ET AL.

in the DPHH, ABTS, and reducing power assays. The antioxidant activity of the total phenolic and flavonoid contents depends highly on the molecular structure, suggesting that the antioxidant activity may be determined by individual polyphenols and/or flavonoids and not the total content. Moreover, the DPPH method indicated no significant correlation between the total phenolic and flavonoid content and antioxidant activity (Table 3). Other investigators13,41–47 have observed a linear response between the total phenolic and antioxidant capacity in FRAP. Several investigations48,49 have studied correlations between bioactive compounds and antioxidant activity by the DPHH assay in numerous herbs and spices. Little information, however, is known concerning the total phenolic and antioxidant activity by the TEAC assay. In this study, the TEAC antioxidant activity values increased with increasing polyphenol content of the plant extracts analyzed, in agreement with previous studies.48,49 Literature review shows the presence of the two main groups of secondary metabolites in the genus of Thymus and Satureja, volatile terpenoids, and polyphenolic compounds.50,51 The volatile terpenoids and polyphenolic compounds are mainly responsible for the biological effects of the genus, particularly, antioxidant activities.51 Thymol and carvacrol belonging to monoterpenoid phenol are the main constitutes in the essentials oils from T. daenensis and S. bachtiarica.50,51 On the other hand, the presence of various polyphenolic constituents, especially flavonoids and phenolic acids, is well known in Thymus and Satureja plants. In conclusion, the present study is apparently the first report of quantitative flavonoid and phenol profiles for three Iranian medicinal plants, including D. multicaule, T. daenensis, and S. bachtiarica. These medicinal herbs are ordinarily used for pharmaceutical purposes and also as health foods and teas in Iran. The results of the current study demonstrated the antioxidant activities by three assays for the methanol extracts of thyme, savory, dragonhead, and woundwort. A positive correlation was obtained between the total phenolic content and antioxidant activity in the FRAP and TEAC assays. A main finding in this study was that the medicinal herbs demonstrated a much higher antioxidant activity and contained significantly more phenolics and flavonoids than other Iranian species of herbs that are considered as good natural sources of dietary antioxidants. This study showed that T. daenensis possess a significant reducing power and free radical scavenging ability in vitro. In total, the significant antioxidant activity of the extracts of the studied herbs, especially T. daenensis, provides a scientific validation for the traditional use of the plant as an accessible source of natural antioxidants with consequent health benefits. Further work on the isolation and identification of active compounds and their efficacy needs to be done. In addition, the antioxidative properties in vivo of T. daenensis and other studied herbs should be the objective of future research.

ACKNOWLEDGMENT This work was supported by the School of Pharmacy, Ahwaz Jondishapur University of Medical Sciences, Ahwaz, Iran.

AUTHOR DISCLOSURE STATEMENT No competing financial interests exist. REFERENCES 1. Cai Y, Luo Q, Sun M, Corke H: Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 2004;74:2157–2184. 2. Halliwell B: Free radicals, antioxidants, and human disease: curiosity, cause, or consequence? Lancet 1994;344:721–724. 3. Niki E: Free radicals, antioxidants, and cancer. In: Food Factors for Cancer Prevention (Ohigashi H, Osawa T, Terao J, Watanabe S, Yoshikawa T, eds.). Springer, Tokyo, 1997, pp. 55–57. 4. Poulson HE, Prieme H, Loft S: Role of oxidative DNA damage in cancer initiation and promotion. Eur J Cancer Prev 1998;7:9–16. 5. Yanishlieva NV, Marinova EY, Pokorny J. Natural antioxidants from herbs and spices. Eur J Lipid Sci Technol 2006;108: 776–793. 6. Larson RA: The antioxidants of higher plants. Phytochemistry 1988;27:969–978. 7. Shahidi F, Naczk M: Food Phenolics: Sources, Chemistry, Effects and Applications. Technomic Publishing Company, Basel, Switzerland, 1995. 8. Cotelle N, Bernier JL, Catteau JP, Pommery J, Wallet JC, Gaydou EM: Antioxidant properties of hydroxyflavones. Free Radic Biol Med 1996;20:35–43. 9. Velioglu YS, Mazza G, Gao L, Oomah BD: Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. J Agric Food Chem 1998;46:4113–4117. 10. Cai YZ, Sun M, Corke H: Antioxidant activity of betalains from plants of the Amaranthaceae. J Agric Food Chem 2003;51:2288– 2294. 11. Zheng W, Wang SY: Antioxidant activity and phenolic compounds in selected herbs. J Agric Food Chem 2001;49:5165–5170. 12. Conforti F, Menichini F, Formisano C, Rigano D, Senatore F, Arnold NA, Piozzi F: Comparative chemical composition, free radical-scavenging and cytotoxic properties of essential oils of six Stachys species from different regions of the Mediterranean area. Food Chem 2009;116:898–905. 13. Ghasemi Pirbalouti A, Hashemi M, Ghahfarokhi FT: Essential oil and chemical compositions of wild and cultivated Thymus daenensis Celak and Thymus vulgaris L. Ind Crops Prod 2013; 48:43–48. 14. Mozaffarian V: A Pictorial Dictionary of Botany (Botanical Taxonomy): Latin - English - French - German - Persian (Mozaffarian V, ed.). Farahang Moaser, Tehran, 2008, p. 522. 15. Zargari A: Iranian Medicinal Plants, Vol. 1–6. University Publication, Tehran, Iran, 1982–1992. 16. Ghasemi Pirbalouti A: Medicinal plants used in Chaharmahal and Bakhtyari districts, Iran. Herba Polon 2009;55:69–75. 17. Ghasemi Pirbalouti A, Malekpoor F, Hamedi B: Ethnobotany and antimicrobial activity of medicinal plants of Bakhtiari Zagross mountains Iran. J Med Plant Res 2012;6:675–679. 18. Ghasemi Pirbalouti A, Mohammadi M: Phytochemical composition of the essential oil of different populations of Stachys lavandulifolia Vahl. Asian Pac J Trop Biomed 2013;3:123–128. 19. Singleton VL, Rossi JA: Colorimetry of total phenolics with phosphomolybdic–phosphotungstic acid reagents. Am J Enol Vitic 1965;16:144–158. 20. Quettier-Deleu C, Gressier B, Vasseur J, et al.: Phenolic compounds and antioxidant activities of buckwheat (Fagopyrum

BIOACTIVITY OF SOME OF IRANIAN HERBS (LAMIACEAE)

21.

22.

23.

24.

25.

26.

27.

28.

29.

30.

31.

32.

33.

34.

35.

36.

37.

esculentum Moench) hulls and flour. J Ethnopharmacol 2000;72: 35–42. Benzie IIF, Strain JJ: The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: The FRAP Assay. Anal Biochem 1996;239:70–76. Brand-Williams W, Cuvelier ME, Berset C: Use of a free radical method to evaluate antioxidant activity. Lebensm Wiss Technol 1995;28:25–30. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, RiceEvans C: Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 1999; 26:1231–1237. Rabbani M, Sajjadi SE, Jalali A: Hydroalcoholic extract and fractions of Stachys lavandulifolia Vahl: effects on spontaneous motor activity and elevated plus-maze behavior. Phytother Res 2005;19:854–858. Yogesh K, Jha SN, Ahmad T: Antioxidant potential of aqueous extract of some food grain powder in meat model system. J Food Sci Technol Mysore 2012;1–6. Xu BJ, Chang SKC: A comparative study on phenolic profiles and antioxidant activities of legumes as affected by extraction solvents. J Food Sci 2007;72:S159–S166. Nithiyanantham S, Selvakumar S, Siddhuraju P: Total phenolic content and antioxidant activity of two different solvent extracts from raw and processed legumes, Cicer arietinum L and Pisum sativum L. J Food Comp Anal 2012;27:52–60. Gu HF, Chen RY, Sun YH, Lui F: Studies on chemical constituents from herb of Dracocephalum moldavica. Zhongguo Zhong Yao Za Zhi 2004:29:232–234. Gohari AR, Saeidnia S, Matsuo K, et al.: Flavonoid constituents of Dracocephalum kotschyi growing in Iran and their trypanocidal activity. Nat Med 2003;57:250–252. Dastmalchi H, Dorman D, Kosarb M, Hilttunen R: Chemical composition and in vitro antioxidant evaluation of a water soluble Moldavican balm (Dracocephalummoldavica L.) extract. LWT Food Sci Technol 2007;40:1655–1663. Nickavar B, Esbati N: Evaluation of the antioxidant capacity and phenolic content of three Thymus species. J Acupunct Meridian Stud 2012;5:119–125. Khanavi K, Saghari Z, Mohammadirad A, Khademi R, Hadjiakhoondi A, Abdollahi M: Comparison of antioxidant activity and total phenols of some date varieties. Daru J Pharm Sci 2009;17:104–108. Molan AL, Faraj AM, Mahdy AS: Antioxidant activity and phenolic content of some medicinal plants traditionally used in Northern Iraq. Phytopharmacology 2012;2:224–233. Bouayed J, Piri K, Rammal H, et al.: Comparative evaluation of the antioxidant potential of some Iranian medicinal plants. Food Chem 2007;104:364–368. Goli SAH, Sahafi SM, Rashidi B, Rahimmalek M: Novel oilseed of Dracocephalum kotschyi with high n-3 to n-6 polyunsaturated fatty acid ratio. Ind Crop Prod 2013;43:188–193. Pla´nder S, Gontaru L, Blazics B, et al.: Major antioxidant constituents from Satureja hortensis L. extracts obtained with different solvents. Eur J Lipid Sci Technol 2012;114:772–779. Sadeghi Ghotbabadi F, Alizadeh A, Zadehbagheri M, Kamelmanesh MM, Shaabani M: Phytochemical composition of the

38.

39. 40.

41.

42.

43.

44.

45.

46.

47.

48.

49.

50.

51.

1157

essential oil, total phenolic content, antioxidant and antimicrobial activity in Iranian Satureja sahendica Bornm at different ontogenesis conditions. J Med Plant Res 2012;6:3525–3534. Agbor GA, Julius E, Ngogang JY, Xinxing C, Vinson JA: Antioxidant capacity of some herbs/spices from Cameroon: a comparative study of two methods. J Agric Food Chem 2005;53: 6819–6824. Leong LP, Shui G: An investigation of antioxidant capacity of fruits in Singapore markets. Food Chem 2002;76:69–75. Hu FL, Lu RL, Huang B, Ming L: Free radical scavenging activity of extracts prepared from fresh leaves of selected Chinese medicinal plants. Fitoterapia 2004;75:14–23. Andarwulan N, Batari R, Sandrasari DA, Bolling B, Wijaya H: Flavonoid content and antioxidant activity of vegetables from Indonesia. Food Chem 2010;121:1231–1235. Benzie IF, Strain JJ: Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol 1999;299:15–27. Luximon-Ramma A, Bahorun T, Soobrattee MA, Aruoma OI: Antioxidant activities of phenolic, proanthocyanidin, and flavonoid components in extracts of Cassia fistula. J Agric Food Chem 2002;18:5042–5047. Thaipong K, Boonprakob U, Crosby K, Cisneros-Zevallos L, Bryne DH: Comparison of ABTS, DPPH, FRAP and ORAC assays for estimating antioxidant activity from guava fruit extracts. J Food Comp Anal 2006;19:669–675. Chizzola R, Michitsch H, Franz C: Antioxidative properties of Thymus vulgaris leaves: comparison of different extracts and essential oil chemotypes. J Agric Food Chem 2008;27:6897–6904. Dudonne S, Vitrac X, Coutiere P, Woillez M, Merillon JM: Comparative study of antioxidant properties and total phenolic content of 30 plant extracts of industrial interest using DPPH, ABTS, FRAP, SOD, and ORAC Assays. J Agric Food Chem 2009; 57:1768–1774. Asadi S, Ahmadiani A, Esmaeili MA, Sonboli A, Ansari N, Khodagholi F: In vitro antioxidant activities and an investigation of neuroprotection by six Salvia species from Iran: a comparative study. Food Chem Toxicol 2010;48:1341–1349. Corral-Aguayo RD, Yahia EM, Carrillo-Lopez A, GonzalezAguilar G: Correlation between some nutritional components and the total antioxidant capacity measured with six different assays in eight horticultural crops. J Agric Food Chem 2008;56:10498– 10504. Proteggente AR, Pannala AS, Paganga G, et al.: The antioxidant activity of regularly consumed fruit and vegetables reflects their phenolic and vitamin C composition. Free Radic Res 2002;36: 217–233. Ghasemi Pirbalouti A, Hashemi M, Ghahfarokhi FT: Essential oil and chemical compositions of wild and cultivated Thymus daenensis Celak and Thymus vulgaris L. Ind Crop Prod 2013;48: 43–48. Ghasemi Pirbalouti A, Dadfar S: Chemical constituents and antibacterial activity of essential oil of Satureja bachtiarica (Lamiaceae). Acta Pol Pharm 2013;70:933–938.

Antioxidant activity, total phenolic and flavonoid contents of some medicinal and aromatic plants used as herbal teas and condiments in Iran.

Total phenolic and flavonoid contents and antioxidant activity of four Iranian herbs (Lamiaceae) were investigated. The antioxidant activity of methan...
446KB Sizes 0 Downloads 9 Views