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Antioxidant activities in Selected Herb

Antioxidant activities in Selected Herb

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Subscriber access provided by DigiTop | USDA's Digital Desktop Library
Journal of Agricultural and Food Chemistry is published by the American ChemicalSociety. 1155 Sixteenth Street N.W., Washington, DC 20036
Article
Antioxidant Activity and Phenolic Compounds in Selected Herbs
Wei Zheng, and Shiow Y. Wang
J. Agric. Food Chem.
,
2001
, 49 (11), 5165-5170 • DOI: 10.1021/jf010697n
Downloaded from http://pubs.acs.org on January 13, 2009
More About This Article
Additional resources and features associated with this article are available within the HTML version:Supporting InformationLinks to the 27 articles that cite this article, as of the time of this article downloadAccess to high resolution figuresLinks to articles and content related to this articleCopyright permission to reproduce figures and/or text from this article
 
Antioxidant Activity and Phenolic Compounds in Selected Herbs
Wei Zheng
and Shiow Y. Wang*
Fruit Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service,U. S. Department of Agriculture, Beltsville, Maryland 20705
The antioxidant capacities (oxygen radical absorbance capacity, ORAC) and total phenolic contentsin extracts of 27 culinary herbs and 12 medicinal herbs were determined. The ORAC values andtotal phenolic contents for the medicinal herbs ranged from 1.88 to 22.30
µ
mol of Trolox equivalents(TE)/g of fresh weight and 0.23 to 2.85 mg of gallic acid equivalents (GAE)/g of fresh weight,respectively.
Origanum
×
majoricum
,
O. vulgare
ssp
. hirtum
, and
Poliomintha longiflora
have higherORAC and phenolic contents as compared to other culinary herbs. The ORAC values and totalphenolic content for the culinary herbs ranged from 2.35 to 92.18
µ
mol of TE/g of fresh weight and0.26 to 17.51 mg of GAE/g of fresh weight, respectively. These also were much higher than valuesfound in the medicinal herbs. The medicinal herbs with the highest ORAC values were
Catharanthusroseus
,
Thymus vulgaris
,
Hypericum perforatum
, and
Artemisia annua
. A linear relationship existedbetween ORAC values and total phenolic contents of the medicinal herbs (
 R
)
0.919) and culinaryherbs (
 R
)
0.986). High-performance liquid chromatography (HPLC) coupled with diode-arraydetection was used to identify and quantify the phenolic compounds in selected herbs. Among theidentified phenolic compounds, rosmarinic acid was the predominant phenolic compound in
Salviaofficinalis
,
Thymus vulgaris, Origanum
×
majoricum
, and
P. longiflora
, whereas quercetin-3-
O
-rhamnosyl-(1
f
2)-rhamnosyl-(1
f
6)-glucoside and kaempferol-3-
O
-rhamnosyl-(1
f
2)-rhamnosyl-(1
f
6)-glucoside were predominant phenolic compounds in
Ginkgo biloba
leaves.
Keywords:
Antioxidant; phenolics; medicinal herbs; culinary herbs
INTRODUCTION
Antioxidants are compounds that can delay or inhibitthe oxidation of lipids or other molecules by inhibitingthe initiation or propagation of oxidizing chain reactions(
1
). The antioxidant activity of phenolic compounds ismainly due to their redox properties, which can playan important role in adsorbing and neutralizing freeradicals, quenching singlet and triplet oxygen, or de-composing peroxides (
2
). In general, there are two basiccategories of antioxidants, natural and synthetic. Re-cently, interest has increased considerably in findingnaturally occurring antioxidants for use in foods ormedicinal materials to replace synthetic antioxidants,which are being restricted due to their carcinogenicity(
3
). Herbs have been used for a large range of purposesincluding medicine, nutrition, flavorings, beverages,dyeing, repellents, fragrances, cosmetics, charms, smok-ing, and industrial uses. Since prehistoric times, herbswere the basis for nearly all medicinal therapy untilsynthetic drugs were developed in the nineteenthcentury. Today, herbs are still found in 40% of prescrip-tion drugs (
4
). Culinary herbs have been grown and usedfor hundreds of years, and they are becoming increas-ingly popular in the United States for their ability toenhance and complement the flavors of a wide varietyof foods (
5
).Even though a variety of herbs are known to besources of phenolic compounds, their compositional dataare insufficient (
). Moreover, various herbs along withvegetables and fruits contain numerous phytochemicalsin addition to phenolic compounds, such as nitrogencompounds, carotenoids, and ascorbic acid (
1
,
). Manyof these phytochemicals possess significant antioxidantcapacities that are associated with lower incidence andlower mortality rates of cancer in several human cohort(
1
).The purpose of this study was to (i) evaluate a varietyof culinary and medicinal herbs that were growing inthe same location and same conditions with respect totheir total phenolic content and antioxidant activity tofind new potential sources of natural antioxidants; (ii)evaluate the relationship between phenolic content andantioxidant activity; and (iii) develop chromatographicprocedures to identify and quantify phenolic antioxi-dants in selected herbs by high-performance liquidchromatography (HPLC).
MATERIALS AND METHODS
Chemicals.
2
,2
-Azobis(2-amidinopropane) dihydrochloride(AAPH) was purchased from Wako Chemicals USA Inc,(Richmond VA). (
 R
)-Phycoerythrin (R-PE) was obtained fromSigma (St. Louis, MO). 6-Hydroxy-2,3,7,8-tetramethylchro-man-2-carboxylic acid (Trolox) was purchased from Aldrich(Milwaukee, WI). Acetonitrile, methanol, acetone, and waterwere of HPLC grade and were purchased from Baxter (Muskeg-on, MI). Authentic standards were obtained from SigmaChemical Co. (St. Louis, MO), Fisher Scientific Co. (Pittsburgh,PA), and Indofine Chemical Co. (Somerville, NJ).
Sample Preparation.
The 39 different herbs (Table 1),including medicinal and culinary herbs, were collected onSeptember 2000 from the National Herb Garden, which is part* To whom correspondence should be addressed[telephone (301) 504-5776; fax (301) 504-5062; E-mail wangs@ba.ars.usda.gov].
Visiting scientist from Institute of Environmental Science,Zhejiang University at Yuquan, Hangzhou, Zhejiang 310027,P. R. China.
5165
J. Agric. Food Chem.
2001,
49,
5165
517010.1021/jf010697n This article not subject to U.S. Copyright. Published 2001 by the American Chemical SocietyPublished on Web 09/28/2001
 
of the U. S. National Arboretum in Washington, D.C. All theherbs were grown in the same location and same conditionsto avoid variations of oxygen radical absorbance capacity(ORAC) values due to environmental factors. All samples werestored in a freezer at
-
80 °C before analysis. Herbs (2.0 g)were extracted with 15 mL of phosphate buffer (75 mM, pH7.0) using a Polytron homogenizer (Brinkmann Instruments,Inc., Westbury, NY) for 1 min and were then centrifuged at20000
g
for 20 min. The supernatant was recovered and usedfor the ORAC and total phenolic compound assay after suitabledilution with phosphate buffer (75 mM, pH 7.0).
Oxygen Radical Absorbance Capacity (ORAC) Assay.
The procedure for performing ORAC assays for the herbsamples were based on a previous report of Wang and Lin (
8
),which was modified from a method described by Cao et al. (
9
).This assay measures the ability of antioxidant compounds intest materials to inhibit the decline of R-PE fluorescence thatis induced by a peroxyl radical generator, AAPH. The reactionmixture contained 1.7 mL of 75 mM phosphate buffer (pH 7.0),100
µ
L of R-PE (3.4 mg/L), 100
µ
L of 320 nM AAPH, and 100
 µ
L of sample. Trolox, a water-soluble analogue of vitamin E,was used as a control antioxidant standard. The fluorescenceof R-PE was determined and recorded every 5 min at theexcitation wavelength of 540 nm and emission wavelength of 570 nm using a Shimadzu RF-Mini 150 recording fluorometer(Columbia, MD) until the fluorescence of the last readingdeclined to
<
5% of the first reading. The final results (ORACvalue) were calculated using the differences of areas underthe quenching curves of R-PE between a blank and a sampleand expressed as micromoles of Trolox equivalents (TE) pergram of fresh weight.
Total Phenolic Compound Analysis.
The amount of totalphenolics in the herb extracts was determined with the Folin-Ciocalteu reagent according to the method of Slinkard andSingleton (
10
) using gallic acid as a standard. Samples (200
 µ
L, two replicates) were introduced into test cuvettes, and then1.0 mL of Folin-Ciocalteu’s reagent and 0.8 mL of Na
2
CO
3
(7.5%) were added. The absorbance of all samples was mea-sured at 765 nm using the Shimadzu UV
-
Vis spectrophotom-eter after incubating at 30 °C for 1.5 h. Results were expressedas milligrams of gallic acid equivalent (GAE) per gram of freshweight.
HPLC Analysis of Selected Herbs.
The sample (2.0 g)was extracted twice with 15 mL of acetone using a Polytronhomogenizer (Brinkmann Instruments, Inc., Westbury, NY)for 1 min. The extract was centrifuged, and the residue waswashed and agitated twice with 5 mL of solvent. The combinedextract was evaporated to dryness under reduced pressure.The residue was dissolved in 4 mL of methanol, and 20
µ
Laliquots were analyzed by HPLC. The herb extracts used forHPLC analysis were passed through a 0.45-
 µ
m filter (Milli-pore, MSI, Westboro, MA) before injection into a reverse phaseNOVA-PAK C
18
column (150
×
39 mm i.d., particle size 4
µ
m)with a guard column (NOVA-PAK C
18
, 20
×
3.9 mm i.d.,particle size 4
µ
m) at ambient temperature (20 °C). A Waters600E system controller coupled with a photodiode arraydetector (Waters 990 series) was used. The mobile phase wasacetonitrile (A) and acidified water containing 2.5% formic acid
Table 1. Total Phenolic Content and Antioxidant Activity (ORAC) in Various Herbal Extracts
 a
common name botanical name typetotal phenolic
b
(mg of GAE/g of fresh weight)ORAC
c
(
 µ
mol of TE/ g of fresh weight)balsam pear
Momordica charantia
medicinal 0.43
(
0.08 3.43
(
0.11creeping thyme
Thymus praecox
ssp.
arcticus
medicinal 1.81
(
0.04 13.40
(
0.12feverfew
Tanacetum parthenium
medicinal 0.87
(
0.06 10.07
(
0.15garden sage
Salvia officinalis
medicinal 1.34
(
0.09 13.28
(
0.40garden thyme
Thymus vulgaris
medicinal 2.13
(
0.11 19.49
(
0.21Madagascar periwinkle
Catharanthus roseus
medicinal 2.85
(
0.11 22.30
(
0.54maidenhair tree
Ginkgo biloba
medicinal 1.57
(
0.05 13.18
(
0.24peppermint
Mentha
×
 piperita
medicinal 2.26
(
0.16 15.84
(
0.42Saint John’s wort
Hypericum perforatum
medicinal 2.78
(
0.12 16.77
(
0.22sweet wormwood
Artemisia annua
medicinal 1.54
(
0.06 15.69
(
0.37true aloe
Aloe vera
medicinal 0.23
(
0.00 1.88
(
0.05valerian
Valerian officinalis
medicinal 1.78
(
0.12 15.82
(
0.61caraway
Carum carvi
culinary 1.05
(
0.00 10.65
(
0.29chives
Allium schoenoprasum
culinary 1.05
(
0.05 9.15
(
0.28cuban oregano
Plectranthus amboinicus
culinary 0.34
(
0.00 4.71
(
0.14dandelion
Taraxacum officinale
culinary 0.26
(
0.02 2.35
(
0.14dill
Anethum graveolens
culinary 3.12
(
0.06 29.12
(
0.29English lavender
Lavandula angustifolia
culinary 1.50
(
0.13 16.20
(
0.11fennel
Foeniculum vulgare
culinary 0.68
(
0.00 5.88
(
0.09Greek mountain oregano
Origanum vulgare
ssp.
hirtum
culinary 11.80
(
0.60 64.71
(
1.05hard sweet marjoram
Origanum
×
majoricum
culinary 11.65
(
0.29 71.64
(
1.25lemon balm
Melissa officinalis
culinary 1.26
(
0.04 9.54
(
0.23lemon thyme
Thymus
×
citriodorus
culinary 1.78
(
0.03 13.28
(
0.33lemon verbena
Aloysia triphylla
culinary 1.55
(
0.10 17.38
(
0.35lovage
Levisticum officinale
culinary 2.63
(
0.05 21.54
(
0.35Mexican oregano
Poliomintha longiflora
culinary 17.51
(
0.22 92.18
(
0.72orange mint
Mentha aquatica
culinary 2.26
(
0.10 19.80
(
0.43parsley
Petroselinum crispum
culinary 1.12
(
0.01 11.03
(
0.13pineapple sage
Salvia elegans
culinary 1.31
(
0.08 11.55
(
0.42purple amaranth
Amaranthus cruentus
culinary 3.41
(
0.11 28.92
(
0.21rose geranium
Pelargonium graveolens
culinary 7.34
(
0.36 38.75
(
0.61rosemary
Rosmarinus officinalis
culinary 2.19
(
0.15 19.15
(
0.63salad burnet
Sanguisorba minor 
culinary 0.99
(
0.07 8.33
(
0.13society garlic
Tulbaghia violacea
culinary 1.03
(
0.10 7.50
(
0.60spearmint
Mentha spicata
culinary 0.94
(
0.15 8.10
(
0.26sweet basil
Ocimum basilicum
culinary 2.23
(
0.15 14.27
(
0.45sweet bay
Laurus nobilis
culinary 4.02
(
0.90 31.70
(
0.97Vietnamese coriander
Polygonum odoratum
culinary 3.09
(
0.12 22.30
(
0.68winter savory
Satureja montana
culinary 3.16
(
0.02 26.34
(
0.17LSD
0.05
0.27 0.79
a
Data expressed as mean
(
SEM.
b
Data expressed as milligrams of gallic acid (GAE) equivalents per gram of fresh weight.
c
Dataexpressed as micromoles of Trolox equivalents per gram of fresh weight.
5166
J. Agric. Food Chem.,
Vol. 49, No. 11, 2001 Zheng and Wang

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