Industrial Crops and Products 33 (2011) 382–388

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Industrial Crops and Products

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Phenolic composition and antioxidant capacity of six artemisia species

Isabel S. Carvalho a,∗ , Teresa Cavaco a , Maria Brodelius b
a
IBB/CGB – Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, Ed. 8, 8005-139 Faro, Portugal
b
School of Pure and Applied Natural Sciences, University of Kalmar, SE-39182 Kalmar, Sweden

a r t i c l e i n f o a b s t r a c t

Article history: The aim of this work was to establish the antioxidant capacities and the polyphenolic profile of six dif-
Received 9 March 2010 ferent Artemisia species that could potentially be used in the human diet. A high-performance liquid
Received in revised form 5 November 2010 chromatographic method coupled with photodiode-array detector was used to identify and quantify
Accepted 12 November 2010
individual phenolic compounds of the Artemisia leaves. A total of 18 polyphenolic compounds were iden-
Available online 28 December 2010
tified and quantified in Artemisia leaves, including hydroxybenzoic acids (4), hydroxycinnamic acids (5),
flavonols (3), and catechins (2). It was observed that total phenolic content of A. annua and A. stelle-
Keywords:
riana leaves were significantly lower than the other four species. Ferulic and caffeic conjugates acids
Antioxidant activity
Artemisia spp.
were the most dominant hydroxycinnamic acid and gallic acid and catechin were the most dominant
High-performance liquid chromatographic hydroxycinnamic acid and catechins respectively. According to DPPH assays, the antioxidant capacity of
(HPLC) A. arborescens spp., A. ludoviciana spp., A. oleandica spp. and A. princepts spp., were found to be higher
Flavonoids (reflecting a 2-fold difference) than that of the other two species. Compared with those of major com-
Phenolic compounds mercial leafy vegetables, leaves of Artemisia contain a higher content of flavonoids and phenolic acids,
which provide significant health benefits and may be used as natural colorants.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction lar disease, cancer, atherosclerosis, and other age-related diseases

In recent years, there has been a global trend toward the use
of natural phytochemicals present in natural resources – such as
vegetables, fruits, oilseeds and herbs – as antioxidants and func-
tional ingredients (Elliott, 1999; Kaur and Kapoor, 2001; Namiki,
1990). Polyphenolic compounds are a large group of secondary
metabolites widely distributed in plants, which can be divided into
two major subgroups: flavonoids and phenolic acids. Flavonoids,
perhaps the most important single group of phenolics in foods,
comprise a group of over 4000 aromatic plant compounds, which
include anthocyanins, proanthocyanidins, flavonols and catechins.
On the other hand, phenolic acids include hydroxycinnamic acids
(e.g. caffeic or ferrulic acid conjugates, sinapic acid) and hydrox-
ybenzoic acids (e.g. benzoic, gentisic or p-anisic acids) (Carvalho
et al., 2010). Epidemiological and intervention studies have pro-
vided evidence of beneficial health effects of dietary fruits and
vegetables, and the beneficial effects have been attributed at least
in part to secondary metabolites, including flavonoids and hydrox-
ycinnamic acids (Nijveldt et al., 2001). Effects of selected flavonoids
in reducing the risk of various diseases including cardiovascu-
∗ Corresponding author at: Food Science Laboratory, FCT, Dept. of Biological Science
and Bioengineering, University of Algarve, Campus de Gambelas, Ed. 8, P-8005-139
Faro, Portugal. Tel.: +351 289800040; fax: +351 289811419.
E-mail address: icarva@ualg.pt (I.S. Carvalho).
have been demonstrated (Erlejman et al., 2006; Hodgson and Croft,
2006) and it is thought that these phytochemicals may provide
health benefits as antioxidants.
The genus Artemisia, small herbs and shrubs, is one of the
largest and most widely distributed genera of the Asteraceae fam-
ily. Artemisia species, widespread in nature, are frequently utilized
for the treatment of diseases such as malaria, hepatitis, cancer,
inflammation, and infections by fungi, bacteria, and viruses. This
genus is receiving growing attention presumably due to: (i) the
diversified biology and chemistry of the constituents, (ii) the fre-
quent application in traditional medical practice, and (iii) the rich
source of the plant material. They grow in temperate climates of
the Northern Hemisphere and Southern Hemisphere, usually in
dry or semi-dry habitats. Many spices of the genus Artemisia are
known as aromatic plants and have a characteristic scent or taste,
caused by monoterpenes and sesquiterpenes, which in many cases
are the reason for their application in folk medicine. Other spices
have phytotoxic activity, and are a candidate as a natural herbi-
cide (Chen et al., 1999). For instance, Artemisia arborescens spp. (A.
arborescens) is a very bitter herb indigenous in the Middle East and
is used in tea, usually with mint because it may have some hal-
lucinogenic properties; Artemisia annua L. (A. annua) is used by
Chinese herbalists in ancient times to treat fever, but has fallen
out of common use; Artemisia ludoviciana Nutt (A. ludoviciana)
is largely ignored by modern herbalists has long been a favorite
among native peoples and gypsy healers for its broad range of uses

0926-6690/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.indcrop.2010.11.005
 I.S. Carvalho et al. / Industrial Crops and Products 33 (2011) 382–388
Fig. 1. Leaves of Artemisia: (1) A. annua L., (2) A. arborescens spp., (3) A. ludoviciana Nutt., (4) A. oleandica Bess., (5) A. princeps Pamp. and (6) A. stelleriana Bess.
and dependable availability, its effects include the reproductive,
digestive, urinary and nervous systems; the leaves of Artemisia prin-
ceps Pamp. (A. princeps) have traditionally been used as teas and
foods in Japan and Artemisia stelleriana Bess. (A. stelleriana) is usu-
ally found in hind dune regions with low salinities and its medicinal
value has been linked to promote the growth hair and to stimu-
late mental faculties. So, it is well established, all over the world,
that Artemisia spp. plant greens were important foods in the tradi-
tional diet from ancient times; they consumed plants that today are
no longer generally considered for nutrition (Guil-Guerrero et al.,
1998). Some modern cultures still consume wild plants as a nor-
mal food source, obtaining fairly good amounts of several nutrients,
and it is widely accepted that leafy green vegetables are significant
nutritional sources of minerals (Kuhnlein and Paterson, 1990). In
addition, they are also used in folk medicine as anti scorbutic, anti-
spasmodic, tonic, carminative agents against bronchitis, ulcers and
as diuretics and depuratives or are used as tea and flavoring agents
in several regions (Koedam, 1986). The nutritional and medicinal
properties of those plants may be inter-linked through phytochem-
icals (Ranhotra et al., 1998). Several studies have been carried out
on edible wild plants (Guil-Guerrero et al., 1998; Özcan and Akgül,
1998). However, limited studies were carried out on polypheno-
lic contents of medicinal and aromatic Artemisia plants. Therefore,
the aim of this work was to establish the antioxidant capacities
and the polyphenolic profile of six different Artemisia species: (1)
A. annua L., (2) A. arborescens spp., (3) A. ludoviciana Nutt., (4) A. ole-
andica Bess., (5) A. princeps Pamp. and (6) A. stelleriana Bess., which
could potentially be used in the human diet. The study of polyphe-
nolic composition is an important scientific agenda for food and
nutritional sciences, which may contribute to the improvement of
conventional foods with added health benefits being very useful
383
to determine these chemicals in plants, in the field of nutrition,
pharmacology and agronomy.
2. Experimental
2.1. Chemicals
1,1-Diphenyl-2-picrylhydrazyl (DPPH), Folin–Ciocalteu
reagent, Trolox, quercetin, gallic acid, disodium fluorescein,
luteolin, chlorogenic acid, and caffeic acid were purchased from
Sigma–Aldrich Co. Ltd. (Poole, UK). Di-potassium hydrogen
orthophosphate and di-hydrogen orthophosphate di-hydrate
were purchased from Merck (Nottingham, UK), and cyanidin was
purchased from Polyphenols Laboratories (Sandnes, Norway).
2.2. Plant materials
The experiment was conducted under greenhouse conditions
(∼18–20 ◦ C) during January–April of 2007. Seeds of a cultivated
Artemisia’s (1 – A. annua, 2 – A. arborescens, 3 – A. ludoviciana, 4
– A. oleandica, 5 – A. princepts and 6 – A. stelleriana) (Fig. 1), were
sown in 1 cm × 1 cm cell size seedling trays filled with commer-
cial medium Metro 510 (O. M. Scotts, Marysville, OH). The seedling
trays were kept in the greenhouse at a temperature of ∼18–20 ◦ C,
irrigated as needed with tap water, and thinned periodically to
retain only one or two seedlings in each cell. The seedlings (21
days old) were transplanted into 500 cm3 square pots containing
the same Metro 510. Plants were fertilized three times a week with
nitrogen at 100 ␮g/mL until harvest using a 20 N–4.36 P–16.6 K
water-soluble fertilizer in the irrigation water. The aerial parts of
384 I.S. Carvalho et al. / Industrial Crops and Products 33 (2011) 382–388
Table 1
Artemisia spp. plants. Latin name, common name and family.
Latin name Common name
A. annua L. Annual Wormwood, Sweet Sagewort, Sweet Annie
A. arborescens Tree Wormwood
A. ludoviciana Nutt Moonwort, Western Mugwort
A. oleandica (Besser) Krasch. Unknown
A. princeps Pamp. Japanese Mugwort, Yomogi
A. stelleriana Bess. Beach Wormwood, Hoary Mugwort
Artemisia species were harvested at flowering stages. The collected
plant materials were dried in an oven (Selecta, Barcelona, Spain)
at 35 ◦ C for 24 h then the leaves were separated and ground in a
grinder (Moulinex, France) and stored at −20 ◦ C until further anal-
ysis. The common, scientific and family names of the plants used in
this study are given in Table 1.
2.3. Preparation of Artemisia extracts
Dried leaves of Artemisia (0.3 g) were homogenized at room
temperature with 3 mL of methanol (Merck) in a rotary mixer
(Edmund Buhler Gmg H-Ks 15, Germany) at 45 ◦ C angle and
200 rpm for 24 h and the mixture was centrifuged at 1600 × g for
10 min. The supernatant was filtered though Whatman no. 4 fil-
ter paper and kept at −20 ◦ C before analysis. The resulting extracts
were used for analysis.
2.4. Colour analyses
A direct measurement of absorbance at 420, 520, and 620 nm
in methanolic extracts was carried out using an Ultrospec 1100
pro-UV/visible spectrophotometer (GE Heathcare Life Sciences,
England). The following variables were calculated: colour inten-
sity (CI) as the sum of 420 nm, 520 nm, and 620 nm absorbance’s;
proportion of yellow (Ye %), red (Rd %) and blue colour (Bl %) were
calculated by dividing the absorbance’s at 420, 520 and 620 nm by
the colour intensity (CI), respectively (Porra et al., 1989).
2.5. Determination of chlorophylls a and b
A direct measurement of absorbance at 652, 665, and
750 nm in methanolic extracts was carried out using an Ultro-
spec 1100 pro-UV/visible spectrophotometer (GE Heathcare
Life Sciences, England). The following variables were calcu-
lated (in mg L−1 ): Chl a = (16.29 × OD665 ) − (8.54 × OD652 ) and Chl
b = (30.66 × OD652 ) − (13.58 × OD665 ). Each OD value was corrected
for the absorbance at 750 nm (e.g. OD665 = Abs665 − Abs750 ) (Porra
et al., 1989).
2.6. Total antioxidant activity
0.3 mL of methanolic extract sample has been mixed with 3.0 mL
reagent solution (0.6 M sulphuric acid, 28 mM sodium phosphate
and 4 mM ammonium molybdate). Reaction mixture has been incu-
bating at 95 ◦ C for 90 min under water bath. Absorbance of all
the sample mixtures was measured at 695 nm. Total antioxidant
activity is expressed as mg ascorbic acid × 100 g−1 of dry matter
(Ganesan et al., 2008).
2.7. Radical scavenging activity
The antioxidant activity of the polyphenolic extracts or standard
pure phenolic compounds was determined using 2,2-diphenyl-1-
picrylhydrazyl (DPPH) as a free radical. The reaction for scavenging
DPPH* radicals was performed in polypropylene tubes at room
temperature. Two milliliters of a 4 × 10−5 M methanolic solution
Family
Compositae
Compositae
Compositae
Compositae
Compositae
Compositae
of DPPH was added to 50 ␮L of the sample. The mixture was
shaken vigorously and left for 60 min. The absorbance of the result-
ing solution was measured at 517 nm. Methanol was used as a
blank solution, and DPPH* solution without any sample extract
served as control. The TEAC (Trolox equivalent antioxidant capac-
ity) values were calculated from the equation determined from
linear regression after plotting known solutions of Trolox with dif-
ferent concentrations (0.02–0.8 mM). The antiradical activity was
also expressed as the inhibition percentage and was calculated
using the following formula: % radical scavenging activity = (control
OD − (sample OD/control OD)) × 100 (Fattouch et al., 2007).
2.8. Determination of total phenolic compounds
Each methanol extract (0.2 mL) was mixed with 1.0 mL of
Folin–Ciocalteau’s reagent and 0.8 mL of saturated sodium carbon-
ate solution (7.5%). After 30-min incubation at room temperature,
the absorbance was read at 765 nm against a blank in an Ultro-
spec 1100 pro-UV/visible spectrophotometer (GE Healthcare Life
Sciences, England). Phenol content was calculated based on the cal-
ibration curves of gallic acid and expressed as g gallic acid × 100 g−1
of dry matter (Huang et al., 2006).
2.9. Determination of total flavonoid compounds
Each extract in methanol (0.5 mL) was mixed with 1.0 mL of 2%
methanolic AlCl3 ·6H2 O and the absorbance was measured 10 min
later at 430 nm. Content of flavonoids was calculated on the basis
of the calibration curves of quercetin, and was expressed as mg
quercetin × 100 g−1 of dry matter (Huang et al., 2006).
2.10. Quantification of flavonoids and phenolic acids profile
Methanol extracts were filtered through a 0.45-␮m filter. HPLC
analysis of flavonoids and phenolic acids was conducted in a
chromatograph (apparatus) Dionex liquid chromatography (USA)
equipped with a model P580 solvent pump (USA), a ASI-100
autosampler (USA), a PDA-100 photodiode array detector (USA) and
Dionex Software. A LiChrospher 100 RP-18 column (RP = reversed
phase) (25 cm × 4 mm, 5 ␮m; Merck, Darmstadt, Germany) was
used throughout this study. Phenolic acids and flavonoids were
detected at within 280 nm and 360 nm. The mobile phase was 5%
formic acid and methanol in a linear gradient starting at 15% and
reaching 35% in 15 min (then isocratic until 20 min), at a flow
rate of 1 mL/min and an injection volume of 20 ␮L. Phenolic com-
pounds were identified by comparison of their retention times with
those of pure standards and quantified individually based on stan-
dard curves of each flavonoid or phenolic acid type. Quantitation
was performed with the linear calibration curves of standard com-
pounds according to Jaakola et al. (2002, 2004).
2.11. Expression of data and statistical analysis
Results are the mean of three determinations. The results were
expressed as means ± standard deviation (SD). All statistical anal-
ysis was performed using the SPSS software package, version 16.0
(SPSS Inc., Chicago).
 I.S. Carvalho et al. / Industrial Crops and Products 33 (2011) 382–388 385

Table 2
Colour parameters (Tint, Colour Intensity, % Ye, % Rd and % Bl) and concentrations of chlorophylls a and b (mg g−1 of dry matter) from different Artimisia ssp. Leafs. Values
are means ± SD of triplicate assays.

Artimisia

annua arborescens ludoviciana oleandica princeps stelleriana

Chl a 42.69 ± 0.941a 2.61 ± 0.000b 24.45 ± 0.556c 28.65 ± 0.001c 23.43 ± 0.034c 34.33 ± 0.011c
Chl b 33.25 ± 0.564a 9.79 ± 0.123b 18.48 ± 0.450c 9.30 ± 0.002b 7.20 ± 0.002b 37.33 ± 0.001a
Colour intensity 4.47 ± 0.004a 4.12 ± 0.346a 5.00 ± 0.004a 4.63 ± 0.445a 8.08 ± 1.234b 5.24 ± 0.002a
Tint 0.21 ± 0.004a 0.17 ± 0.004a 0.19 ± 0.004a 0.19 ± 0.004a 0.22 ± 0.004a 0.19 ± 0.004a
Ye % 53.63 ± 0.02a 57.54 ± 0.445a 48.02 ± 0.054a 51.90 ± 0.123a 29.83 ± 2.345b 45.86 ± 0.004c
Rd % 20.93 ± 0.026a 24.40 ± 0.234a 26.66 ± 0.078a 24.34 ± 0.345a 36.34 ± 1.043b 27.75 ± 0.033a
Bl % 25.43 ± 0.045a 18.06 ± 0.004b 25.32 ± 0.000a 23.76 ± 0.230a 33.83 ± 0.894c 26.39 ± 0.056a

Means (±SD). Different letters in the same rows are significantly different at p < 0.05.

3. Results and discussion
3.1. Colour parameters, chlorophyll and total phenolic content
While growing six different Artemisia species, striking differ-
ences were observed between the leaves (Fig. 1). Leaves of plants
growing under the same greenhouse conditions were slightly
larger, for A. princeps spp. and A. stelleriana spp., with both the
blade and the petiole displaying a reddish colour. In the leaves
of A. annua spp., A. ludoviciana spp. and A. princeps spp., the red-
dish colour is not evident. This prompted us to examine colour
parameters, as well as the total phenolic and chlorophyll con-
tent of Artemisia leaves obtained from six different species. Colour
parameters were analyzed by direct measurement of absorbance
at 420, 520 and 620 nm in methanolic extracts. Results presented
in Table 2 indicate higher colour intensity (CI) in leaves of A. prin-
ceps spp. when compared with the other five leaves. In particular,
enhanced red colour in leaves of A. ludoviciana spp. and A. stelleri-
ana spp. (26.66% and 27.75% respectively) when compared with the
other leaves (20.93–24.40%) was observed. On the contrary, yellow
colour was reduced in those leaves (29.83% and 25.85%, respec-
tively, for A. ludoviciana spp. and A. stelleriana spp.). No differences
in blue colour were observed between leaves obtained from the
six Artemisia species, indicating a preponderant role for red and
yellow colours in those plants. Chlorophylls a and b content was
determined by direct measurements of absorbance at 652, 665 and
750 nm in methanolic extracts. Experimental results obtained show
that higher values for both chlorophylls (75.94 mg × 100 g−1 dry
matter) were obtained in leaves of A. annua spp.
Table 3 shows the phenolic compounds of six Artemisia leaves
expressed by mean (mg GAE × 100 g−1 of dry matter), which cor-
respond to the three analytical replicates. When separated by
dominant leaf colour, red pigmented Artemisia (A. stelleriana spp.
and A. ludoviciana spp.) show less (p < 0.05) phenolic (TPC) or
flavonoid (TFC) content when compared to green Artemisia.
3.2. Antioxidant capacity of Artemisia species
Consistent with the assay of TPC, DPPH scavenging activity of red
Artemisia (quenched 62.44% and 75.78% radicals in the system) was
not significantly higher (p < 0.05) than green Artemisia (quenched
between 77.82% and 80.60% radicals in the system). An exception
was observed for A. annua spp., quenched 36.82% radicals in the
system (reflecting a 2-fold difference). The overall mean value of
the DPPH scavenging activity of the Artemisia species was 70% DPPH
quenched. With the highest value for the green A. arborescens spp.
and the lowest for the green A. annua spp. reflecting about 50%
difference over the 2 species (Table 3).
When compared in a linear correlation model, the TPC was
highly correlated with DPPH scavenging ability of Artemisia
(r2 = 0.968) (Fig. 2a). This method is recommended by many authors
(Klimczak et al., 2007; Shahidi et al., 2006) as easy and accu-
rate assays for measuring the antioxidant activity of food samples.
The DPPH scavenging activity of the Artemisia leaves extracts was
determined and found in proportion to the Trolox concentration,
suggesting that all Artemisia leaves (with exception to A. annua
spp. and A. stelleriana spp.) have a very good antioxidant capac-
ity (Fig. 2b). In fact, the correlation between antioxidant capacity
and the total phenolic content in others plants has already been
reported by Fattouch et al. (2007). In the present study the Artemisia
leaves exhibited a stronger antioxidant effect when compared
to the Trolox concentration (Fig. 2b). Probably the differences
between the overall antioxidant capacities could result from dif-
ferent interactions between different antioxidant components.
3.3. Flavonoid and phenolic acid profile
The profile of polyphenolic compounds in the leaves of Artemisia
species analyzed is shown in Table 4. The range of total phenol con-
tent was 0.22–0.39 mg of GAE g−1 of dry matter for the Artemisia
spp. leaves. A total of 18 polyphenolic compounds were identi-
fied and quantified using high-performance liquid chromatography
combined with a photodiode-array detector (HPLC-PDA), showing
the separation of hydroxybenzoic acids (4), hydroxycinnamic acids
(5), flavonols (3), and catechins (2) compounds (Fig. 3).
Flavonols, catechins and kaempferol compounds present in veg-
etables have been considered a therapeutic agent due to their
beneficial health effects, such as their supposed protection against
certain cancers, cardiovascular diseases and aging (Ross and Kasum,
2002). Moreover, they have also been deemed a source of colours

Table 3
Total phenolic compound (g GAE × 100 g−1 of dry matter), total flavonoids (mg quercetin × 100 g−1 of dry matter), total antioxidant activity (mg ascorbic acid equivalents.
100 g−1 of dry matter) and radical scavenging activity (%) from different Artimisia ssp. Leafs. Values are means ± SD of triplicate assays.

Artimisia

annua arborescens ludoviciana oleandica princeps stelleriana

Total phenolic compounds 0.22 ± 0.002a 0.23 ± 0.005a 0.36 ± 0.000b 0.39 ± 0.001b 0.39 ± 0.000b 0.39 ± 0.004b
Total flavonoids compounds 0.14 ± 0.003a 0.03 ± 0.005b 0.10 ± 0.010a 0.11 ± 0.004a 0.19 ± 0.002a 0.19 ± 0.002a
Total antioxidant activity 0.36 ± 0.074a 0.24 ± 0.034b 0.42 ± 0.014c 0.49 ± 0.023c 0.21 ± 0.022b 0.53 ± 0.084c
Radical scavenging activity 36.82 ± 0.145a 80.60 ± 0.366b 75.78 ± 0.367b 77.82 ± 0.023b 79.45 ± 0.026b 62.44 ± 0.634c

Means (±SD). Different letters in the same rows are significantly different at p < 0.05.
386 I.S. Carvalho et al. / Industrial Crops and Products 33 (2011) 382–388

Fig. 2. (a) Linear correlation between the radical scavenging activity (%) of Trolox and total antioxidant activity (mg ascorbic acid equivalents × 100 g−1 of dried matter)
for methanolic extracts. Correlation coefficient (r) = 0,984 and coefficient of determination. (r2 ) = 0.968. (b) Radical scavenging activity (%) of Trolox used at equivalent
concentrations as in the different Artimisia ssp. leafs extracts.

for food products, mainly anthocyanins, in alternative to synthetic
dyes whose harmful effects upon human health have often been
assumed and, in some cases, demonstrated. A. annua spp. leaves
had 80 ␮g g−1 of dry matter of catechins, 2 ␮g g−1 of dry matter of
flavonols, 75 mg g−1 of dry matter of hydroxycinnamic acids and
430 ␮g g−1 of dry matter of hydroxybenzoic acids. The others five
species, having the same number of total phenolic compounds,
contained between 19 ␮g g−1 of dry matter and 936 ␮g g−1 of
dry matter of catechins, between 2 ␮g g−1 of dry matter of and
54 ␮g g−1 of dry matter of flavonols, between 605 ␮g g−1 of dry
matter and 1575 ␮g g−1 of dry matter of hydroxycinnamic acids
and between 41 ␮g g−1 of dry matter and 512 ␮g g−1 of dry mat-
ter of hydroxybenzoic acids. Two hydroxybenzoic acids, gallic acid
and benzoic acid were detected in five Artemisia leaves. The major
hydroxybenzoic acid content in all Artemisia leaves (obtained for A.
ludoviciana spp.) is gallic acid (3,4,5-tri hydroxybenzoic acid). Gallic
acid is a naturally abundant plant phenolic compound. It is present
in food of plant origin, and since it was found to exhibit antiox-
idative properties, it has attracted considerable interest. The five
hydroxycinnamic acids identified in the analysis were p-coumaric
acids, caffeic and ferulic acid conjugates, chlorogenic acid, sinapic
acid and cinnamic acid. Caffeic acid and ferulic acid conjugates were
the most dominant hydroxycinnamic acids in all Artemisia leaves
studied (Table 4) as it accounted for the largest proportion of the
total hydroxycinnamic acids contents. Sinapic acid was the second
most abundant hydroxycinnamic acid and followed by chlorogenic,
coumaric, caffeic acids in all Artemisia species. The levels of caffeic
and ferulic acid conjugates and sinapic acids in the Artemisia species
were slightly lower than those reported by Jaakola et al. (2004) in
sweet potato leaves.
The qualitative and quantitative compositions of flavonols also
differed within leaves of plants growing under the same conditions.
Kaempferol, the major flavonol in the Artemisia leaves studied, was
detected in higher amounts, 47.56 ␮g g−1 of dry matter, in A. ole-
andica, whereas quercetin and myricetin were only detected in
much lower amounts, probably because high levels of kaempferol,
produced upstream in the metabolic pathway, prevent accumula-
tion of these compounds. Previous work has established that the
antioxidant properties of some plants are partly due to low molec-
ular mass phenolic compounds, particularly flavonoids, which are
known to be potent antioxidants (Wang et al., 1999). The results
suggest that flavonols like kaempferol and the catechins, together
with hydroxycinnamic (caffeic and ferulic acid conjugates) and
hydroxybenzoic acids (gallic acid), play a predominant role in the
growing leaves. In humans, the presence of flavonoids may con-
tribute to the neutralization of cell-damaging free radicals and the

Table 4
Concentrations ((g g−1 of dry matter) of flavonols and hydroxycinnamic acids in different leaves of Artimisia spp.. Values are means ± SD of triplicate assays.

Compounds Artimisia

annua arborescens ludoviciana oleandica princeps stelleriana

Catechins
(+)-Catechin 79.53 ± 0.123a 19.75 ± 0.003b 1.67 ± 0.123c 935.66 ± 23.009d 79.40 ± 2.023a ND
(−)-Epicatechin 0.74 ± 0.034a ND 21.28 ± 1.345b ND 5.31 ± 0.345c ND
Total 80.27 19.75 22.95 935.66 84.71 ND
Flavonols
Quercetin 0.74 ± 0.004a ND 7.63 ± 0.123b 4.68 ± 0.145c ND 1.25 ± 0.133 d

Myricetin 0.74 ± 0.089a 4.43 ± 0.344b 1.13 ± 0.123a 1.23 ± 0.081a ND 0.86 ± 0.001a
Kaempferol 0.74 ± 0.005a 4.68 ± 0.045b 1.80 ± 0.123c 47.56 ± 0.345d 31.60 ± 1.440e ND
Total 2.22 9.11 10.56 53.47 31.60 2.11
Hydroxycinnamic acids
p-Coumaric conjugates 17.23 ± 1.022a ND ND 24.27 ± 0.072b ND ND
Caffeic or ferulic conjugates 52.49 ± 1.046a 1275.71 ± 0.788b 798.65 ± 13.056c 730.42 ± 0.034c 1308.96 ± 22.344b 562.95 ± 12.334d
Chlorogenic 0.76 ± 0.022a ND 16.80 ± 1.671b ND ND 41.19 ± 2.001c
Sinapic 3.26 ± 0.722a 284.67 ± 0.124b 5.95 ± 0.451a 40.61 ± 0.034c 17.36 ± 1.333d 0.87 ± 0.033e
Cinnamic 0.74 ± 0.043a 14.73 ± 0.456b 6.06 ± 0.143c 3.44 ± 0.046c 1.28 ± 0.222a ND
Total 74.48 1575.11 827.46 798.74 1327.60 605.01
Hydroxybenzoic acids
Gallic 70.96 ± 1.054a 266.46 ± 0.136b 496.32 ± 8.777c 265.39 ± 0.097b 334.23 ± 10.555d 38.45 ± 0.233e
Benzoic 0.74 ± 0.043a ND 7.41 ± 1.665b 22.27 ± 0.041c 38.44 ± 2.934d 1.09 ± 0.002a
Gentisic 357.92 ± 11.054a ND 9.20 ± 0.888b ND ND 1.28 ± 0.001c
p-Anisic 0.74 ± 0.022a ND 1.37 ± 0.056a 1.26 ± 0.001a ND ND
Total 430.36 266.46 512.93 288.92 372.67 40.82

Means (±SD). Different letters in the same rows are significantly different at p < 0.05. ND, not detected.
 I.S. Carvalho et al. / Industrial Crops and Products 33 (2011) 382–388
Fig. 3. HPLC chromatograms examples at 278, 310, 326 and 348 of different
flavonoids and phenolic acids from leaves of A. ludoviciana ssp. Peaks: 1 – gallic
acid, 2 – gentisic, 3 – catechin, 4 – chlorogenic acid, 5 – caffeic acid, 6 – epicatechin,
7 – p-coumaric acid, 8 – sinapic acid, 9 – benzoic acid, 10 – anisic acid, 11 – kamferol,
12 – myricetin, 13 – cinnamic acid and 14 – quercetin.
387
maintenance of heart health (Ross and Kasum, 2002). The pres-
ence of hydroxycinnamic and hydroxybenzoic acids in our diets
may also contribute to bolster cellular antioxidant defenses and to
the maintenance a healthy vision. Although flavonoids are increas-
ingly recognized as playing important roles as antioxidants, further
work is necessary to uncover the full potential of these compounds
in the improvement of human health, as well as to identify their
presence in the various foods and elucidate environmental effects
on their content in plants.
4. Conclusions
According to the data obtained in the present study, the total
phenolic and flavonoid contents of the six Artemisia species did sig-
nificantly differ with A. annua and A. stelleriana showing about 50%
less amount when compared with the other four species. Ferulic
and caffeic conjugates acid was the most dominant hydroxycin-
namic acid and catechin was the most dominant flavonoid in all
six Artemisia species. The antioxidant capacity of A. annua and A.
stelleriana leaves was found lower, reflecting a 2-fold difference
lower than the other four species used in this study highly related
with total phenolic compounds. The knowledge of the phenolic pro-
file, occurring in Artemisia spp., holds great significance from both
dietary and nutritional point of view.
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