ARTICLE IN PRESS

JOURNAL OF
FOOD COMPOSITION
AND ANALYSIS
Journal of Food Composition and Analysis 21 (2008) 107–115
www.elsevier.com/locate/jfca

Original Article

Anthocyanin composition in fig (Ficus carica L.)

Montserrat Dueñas, José Joaquı́n Pérez-Alonso, Celestino Santos-Buelga,
Teresa Escribano-Bailón
Grupo de Investigación en Polifenoles, Unidad de Nutrición y Bromatologı´a, Facultad de Farmacia, Universidad de Salamanca,
Campus Miguel de Unamuno, 37007 Salamanca, Spain
Received 16 March 2007; received in revised form 5 September 2007; accepted 13 September 2007

Abstract

The anthocyanin composition was analysed in fig fruit (Ficus carica L.) from five different varieties (Colar, Cuello de Dama (green),
Cuello de Dama (dark purple), Granilla and Bursa Siyahi). Fifteen anthocyanin pigments were detected, most of them containing
cyanidin (Cy) as aglycone; some pelargonidin (Pg) derivatives were also found. Rutinose and glucose were present as substituting
sugars, as well as acylation with malonic acid. Minor levels of peonidin 3-rutinose (Pn 3-rutinoside) in the pulp were also detected.
Other noticeable aspects in the pigment composition of the fig were the detection of anthocyanidin-derived pigments, namely
5-carboxypyranocyanidin-3-rutinoside, a cyanidin 3-rutinose dimer and five condensed pigments containing C–C linked anthocyanins
(Cy and Pg) and flavanol (catechin and epicatechin) residues. Total anthocyanin content in the skin ranged between 32 and 97 mg g 1 and
between 1.5 and 15 mg g 1 in the pulp. The main anthocyanin in both parts of the fruit was Cy 3-rutinoside (48–81% in skin and 68–79%
in pulp) usually followed by Cy 3-glucoside (5–18% in skin and 10–15% in pulp). Malonyl derivatives were more abundant in the skin
(1.2–6.5%) than in the pulp (1.0–2.6%).
r 2007 Elsevier Inc. All rights reserved.

Keywords: Anthocyanins; Ficus carica L.; Fig fruit; Cyanidin; Anthocyanin-derived pigments; Fig skin; Fig pulp; Fig varieties; HPLC

1. Introduction and dried figs also present relatively high amounts of crude
fiber (5.5%, w/w) and polyphenols (Vinson, 1999; Vinson
The Common Fig (Ficus carica L.) is a tree native to et al., 2005).
southwest Asia and the eastern Mediterranean region, Some recent works have reported that fig antioxidants
belong to botanical family Moracea. The Common Fig is can protect lipoproteins in plasma from oxidation and
one of the first plants that was cultivated by humans and is produce a significant increase in plasma antioxidant
an important crop worldwide for dry and fresh consump- capacity for 4 h after consumption (Vinson et al., 2005).
tion. Its edible part is commonly referred to as a ‘‘fruit’’ Also, Solomon et al. (2006) showed that the higher the
although it is a synconium, that is, a fleshy, hollow polyphenols contents, especially anthocyanins, in fig fruit,
receptacle with a small opening at the apex partly closed by the higher was their antioxidant activity.
small scales. According to our knowledge, there are no studies about
Previous reports concerning the nutrient composition of the detailed pigment composition in fig. Early studies
dried figs have indicated that it has the best nutrient score carried out by Robinson and Robinson (1932) identified
among the dried fruit, being an important source of cyanidin 3-glucoside (Cy 3-gluc). Furthermore, four
minerals and vitamins (USDA, 2002). The presence of anthocyanin were reported in the fig, with cyanidin
phytosterols (433 mg/100 g dry basis) has also been 3-rhamnoglucoside (Cy 3-rut) accounting for about
reported in fig fruit (Jeong and Lachance, 2001). The fresh 75% of total pigments; other pigments were cyanidin
3,5-diglucoside (11%), cyanidin 3-glucoside (11%) and
Corresponding author. Tel.: +34 923 29 45 37; fax: +34 923 29 45 15. pelargonidin 3-rhamnoglucoside (3%) (Puech et al., 1975;
E-mail address: mduenas@usal.es (M. Dueñas). Solomon et al., 2006; del Caro and Piga, 2007).

0889-1575/$ - see front matter r 2007 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2007.09.002
 ARTICLE IN PRESS
108 M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115

The contents of total anthocyanins, polyphenols and three independent extracts were prepared, purified and
flavonoids in the skin and pulp from commercial varieties analysed separately.
of figs with different colour (black, red, yellow and green)
have been recently analysed by Solomon et al. (2006). The 2.3. HPLC–diode array detector (DAD)–MS analyses
fruit skin contributed most of the polyphenols and
antioxidant activity compared to the pulp especially in The anthocyanin extracts were analysed using a Hewlett-
darker varieties. Packard 1100 series liquid chromatograph (Agilent Tech-
The aim of the present work was to characterise the nologies, Waldbronn, Germany). Separation was achieved
qualitative and quantitative pigment composition, in the on an AQUAs (Phenomenex, Torrance, CA) reverse-
skin and flesh of five different fig varieties, using HPLC phase C18 column (5 mm, 150 mm  4.6 mm i.d.) thermo-
coupled to diode array and mass spectrometer (MS) statted at 35 1C. The solvents used were: (A) 0.1% TFA in
detection. water and (B) 100% HPLC-grade acetonitrile. The
gradient employed was: isocratic 10% B for 3 min,
from 10% to 15% B for 12 min, isocratic 15% B for
2. Materials and methods
5 min, from 15% to 18% B for 5 min, from 18% to 30% B
for 20 min and from 30% to 35% for 5 min, at a flow
2.1. Samples
rate of 0.5 mL min 1. Detection was carried out in a
DAD, using 520 nm as the preferred wavelength, and
Fig fruit (F. carica L.) from five selected commercial
in a MS connected to the HPLC system via the DAD
varieties: cv. Colar, Cuello de Dama (green), Cuello de
cell outlet.
Dama (dark purple), Granilla and Bursa Siyahi, were
LC–MS analyses were performed using a FinninganTM
sampled from five retail outlets in Salamanca (Spain). They
LCQ MS detector (Thermoquest, San Jose, CA) equipped
were harvested in 2006 at the commercial mature stage in
with an API source, using an electrospray ionisation (ESI)
different Spanish regions excepting the Bursa variety from
interface. Both the sheath gas and the auxiliary gas were
Turkey. Approximately 1 kg of each variety was randomly
nitrogen at flow rates of 1.2 and 6 L min 1, respectively.
selected from bins in each retail outlet; 2–5 units were
The capillary and source voltage were 4 V and 4.5 kV,
randomly taken for each extraction (three extractions for
respectively, and the capillary temperature was 195 1C.
each variety). The figs were weighed and immediately
Spectra were recorded in positive ion mode between m/z
peeled. Skin and pulp were weighed separately and
120 and 1500. The MS was programmed to carry out a
extracted.
series of three consecutive scans: a full mass, an MS2 scan
of the most abundant ion in the full mass, and an MS3 of
2.2. Sample preparation the most abundant ion in the MS2, using a normalised
energy of collision of 45%.
The fruit (2–5 units) were manually peeled and the skin
separated from the pulp and analysed separately. The yield 2.4. Quantification
of the skin versus the entire fruit was 5–6%. The skin
(3–5 g) and pulp (20–30 g) were ground and homogenised For the quantitative analysis of anthocyanins, a calibra-
in MeOH containing 0.5% trifluoracetic acid (TFA), using tion curve was obtained by injection of different concen-
a Polytron homogeniser (Kinematica, Littau, Switzerland) tration of cyanidin 3-O-glucoside (for cyanidin-based
and macerated for 16 h at 4 1C. Subsequently, they were anthocyanins), pelargonidin 3-O-glucoside (for pelargoni-
centrifuged for 20 min at 4000  g at 5 1C in a refrigerated din-based anthocyanins) and peonidin 3-O-glucoside (for
superspeed centrifuge (Sorvall RC 5B). This process was peonidin-based anthocyanins) standards purchased from
repeated twice at two intervals of 4 h, completing a total Polyphenols Labs., Sandnes, Norway.
of 24 h of extraction. The extracts were combined and The limit of detection (LOD) and limit of quantification
concentrated under vacuum at 30 1C until the methanol (LOQ), respectively, defined as a 3:1 and 10:1 peak to noise
was removed. The final volume was adjusted to 50 mL. The ratio, were determined for cyanidin 3-O-glucoside, pelar-
aqueous extract was stored at 30 1C until purification and gonidin 3-O-glucoside and peonidin 3-O-glucoside. The
analysis. For purification, an aliquot (2 mL) of the aqueous working concentration range was 0.001 and 0.1 mg mL 1
extract was deposited onto a C18 Sep-Pak cartridge for three compounds. The calibration curves were calcu-
(Waters, Milford, MA, USA), previously activated with lated by plotting peak area ratio (Y) versus concentration
methanol followed by water; sugars and more polar (X, mg mL 1) of the three anthocyanins in the standard
substances were removed by passing 10 mL of ultrapure solution at six different concentrations, with least-squares
water and anthocyanins pigments were eluted with 4 mL of linear regression. Within this interval, the calibration
0.5% TFA in MeOH. Afterwards, the methanolic extract curves were linear with correlation coefficient for cyanidin
was evaporated under vacuum in a rotary evaporator 3-O-glucoside, pelargonidin 3-O-glucoside and peonidin
(30 1C) and then dissolved in aqueous 0.1% TFA for 3-O-glucoside (R240.999). The limits of detection were
HPLC analysis. For the skin and pulp of each variety, calculated under the Glaser criteria (Glaser et al., 1981).
 ARTICLE IN PRESS
M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115 109

The limits of detection were 0.016 mg mL 1 for Cy 3-O- 3. Results and discussion
glucoside, 0.025 mg mL 1 for Pg 3-O-glucoside and
0.014 mg mL 1 for Pn 3-O-glucoside. Limits of quantifica- 3.1. Pigment identification
tion were 0.048 mg mL 1 for Cy 3-O-glucoside,
0.074 mg mL 1 for Pg 3-O-glucoside and 0.042 mg mL 1 Up to 15 anthocyanin pigments were detected in the fig
for Pn 3-O-glucoside. The precision of the chromato- samples analysed. As an example, Fig. 1 shows the HPLC
graphic method was satisfactory. Six replicate determina- pigment profile in extracts from skin and pulp of Colar fig
tions of the standards were carried out on the same day. variety. Data (retention time, lmax in the visible region,
Relative standard deviations were calculated with results of molecular ion and main fragment ions observed in MS2
coefficients of variation less than 7%. and MS3) obtained for the anthocyanin peaks in the
HPLC–DAD–MS analyses are presented in Table 1,
2.5. Statistical analysis together with their occurrence in pulp and skin. Pigments
were cyanidin (Cy) or pelargonidin (Pg) derivatives, as
The determination of anthocyanin contents in skin and demonstrated from their UV–vis spectra and mass spectral
pulp extracts of each variety were carried out in triplicate data, only compound 14 showed data in concordance with
and the results are given as mean7standard deviation peonidin as aglycone (lmax at 520 nm and MS3 at m/z 301).
(S.D.). Data were analysed by one-way analysis of variance Peaks 4, 10, 11 and 13 were usually the major pigments
(ANOVA). Significant differences were assessed with an identified in the skin, which were identified as Cy 3,5-
LSD test (po0.05). The statistical analysis was performed diglucoside (Peak 4), Cy 3-glucoside (Peak 10), Cy
using PC software package SPSS (version 13.0; SPSS Inc., 3-rutinoside (Peak 11) and Pg 3-rutinoside (Peak 13). The
Chicago). identity of these compounds was elucidated by comparison

10 11
140

120
13

100
4
mAU

80

60 3
8
40 1
15

20 7 9 12
2 5
6

0
0 10 20 30 40 50
min

8 10 11
140
4
120
3
100
mAU

80

60

40 13
5
2 14 15
20 9 12

0
0 10 20 30 40 50
min

Fig. 1. Chromatograms (zoom) recorded at 520 nm showing the anthocyanin profiles of fig fruit (variety: Colar). (A) Skin fig, (B) pulp fig.
 ARTICLE IN PRESS
110 M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115

Table 1
Retention time (Rt), wavelengths of maximum absorption in the visible region (lmax), mass spectral data and tentative identification of anthocyanins
pigments in fig skin and pulp

Peak Rt lmax Molecular ion MS2 (m/z) MS3 (m/z) Tentative identification
(min) (nm) [M+H]+ (m/z)

1 12.9 528 1189 881 735, 573, 421, 287 Cy 3-rutinoside dimer
2 13.6 528 737 575, 557 557, 449, 423, 329, 287 (Epi)catechin-(4-8)-Cy 3-glucoside
3 14.4 528 883 737, 575 557, 449, 423, 329, 287 (Epi)catechin-(4-8)-Cy 3-rutinoside
4 15.2 516 611 449, 287 287 Cy 3, 5-diglucoside
5 16.8 528 883 737, 575 557, 423, 329, 287 (Epi)catechin-(4-8)-Cy 3-rutinoside
6 17.5 524 867 721, 559 559, 541, 433, 407, 312 (Epi)catechin-(4-8)-Pg 3-rutinoside
7 19.1 525 867 721, 559 559, 541, 433, 407, 312 (Epi)catechin-(4-8)-Pg 3-rutinoside
8 20.4 506 663 355 327, 311, 299 5-Carboxypyranocyanidin-3-rutinoside
9 20.8 520 697 535, 449, 287 287 Cy 3-malonylglicosyl-5-glucoside
10 22.2 516 449 287 287 Cy 3-glucoside
11 22.8 518 595 449, 287 287 Cy 3-rutinoside
12 27.7 n.a. 433 271 – Pg 3-glucoside
13 28.6 506 579 433, 271 271 Pg 3-rutinoside
14 29.9 520 609 463, 301 301 Pn 3-rutinoside
15 32.9 518 535 287 287 Cy 3-malonylglucoside

n.a.: not available.

of their chromatographic characteristics and absorption malonyl-hexoside residue) and m/z 287 (cyanidin), was
spectra with data in our library and confirmed by mass consistent with Cy 3-malonylglucosyl-5-glucoside. Further
analysis (Table 1). Compounds 4, 10 and 11 showed lmax at confirmation of the identity of compounds 9 and 15 was
516–518 nm characteristic of cyanidin derivatives and provided by comparison of their characteristics with those
compound 13 at 506 nm together with a spectrum shape of the same compounds previously identified in our
characteristic of pelargonidin derivatives (Santos-Buelga laboratory in purple corn and strawberry (de Pascual-
et al., 2003). The presence of cyanidin and pelargonidin as Teresa et al., 2002; Lopes da Silva et al., 2007) and
anthocyanidins in these peaks was confirmed by their mass available in our anthocyanin library.
spectra, which showed MS2 signals at m/z [M]+ 287 and Compounds 2, 3, 5, 6 and 7 were assigned to condensed
271, respectively. The MSn fragmentation profiles were in pigments containing C–C linked anthocyanin (Cy or Pg) and
accordance with the proposed identities of the compounds. catechin residues. Pigments with similar structural character-
These four anthocyanins had been described in fig by other istics were found in other sources as beans (Macz-Pop et al.,
authors (Robinson and Robinson, 1932; Puech et al., 1975; 2006; González-Paramás et al., 2006), purple corn extracts
Solomon et al., 2006, del Caro and Piga, 2007). However, (de Pascual-Teresa et al., 2002), strawberries (Fossen et al.,
to the best of our knowledge, the rest of identified pigments 2004; Lopes da Silva et al., 2007), red wine (Salas et al., 2004)
that will be discussed below are here described in fig for the and black currant (McDougall et al., 2005). These com-
first time. pounds presented bathochromic shifts of about 12 nm in the
Compound 12 was identified as Pg 3-glucoside by visible region of their absorption spectra with regard to the
comparison of its chromatographic, spectral and MS parent anthocyanins (see Table 1), as also found by other
characteristics with those of a commercial standard. Peak authors (Fossen et al., 2004; Salas et al., 2004; Lopes da Silva
14 was only detected in the pulp and showed a molecular et al., 2007).
ion at m/z 609. Its MS2 gave two fragments at m/z 463 Peak 2 showed a positive molecular ion [M]+ at m/z 737,
([M 146]+, loss of a rhamnose moiety) and at m/z 301 with a major MS2 fragment at m/z 575 ([M 162]+, loss of
([M 308]+ loss of rhamnoglucoside moiety). This an hexose moiety). Its MS3 fragmentation led several
fragmentation pattern is characteristic of rutinosides fragments ions at m/z 557 ([M 18]+, loss of water), m/z
(Giusti et al., 1999) and the compound was identified as 449 ([M 126]+, loss of a C6H6O3 residue from A ring), m/z
peonidin 3-O-rutinoside. 423 ([M 152]+ retro Diels-Alder cleavage (RDA) of a
Peaks 9 and 15 corresponded to acyl derivatives of flavanol unit), m/z 329 ([M 246]+ partial loss of (epi)ca-
cyanidin. Peak 15 showed lmax at 518 nm and spectra techin, m/z at 287 ([M 288]+, loss of an upper (epi)ca-
molecular ion at m/z 535 releasing a unique MS2 fragment techin unit. This type of fragmentation profile was
at m/z 287 ([M 248]+, loss of a malonylglycoside moiety). consistent with the one previously described for anthocya-
These characteristics allowed its identification as Cy 3-O- nin–catechin condensed pigments (González-Paramás
malonylglucoside. For peak 9, the molecular ion at m/z et al., 2006).
697, releasing three fragments MS2 at m/z 535 ( 162 amu, Peaks 3 and 5 showed identical molecular ions (m/z [M]+
loss of an hexose), m/z 449 ( 162–86 amu, loss of a at 883) and fragmentation patterns. Fig. 2 shows the MS2
 ARTICLE IN PRESS
M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115 111

883.2 575.1 - 308 (rhamnoglucoside)

100 100
MS2
90 90
80 80
Relative Abundance

Relative Abundance
70 70
60 60
50 50
40 884.2 40
30 30 - 146 (rhamnose)
20 20 423.1
557.2 737.1
10 329.1 883
345.0 885.2 10
287.2 449.1
0 0
200 400 600 800 1000 1200 1400 300 400 500 600 700 800
m/z m/z

100 -152 (RDA)

423.0
MS3
90
80

Relative Abundance
70 -288 (catechin)
60
329.1
50 -246

40
-18 (H2O)
557.1
30
20
287.3 575
361.4
10
0
200 300 400 500
m/z
OH
OH OH
OH
O OH
OH OH

OH
OH O
OH O
OH OH
OH -308 (rhamnoglucoside) OH
OH
O+
OH OH
OH
-18 (H2O) OH
OH OH
HO OH
O O+
O OH OH O+
OH OH
HO
O
-162 amu OH OH
OH
O OH
OH
-152 (RDA)
OH
-146 amu CH3

-246
-288 OH O
OH OH
OH
OH OH OH
OH

O+ OH
O+
OH OH O+
OH

OH OH
OH
OH OH
OH

Fig. 2. MS2 and MS3 of peak 3: (epi)catechin-(4-8)-Cy 3-rutinoside (m/z [M+] at 883).

and MS3 spectra of pigment 3 together with its fragmenta- and rhamnoglucoside moieties. MS3 fragmentation of the
tion scheme. The MS2 fragments at m/z 737 ([M 146]+) ion at m/z 575 gave the same fragments as detected for
and 575 ([M 308]+) corresponded to the loss of rhamnose pigment 2 (Table 1). Thus, these compounds were
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112 M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115

identified as pigments resulting from the direct condensa- 3.2. Content and distribution of anthocyanin pigments in fig
tion of Cy 3-O-rutinoside and either catechin or epicate- varieties
chin.
Peaks 6 and 7 showed lmax in the visible region at The content and distribution of anthocyanins were
524 nm, bathochromically shifted with regard to that of the investigated in five fig varieties. One of them a green
precursor anthocyanin (Pg 3-O-glucoside, lmax at 506 nm). variety, in which the content was only investigated in its
These compounds showed a positive molecular ion [M]+ at brown–pink pulp. The other varieties presented an intense
m/z 867, with an MSn fragmentation characteristic of violet–blue colour in the skin and dark pink colour in the
anthocyanin–catechin condensed pigments. MS2 were flesh. In these, both pulp and skin were analysed.
found at m/z 721, ([M 146]+, loss of a rhamnose moiety), Significant differences in the anthocyanin content were
and 559 ([M 308]+, loss of a rhamnoglucoside moiety). found between pulp and skin and also among some
MS3 fragmentation of the ion at m/z 559 gave signals at m/z varieties. Table 2 shows the individual concentration of
541 ([M 18]+, loss of water), m/z 433, ([M 126]+, loss of the anthocyanin pigments (mg g 1 fresh weigh) found in the
a C6H6O3 residue), m/z 407 ([M 152]+, RDA of the skin of the fig varieties and Table 3 those of the pulp. The
flavanol unit), m/z 312 ([M 246]+, partial loss of concentration of anthocyanins in the skin ranged from 32
(epi)catechin, m/z 271 ([M 288]+, loss of (epi)catechin and 97 mg g 1 fresh weight. Granilla, Cuello de Dama (dark
unit. Thus, compounds 6 and 7 were identified as purple) and Bursa Siyahi were the varieties with the higher
(epi)catechin-Pg 3-O-rutinoside condensed pigments. Com- anthocyanin content without significant differences be-
pounds consisting of (epi)afzelechin-Pg 3-O-rutinoside tween them. Colar variety had the minor content. In the
have been described in strawberry (Lopes da Silva et al., pulp, the anthocyanin contents ranged from approximately
2007), but according to our knowledge this is the first time 14 mg g 1 for Colar and Cuello de Dama (dark purple)
that similar compounds containing (epi)catechin as flava- varieties to 1.5 mg g 1 for Cuello de Dama (green), this later
nol moiety are reported. expected according with its fair red colour. Solomon et al.
Peak 1, showed a positive molecular ion [M]+ at m/z (2006) found contents of the same order for the variety
1189 and its MS2 fragmentation led one fragment at m/z Bursa, 41 mg g 1 for skin and 1 mg g 1 for pulp.
881 ([M 308]+) corresponding to the loss of a rhamno- Regarding anthocyanin distribution, both in skin and in
hexoside moiety. MS3 fragmentation of this ion gave pulp, the cyanidin derivatives were the most abundant. Cy
fragments at m/z 735 and 573 corresponding to successive 3-rutinoside was the predominant compound in (48–81%
losses of rhamnose ( 146 amu) and rhamnoglucoside in the skin and 68–79% in the pulp), followed by Cy 3-
( 308 amu) moieties from a Cy 3-rutinoside unit. Further glucoside (5–18% in the skin and 10–15% in the pulp)
information to conclude about the identity of compound 1 except in the skin of Collar variety where Cy 3,5-digluco-
was provided by other fragment detected at m/z 421, side and Pg 3-rutinoside were more abundant (11–12%).
( 152 amu) corresponding to the RDA fission of a Percentages of 75% for Cy 3-rutinoside, 11% for Cy 3,5-
cyanidin unit. This fragmentation pattern is similar to the diglucoside, 11% for Cy 3-glucoside and 3% for Pg 3-
one obtained by Vidal et al. (2004) for malvidin 3-glucoside rutinoside were found by Puech et al. (1975) in a study
dimer isolated from grape skins. Thus, the compound was carried out with the variety Mission (pulp and skin).
tentatively interpreted as a cyanidin 3-rutinoside dimer, for Malonyl derivatives were also found being usually more
which two structures could be proposed, an A-type flavan- abundant in skin (1.2–6.5%) than in pulp (1.0–2.6%). They
flavylium and a B-type flavene-flavylium (Fig. 3). were particularly abundant in the skin of the Granilla
Peak 8 showed a molecular ion at m/z 663. Its MS2 variety.
fragmentation gave one fragment at m/z 355 ([M 308]+, It was noticeable the presence of Pg derivatives in the
loss of a rhamnohexoside moiety). The MS3 fragmentation skin extracts of Colar variety which percentages around
of this ion gave, among others, a minor fragment at m/z 13% (Pg 3-glucoside+Pg 3-rutinoside). In other varieties,
311 ( 44 amu) corresponding to a loss of a carboxyl these compounds ranged between 0.8% and 2.2%.
residue, allowing its tentative identification as carboxypyr- Peonidin 3-rutinoside was observed in pulp extracts of all
anocyanidin 3-O-rutinoside. This compound showed lmax the studied varieties, however in the skin no evidences of
in the visible region at 506 nm, hypsochromically shifted this compound were found when the extracts were screened
with regard to that of the precursor anthocyanin (Cy 3-O- for its ion (m/z 609) in the full spectra of the LC–MS
glucoside, lmax at 516 nm). chromatograms.
The same compound was described in processed black The levels of Cy 3-rutinose dimer were in general scarce.
olives (del Caro et al., 2006). This type of pyranoantho- This compound was not detected in the pulp except in the
cyanin pigments have been mostly associate to reactions variety Cuello de Dama (dark purple). Colar skin showed
involving anthocyanins during processing of plant-derived the greatest content of this compound with almost 3% of
products, such as red wines (Fulcrand et al., 1998; Bakker the anthocyanins identified in this variety. Furthermore,
et al., 1997), although they have also been reported in non- this variety presented the greatest content in flavanol–
processed fruit like strawberry (Lopes da Silva et al., 2007; anthocyanin condensed pigments (2.6 mg g 1 that repre-
Andersen et al., 2004). sents 8.2% of total anthocyanins in the skin, and 0.8 mg g 1
 ARTICLE IN PRESS
M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115 113

1189.2 881.1 - 308 (rhamnoglucoside)

100 100
MS2
90 90
80 80
Relative Abundance

Relative Abundance
70 70
60 60
50 50
718.9
40 40
30 30
1191.3 1189
20 20
10 275.7 10
0 0
200 400 600 800 1000 1200 1400 400 500 600 700 800 900 1000 1100 1200
m/z m/z

735.1 - 146 (rhamnose) OH

100 OH
MS3 OH
90 OH

O
80 -308 (rhamnoglucoside) O OH
Relative Abundance

OH
70
O Glc Rhamnose
60 O Glc Rhamnose OH
OH OH
50 OH OH
OH
-152 (RDA)
40 O+
O HO
O+
30
573.2
20 665.0 881
421.4 545.0 O Glc Rhamnose
10 287.3 O Glc Rhamnose
OH
OH
0
300 400 500 600 700 800 - 308 (rhamnoglucoside)
OH
m/z OH
OH OH

OH O O
OH

O Glc Rhamnose O Glc Rhamnose

OH OH
OH
OH OH
OH

O
O+ O+
HO

OH
OH
OH OH

- 308 (rhamnoglucoside)
OH
OH
OH
OH

OH O
OH O OH O

OH OH
OH
OH
OH OH
OH
- 152 (RDA) OH
OH OH
OH
HO O+ O+
HO
O
O+

OH OH
OH OH
OH
OH

Fig. 3. MS2 and MS3 of peak 1 Cy 3-rutinoside dimer at m/z 1189.

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114 M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115

Table 2
Anthocyanin contents (mg/g fresh weight) in the skin of different fig varieties

Compounds Colar % Granilla % Cuello de % Bursa Siyahi %

Dama (dark
purple)

Cy 3-rutinoside dimer 0.9370.03cjk 2.9 0.5470.04bi 0.6 0.4670.03ai 0.5 0.4470.01ai 0.5
(epi)catechin-(4-8)-Cy 3-glucoside tai 0.0 tai 0.0 0.4870.04ci 0.5 0.3870.02bi 0.4
(epi)catechin-(4-8)-Cy 3-rutinoside 1.2370.04bklm 3.9 0.6270.08ai 0.7 0.5970.12ai 0.6 0.6370.04ai 0.7
Cy 3,5-diglucoside 3.8070.06abn 11.9 5.2371.96bj 6.1 1.9970.20ai 2.2 2.4470.26ai 2.5
(epi)catechin-(4-8)-Cy 3-rutinoside 0.9370.04cjk 2.9 0.6570.06bi 0.8 0.5370.06abi 0.6 0.4470.01ai 0.5
(epi)catechin-(4-8)-Pg 3-rutinoside 0.2470.01i 0.8 nd 0.0 t 0.0 t 0.0
(epi)catechin-(4-8)-Pg 3-rutinoside 0.2070.04i 0.6 nd 0.0 nd 0.0 nd 0.0
Carboxypyrano-Cy 3-rutinoside 1.2670.13dlm 4.0 nda 0.0 0.6370.08ci 0.7 0.4870.06bi 0.5
Cy 3-malonylglycosyl-5-glucoside 1.0470.05bjkl 3.3 1.3970.27ci 1.6 0.6370.10ai 0.7 0.5370.03ai 0.5
Cy 3-glucoside 1.5170.03am 4.7 15.3873.42ck 18.0 10.9870.15bj 12.0 10.3370.76bj 10.7
Cy 3-rutinoside 15.4270.53ao 48.5 57.1670.53bl 66.9 73.3677.90ck 79.9 78.3277.53ck 80.9
Pg 3-glucoside 0.6870.09bj 2.1 0.3470.01ai 0.4 0.2970.05ai 0.3 0.6070.09bi 0.6
Pg 3-rutinoside 3.5270.01dn 11.1 0.4770.06ai 0.5 0.7470.16bi 0.8 1.5570.15ci 1.6
Pn 3-rutinoside nd 0.0 nd 0.0 nd 0.0 nd 0.0
Cy 3-malonylglucoside 1.0370.04ajkl 3.2 3.5170.83bj 4.3 1.1070.07ai 1.2 0.6770.03ai 0.7

Total anthocyanin 31.7970.08a 85.2974.26b 91.7879.39b 96.8176.91b

a,b,c,d
Means values in the same column with different letters are significantly different: LSD (po0.05).
i,j,k,l,m,n,o
Means values in the same row with different letters are significantly different: LSD (po0.05). t: traces; nd: not detected.

Table 3
1
Anthocyanin contents (mg g fresh weight) in the pulp of different fig varieties

Compounds Colar % Granilla % Cuello de % Cuello de % Bursa Siyahi %

Dama (green) Dama (dark
purple)

Cy 3-rutinoside dimer nd 0.0 nd 0.0 nd 0.0 0.0870.01i 0.6 nd 0.0

(Epi)catechin-(4-8)-Cy 3-glucoside 0.1370.05bi 0.9 0.0670.01ai 1.3 0.0370.00ai 2.0 0.0770.00ai 0.5 0.0570.00ai 0.9
(Epi)catechin-(4-8)-Cy 3-rutinoside 0.5970.05dij 4.0 0.0770.01bi 1.5 0.0270.00ai 1.3 0.1670.03ci 1.2 0.1470.04ci 2.5
Cy 3, 5-diglucoside 0.1170.02ai 0.8 0.1470.03ai 3.0 0.0970.02aij 5.9 0.5770.08bi 4.3 0.1570.02ai 2.7
(Epi)catechin-(4-8)-Cy 3-rutinoside 0.1370.03ci 0.9 0.0570.00ai 1.1 0.0270.00ai 1.3 0.0970.00bi 0.7 0.0670.00ai 1.1
(Epi)catechin-(4-8)-Pg 3-rutinoside t 0.0 nd 0.0 nd 0.0 nd 0.0 t 0.0
(Epi)catechin-(4-8)-Pg 3-rutinoside nd 0.0 nd 0.0 nd 0.0 nd 0.0 t 0.0
Carboxypyrano-Cy 3-rutinoside 0.9470.12cj 6.4 0.0970.01ai 1.9 0.0470.00ai 2.6 0.1670.03bi 1.2 0.0770.00ai 1.2
Cy 3-malonylglycosyl-5-glucoside 0.0970.01di 0.6 0.0670.01ci 1.3 0.0270.00ai 1.3 0.0770.00ci 0.5 0.0470.00bi 0.7
Cy 3-glucoside 2.1570.25dk 14.7 0.5470.14bi 11.4 0.1970.02aj 12.5 1.6770.39cj 12.6 0.5170.04bi 9.6
Cy 3-rutinoside 10.1671.21cl 69.5 3.5871.11bj 75.8 1.0470.44ak 68.4 10.1571.62ck 78.6 4.4671.57bj 79.2
Pg 3-glucoside 0.0970.01ci 0.6 0.0170.00bi 0.2 tai 0.0 0.0270.00bi 0.2 0.0170.00bi 0.2
Pg 3-rutinoside 0.0870.00bi 0.5 0.0470.00ai 0.8 0.0270.00ai 1.3 0.0870.01bi 0.6 0.0470.00ai 0.7
Pn 3-rutinoside 0.0670.00ci 0.4 0.0270.00ai 0.4 0.0370.00abi 2.0 0.0370.00bi 0.2 0.0270.00ai 0.4
Cy 3-malonylglucoside 0.0970.00ci 0.6 0.0670.00bci 1.3 0.0270.00ai 1.3 0.0770.00ci 0.5 0.0570.01bi 0.9
Total anthocyanin 14.6271.10c 4.7271.24b 1.5270.40a 13.2271.75c 5.671.56b
a,b,c
Means values in the same column with different letters are significantly different: LSD (po0.05).
i,j,k,l
Means values in the same row with different letters are significantly different: LSD (po0.05). t: traces; nd: not detected.

corresponding to 5.8% of total anthocyanins in the nidin 3-rutinoside) which reached 6.4% in the pulp and 4%
pulp). This was the only variety in which the flavanol– in the skin. In the other varieties its levels were around 2%
pelargonidin condensed pigments could be quantified, in the pulp and 0.5% in the skin, except in Granilla skin
in accordance with its relatively high levels of Pg where this pigment was not found. It is curious to indicate
3-rutinoside. that this pigment was in greater percentage in pulp than in
In Colar variety was also noticeable the relative high skin, even if its absolute concentration were higher in this
proportion of the pyranoanthocyanin (carboxypyranocya- latter.
 ARTICLE IN PRESS
M. Dueñas et al. / Journal of Food Composition and Analysis 21 (2008) 107–115 115

4. Conclusions Giusti, M.M., Rodriguez-Saona, L.E., Griffin, D., Wrolstad, R.E., 1999.
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