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Ficus carica

Article  in  Nutrafoods · July 2010

DOI: 10.1007/BF03223339

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Giovanna Nicotra Angelica Mazzolari

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Research

Ficus carica
Research and development of a dry extract

Giovanna Nicotra*, Silvia Vicentini, Angelica Mazzolari Key words

EPO srl, Milan, Italy Ficus carica
Hydroalcoholic extract
*EPO srl Istituto Farmochimico Fitoterapico Polyphenols
Via Stadera 19 - 20141 Milan, Italy
tel +39.02.895 57.1 - fax +39.02.8955 7490
Luteolin
email gnicotra@eposrl.com - web eposrl.co Quercetin dihydrate

SUMMARY fore not visible externally; in their native habitat, flowers

The common fig or Ficus carica L. is a very prolific fruit
tree, commonly grown in the subtropical globe, whose
leaves have a long traditional use as an antidiabetic agent
in the Mediterranean area and as a hepatoprotector in the
Indian subcontinent. In vitro and in vivo research and pre-
liminary clinical data seem to support the empirical uses.
In these studies, extracts of organic solvents (preclinical
studies) or a decoction (clinical trial) have been used. The
former form is scarcely suitable for human use while the
latter is difficult to be reproduced on an industrial scale.
Therefore, a dry powdered extract obtained from fig
leaves by hydroalcoholic extraction has been developed.
Ethanol is a versatile extraction solvent because, by
changing its ratio with water, several mixtures with differ-
ent affinity for hydrophilic or partially lipophilic bioac-
tives can be prepared. The dry extract has been standard-
ized by developing an innovative HPLC analytical
method and the potential actives (total flavonoids and
quercetin dihydrate) have been identified and quantified.
are pollinated by a very specialized wasp, called Bla-
stophaga psenes, which enters the fruit by a small orifice,
called ostiole, situated at the apex of the syconium (1);
the pollinated syconium develops then into a false fruit,
containing a sweet, jelly pulp and the real, tiny fruits,
some achenes, incorrectly called seeds (3).
Beside its edible uses, the fig fruit has been used in tradi-
tional medicine as a laxative, due to the high content of
sugars and mucilages (2,3). The pulp has emollient and
antinflammatory properties (2). Leaves are regarded as a
traditional remedy for diabetes in Spain and Southwest-
ern Europe (4); in India the plant, commonly known as
Anjir, has been used as chewed leaves to treat jaundice
(5), and also liver and spleen disorders (6). The latex
obtained from leaves has been used topically to treat
warts, insect bites and thorn pricks (2).
Figure 1 Ficus carica L. Common fig foliage and fruit

INTRODUCTION
Ficus carica L. (fam. Moraceae) is a deciduous shrub or
small tree, native to the eastern Mediterranean area and
to Asia Minor, where it has been domesticated some
7000 years ago during the Neolithic age (1). It’s now
commonly grown in all subtropical countries because of
its edible, very nutrient fruits (Fig 1).

The coriaceous, dark green leaves are palmate and deeply

lobed (3 to 7 lobes) (1); they contain a sap which is irri-
tant to the skin (2). The pear-shaped fruit, green, yellow-
ish, bronze or purple in color, is actually a false fruit,
namely an infructescence called syconium, consisting in
a hollow floral receptacle with multiple ovaries clustered
on the inner wall (1). The tiny, fleshy flowers are there-

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In recent papers (7-9) the presence of polyphenols, par-
ticularly of some flavonoids such as quercetin and lute-
olin, with antioxidant and radical scavenging activity, has
been detected in the leaves of F. carica. Some studies on
animal models (10-15) demonstrated the efficacy of fig
leaves in the treatment of diabetes mellitus type 1, due to
their hypoglycemic and hypolipidemic properties.
In a pilot clinical trial (16), a decoction of fig leaves,
which is the traditional galenic form, has been adminis-
tered as a supplement to breakfast to 10 patients (6 men,
4 women, aged 22-38 years) with type 1 diabetes melli-
tus. Subjects were randomly divided into 2 groups (n=5);
during the first month patients were given a decoction of
fig leaves (FC) at the dose of 1 cup daily of 13 g of
leaves, and during the next month a non-sweet commer-
cial tea (TC). The patients were managed with their usual
diabetes diet and twice a day insulin injection. Postpran-
dial glycaemia was lower during supplementation with
FC (156.6 ± 75.9 mg/dL vs commercial tea 293.7 ± 45.0
mg/dL (p<0.001)) without preprandial differences. Also
insulin requirements decreased: average insulin dose was
12 % lower during FC treatment. No adverse effects
were reported.
However, the decoction is difficult to standardize and is
very unstable, both chemically and microbiologically;
hence it is unsuitable for industrial development. As far
as we know, no standardized extract is available on the
market.
The purpose of this study was thus to develop an innova-
tive fig leaves extract and at the same time a new analyti-
cal method to standardize it, on the basis of our long
experience as plant extracts manufacturers and our
knowledge about flavonoids.
MATERIALS AND METHODS
The first step consisted in a careful selection of the fig
leaves; different raw materials were compared coming
from several suppliers and different geographical areas.
The leaves coming from a company producing organic
foods based in Piedmont (Italy) were selected; the leaves
were harvested during spring time in the Varaita valley,
which is located in the uncontaminated environment of
the Cozie Alps, and then air dried on racks in the shade
of a well-ventilated area at room temperature, according
to the traditional process, with a final phase in a drying
oven; thereafter, the leaves were ground to a suitable cut
for extraction.
In a second step, we searched for markers that could be
used to standardize the dried leaves. For this purpose, we
selected two components with antioxidant properties, the
flavonoids luteolin and quercetin, since the presence of
antioxidants has been associated with the normalizing
activity of fig leaves on postprandial hyperlipidemia and
hyperglycemia (10,14).
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Spectrophotometry
The spectrophotometric content of total polyphenols
allows to quantify the rough amount of bioflavonoids in
the sample, which are the natural antioxidants of plants.
In general, there is a direct relation between the
bioflavonoids content and the antioxidant activity of the
sample. Total polyphenols are determined by the Folin-
Ciocalteau method which measures the reducing capacity
of the sample. Gallic acid is used as a reference standard.
Quercetin, was quantified by HPLC.
HPLC
Although we could not quantify luteolin because the ref-
erence standard (luteolin 7-O-glucoside) is highly unsta-
ble, it was possible to develop an innovative method to
quantify quercetin. For this purpose, two different instru-
ments were used, HPLC Waters 2695 (Waters corpora-
tion, Milford, MA-USA) and HPLC Merck La Chrom
(Merck GaA, Darmstadt, Germany), both equipped with
a diode array detector. We developed an HPLC gradient
elution method using a 5μ RP-18 Lichrosphere column
(Merck GaA) and a mobile phase consisting of a water
solution of phosphoric acid and methanol. Quercetin
dihydrate powder as standard (Roth, Karlsruhe, Ger-
many) was dissolved in a mixture of methanol and acid
water whereas, for the preparation of the sample solution,
a hydrolysis reaction in the same solvent was carried out.
Both analyses were carried out in triplicate.
Extraction
After identifying and quantifying the specific markers in
the raw material, the aim was to identify the best extrac-
tion method to produce a dry powdered extract; a mixture
of water and ethanol (extractive menstruum) was used in
the attempt to avoid extraction or purification phases
with organic solvents, as the purpose was to obtain an
extract completely free of harmful residual solvents. Two
different water/ethanol mixtures were evaluated; each
mixture was used to produce three laboratory samples of
the dry extract. The samples were analyzed and the
yields calculated in order to obtain a set of repeatable
results which could allow the choice of the best extrac-
tive menstruum.
New pilot equipment was developed, similar to the one
that will be used in the production plant, but on a small
scale. The pilot equipment is useful to evaluate the feasi-
bility of the extract on an industrial scale and to check the
repeatability of the results previously obtained in the labo-
ratory on larger amounts of raw material. Two batches of
dry hydroalcoholic extracts were produced using the most
suitable ethanol-water menstruum, and the same set of
analysis previously carried out on the laboratory samples
was performed in order to calculate the yield.
The dry extracts were obtained by concentrating and dry-
ing all the percolates; vacuum conditions and tempera-
ture were constantly monitored. The moisture content of
the final extracts was <5%. The extracts were then
reduced to a fine powder by grinding and sieving.
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RESULTS AND DISCUSSION

Figure 2 Total polyphenols and quercetin dihydrate in five 50% hydroalco-
holic extracts (three laboratory and two pilot samples)
The average spectrophotometric polyphenols
content was 1.86%, while the quercetin content Total Polyphenols Quercetin Dihydrate
10.00 0.50
by HPLC was 0.06%, exactly as reported by 9.00 0.45
Vaya and Saeed (7). 8.00 0.40
7.00 0.35
Ethanol-water extraction 6.00 0.30
5.00 0.25
Two different hydroalcoholic mixtures were 4.00 0.20
compared as an extractive menstruum; both had 3.00 0.15
a relatively high content of ethanol (50% and 2.00 0.10
1.00 0.05
70%) in order to solubilize quercetin. Indeed,
0 0
quercetin is soluble in pure ethanol (1 g in 290
mL), in glacial acetic acid, in alkaline solutions, Laboratory extract Laboratory extract
but is insoluble in water.
Pilot extract Pilot extract
The analytical results indicate that both the 50%
and 70%) extraction mixtures (produced a good yield of extraction method is standardized and the technology is
about 60% (Table 1); in any event, the 50% ethanol- suitable to the raw material.
water mixture was selected because the 70% ethanol-
30% water mixture contains a larger amount of chloro-
phyll, which may raise some problems during the drying CONCLUSIONS AND PERSPECTIVES
process.
The leaves of Ficus carica L. contain polyphenols with
Table 1 Content and average yield (w/w) of dry extracts of Ficus
antioxidant and radical scavenging properties which are
carica L. leaves extracted with 50% or 70% of a hydroal- potentially beneficial for human health. Fig leaves have
coholic mixture. Data are the means of triplicate deter- been traditionally used to treat diabetes and liver disor-
minations
ders; but human data supporting traditional uses are still
Ethanol extract (%) scarce. In the only clinical trial available, a traditional not
Parameter 50% 70% standardized galenic form, namely a decoction of the
leaves, had been used. In preclinical studies, extracts
Total dry matter 19.00 15.22 obtained by mean of organic solvents were used. As far
Total Polyphenols 5.63 6.94 as we knew, no standardized extract was available on the
└> extractive yield 57.50 56.80 market, and therefore we developed a new, simple,
Quercetin 0.22 0.25 repeatable extraction method to obtain a dried powdered
└> extractive yield 53.83 63.42 extract of fig leaves, with a fixed content of polyphenols
and quercetin dihydrate, i.e. a standardized extract.

Pilot equipment extraction
The dry extracts of the 50% hydroalcoholic mixture
obtained from the pilot equipment showed a good yield
(more than 60% for quercetin) (Table 2). The data for
each parameter were homogeneous even when compared
to the laboratory samples (Fig 2) indicating that the
On the basis of the available literature describing prelim-
inary preclinical and clinical studies, fig leave extracts
are potentially promising both in the field of food supple-
ments and cosmetics. A new standardized hydroalcoholic
extract is now available for future research.

Table 2 Content and average yield (w/w) of dry extracts of Ficus REFERENCES
carica L. leaves extracted with 50% of a hydroalcoholic
mixture of two pilot exracts. Data are the means of tripli- 1 Morton J (1987)
cate determinations Fruits of warm climate Julia F Morton, Miami (FL-USA), p 47-50
2 Chevallier A (1996)
Parameter Pilot Extract 1 Pilot Extract 2 The Encyclopedia of Medicinal Plants Dorling Kindersley, London
(UK), p 209
Yield (dry matters) 17.02 16.96 3 German Commission E (1990)
Total Polyphenols 5.71 5.49 Monographs Vol 3, Bundesanzeiger N° 101
└> extractive yield 52.25 50.06 4 Yeh GY, Eisenberg AM, Kaptchuck TJ, Phillips RS (2003)
Quercetin 0.23 0.24 Systematic Review of Herbs and Dietary Supplements for Glycemic
└> extractive yield 65.24 70.86 Control in Diabetes
Diabetes Care 26(4) 1277-1294

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