For centuries, extracts from plants have been used as folk remedies against various health problems . One of the important plants and herbal medicines is eggplant plant, which has been studied extensively, but its cones wasn’t studied thoroughly until now. This study is concerned with the detection and calibration of phenols in the eggplant cones. All the necessary reagents for detection of tannins, flavonoids and chlorogenic acid were positive, and then calibrated phenols with various solvent showed that the ethanolic aqueous extract has the highest percentage of phenol, followed by the aqueous then methanolic extracts. The properties of this extract in terms of its effects on the inhibition of Leishmania parasites were studied. The ethanolic aqueous extract showed a high measure in the rate of inhibition and is equivalent to IC50=39 μg/ml in vitro, the extract was formulated with cream and applied to randomly treated leishmaniasis patients in a double-blind study. The cases were studied in three groups without injection - with injection of glucantime – with liquid nitrogen treatment. Most of the cases have shown improvement or acceleration of improvement since the first week of treatment, confirming the effectiveness and importance of this plant section, which is usually wasted, that is eggplant cones as a new and undiscovered drug with a prospect to treat leishmaniasis.
Leishmaniasis is a tropical and subtropical disease caused by an intracellular parasite transmitted to humans by the bite of a sand fly, mainly Phlebotomus and Lutzomyia (Europe, Northern Africa, the Middle East, Asia, and part of South America); exceptionally, transmission has also been reported as a laboratory accident. According to the World Health Organization (WHO), leishmaniasis is one of the seven most important tropical diseases and it represents a serious world health problem that presents a broad spectrum of clinical manifestations with a potentially fatal outcome [3,4]. It is found in all continents except Oceania [3,5] and is endemic in circumscribed geographic areas in North eastern Africa, Southern Europe, the Middle East, South eastern Mexico, and Central and South America.
Leishmaniasis, which is the third in vector-borne parasitic diseases, after malaria and trypanosomiasis, it is of great importance in the world due to the treatment and control difficulties . According to World Health Organization (WHO) data, approximately 20 million people in 98 countries around the world are infected and 350 million people are at risk .The parasite is found in amastigote form in the blood and tissues of the vertebrate hosts, and in the promastigote form in the vector sand fly which are invertebrate hosts. Amastigotes are oval or
round shaped, 2-4 μm in size and immobile in macrophage cells of vertebrate hosts. When the parasite multiplies in the cell it destroys the cell, and it can be seen individually or in clusters outside the cell . Promastigote form can be detected in the digestive system of the vector. They are single whip shaped or shuttle shaped with a length of 10-20 μm, a width of 1.5-2.5 μm. Promastigotes proliferate in axenic cultures and in the digestive tract of the invertebrate vector .
Cutaneous leishmaniasis is a zoonotic disease caused by protozoa of the genus Leishmania and the genus Phlebotomus. There are two important stages in the life cycle of Leishmania: amastigotes found in humans and other non-human reservoir mammals, and promastigotes found in sand fly. In the vertebrate host the clinical outcome is depending on the parasite strain and the host immune response. The lesions are confined to the skin and to the mucous membrane. A granulomatous response occurs and a necrotic ulcer forms at the bite site. Macrophages containing amastigotes, which may be killed by sensitized lymphocytes were detected in microscopic smear. The lesion may become chronic, usually accompanied by secondary bacterial infection . It has been treated by a chemical and herbal ways and those treatment give some side effects.
This study highlighted a new plant extract from eggplant calyx
that has proven effective against leishmaniosis in vitro by achieving
IC50 =39 μg/ml and the detection of some active substances in
this wasted part of this plant, Eggplant (Solanum melongena L),
which is a vegetable crop, economically important, consumed
widely in Syria. It is native in South East Asian region and was
domesticated over 4000 years ago . Many studies have proved
the effectiveness of this fruit in the treatment of many diseases
such as diabetes, gonorrhoea, cholera, bronchitis, dyspnea,
dysentery, debilitation and the treatment of hemorrhoids [12,13].
thus, this extract were either completely effective treatment or
effective adjuvant treatment.
Eggplants (Solanum melongena) were purchased from local
markets in Damascus. The fruit calyx were obtained, dried, and
stored away from moisture. Three types of extracts (aqueous
ethanol, aqueous, and methanol) were prepared.
a) Methanolic extract : The methanolic extract was
prepared with a Soxhlet extractor. 30 g of the plant sample
was extracted by 250 ml of 99% methanol for four hours,
after which the extract was collected and dried using a rotary
b) Ethanolic- Aqueous Abstract : The ethanolic-
Aqueous extract was prepared by a Soxhlet extractor. 30g
of the plant sample was extracted with 300 ml of (ethanol-
Aqueous) for four hours, after which the extracts were
collected and dried using a rotary evaporator.
c) Aqueous extract : 30 g of cone powder is placed in
the extraction flask with 200 ml of distilled water and heated
under an ascending cooler for 1 hour and the transcript
is filtered and evaporated using a rotary evaporator until
The Folin-Siocaltolin Tp method uses phenols in tungsten
phosphorus molybdate acid in an alkaline medium resulting in a
blue solution measured at absorption at a wavelength of 760 nm
where a series of the reactions occur by electron transfer of two
phenols leading to the formation of blue complexes.  Prepare
the calibration chain: 0.5 g of gallic acid is dissolved in 10 ml
ethanol → supplemented with water up to 100 ml in a calibration
balloon, preferably kept in the refrigerator for up to two weeks.
Add 0/1/2/3/4/5 and 10 ml of the above phenol solution
to a 100 ml calibration balloon and complete the volume with
distilled water. Then extend the volume with water. We will have
the following concentrations of phenol: 0/50/100/150/250/500
mg / L Gallic acid. We put 20 μl in separate covets and add 1.58
ml distilled water + 100 μL foline reagent and mix well then leave
8 minutes and 30 seconds, then add 300 μL of sodium carbonate
and mix well and leave the pair for two hours at 20 degrees
Celsius. Absorption is measured of wavelength 760 nm versus
bulk (distilled water instead of sample) .
The experiment was designed using a cell culture plate
comprising 96 wells where 500,000 parasites were taken from the
homeostasis stage in each well within a final volume of 100 RPMI
culture medium supported by 10% of the FBS gene. The samples
were immunized in a cooled incubator at 26°C. After 48 hours,
the parasite viability was tested using MTT approved viability
test by treating all wells with a final concentration of 10% of the
pigment for 3 hours. At the end of the incubation time, formazan
crystals formed with a special solvent (MTT Solvent) The samples
absorption was 540 nm wavelength using a microplate reader.
The average percentage of viability of each sample is calculated
using the following equation:
The half maximal inhibitory concentration of the parasite cell
growth IC50 is calculated using the Excel program and plotted the
viability curve in terms of the absorbance of all samples by the
curve equation we calculate the value of IC50 given the viability
values as averages ±standard deviation .
This study was a randomized double-blind, included 30
patients. the treatment was applied as three cases (without
injection - with glucantime injection - with liquid nitrogen) to
evaluate synergistic and non-synergistic therapeutic effects. The
patients were randomly assigned from the dermatology hospital
in Damascus and were provided with the necessary materials
from the Faculty of Pharmacy, Damascus University.
The cream containing the herbal extracts was distributed
randomly and everyone was asked to use it twice a day throughout
the 4-week treatment period. The patients were asked to attend
at the hospital for local injection of the glucantime in the lesion,
and that was only for severe cases requiring injection. Patients
were diagnosed by dermatologists at the Dermatology Hospital in
Damascus university by testing the swab of the lesion to confirm
laboratory infection by detecting the presence of the parasite. The area of infection was determined by centimetre (1-2-3-4) cm and
assessed its change during the treatment period. Both the degree
of redness and edema, as well as the severity of sclerosis and the
severity of secondary ulcers, if any, were determined using the
Likert encoding scale. The clinical observation was weekly.
By comparing our study with Tiwari et al.  study tannins
were consistent with our study of iron chlorine with the difference
that the reaction with lead acetate was positive in the our present
study, and in the interactions of flavonoids Tiwari et al.  (Table
1). It was limited to reagent Shinoda only, in this present study
we had five reactions that all were positive with Chlorogenic acid.
Similar characters indicate that the statistical differences
are not significant while the different letters indicate that (Table
2) the statistical differences are significant, and the value of P
<0.05 was used to indicate the statistically significant difference.
Statistical differences between the values of these extracts is
statistically significant. The aqueous ethanolic extract shows the
highest phenolic content followed by the aqueous extract then the
methanolic extract (Figure 1) (Table 3). By comparing between
current study and previous studies we find that total phenolic
content of the aqueous extract is (1025 mg /100g) and this value
within the specified range which was studied in 33 species of
eggplant of different origins and its value was ranges from (740
to 1430 mg gallic acid 100/g) José et al. , but this study was
on fruit and not on cones. In another study of total phenolic
content in the cones of eggplant Diab et al. [19,21-24] it showed
a significant difference in value which was greater (2869 mg /
100 g), and the probability of difference is due to the extraction
method as the method of preparation was soaking with stirring
for two days. The phenolic content of the ethanolic extract in our
study was (1180 ± 12.83 mg / 100 g), and in the study Diab et al.
 the result was (826 mg /100g), meaning that the ratio was
higher in the extract of this present study, with the difference in
the method of preparation (Table 4 & 5).
In this present study, the eggplant cones contain phenolic
substances, useful in the treatment of many diseases, where the
results of the calibration of phenolic substances showed that
the ethanolic-aqueous extract has the highest phenolic content
followed by the aqueous extract. Each of these components has
its own therapeutic effect as Flavonoids so it play a good role
in treating damaged and inflamed tissues  strengthening
the walls of capillary veins, anti-inflammatory, anti-fungal,
antimicrobial, antibacterial [26,27], antiviral, anti-cancer, antivascular,
anti-allergic, anti-oxidant  preventing injuries caused
by free radicals by Inhibiting the free radical enzyme, activation
and regeneration of antioxidant systems and has role in reducing
the permeability and fragility of the veins  And The rutin
witch founds in the extract is one of the most important types of
flavonoids because it reduces the capillary permeability of blood
vessels . Tannins have the strongest effect among phenols for
their role in strengthening soft tissues, reducing excess secretions
[31-33]and repairing damaged tissues .
It works at the same time to stop bleeding due to its
astringent effect in addition to its antiseptic effect  and it
has an important role in the protection of inflamed surfaces of
human body . It is also considered anti-tumour because it
inhibits the proliferation of cancer cells and prevents DNA from
vandalism [31,37] .In addition to these properties, tannins have
great antioxidant capabilities due to their Phenol nuclei. Most of
the antioxidant effects of phenolic compounds are due to its own
Oxidation Reduction properties, which make them Reduction
agents. Flavonoids and phenolic acids are among the most potent
antioxidant compounds. Phenolic acids also have interesting
biological properties such as Anti-inflammatory and antipyretic
, One of these phenolic compounds is chlorogenic acid it has
a role as antioxidant [31,33], protect the inflamed surfaces of the
mucous membranes, an anti-tumour that inhibits the proliferation
of cancer cells and protects the DNA from destruction .
With these auxiliary effects of phenols in skin restoration and
strengthening of blood vessels, they support the inhibitory effect
of leishmaniasis parasites, especially in the ethanolic-aqueous
extract. where the correlation in the results was corresponds in
terms of the highest rate of phenols and the highest inhibition of
leishmaniasis parasites compared with other extracts (Figure 3
The efficacy in inhibiting the growth of leishmaniasis
parasites in the synthetic medium of ethanol-aqueous extract is
very close to that of glucantime, the drug traditionally used for the
treatment of leishmaniasis. IC50 for the extract =39μg/ml-IC50
for glucantime=35μg/ml . This is consistent with the clinical
results of the cream use containing the extract as the healing of
cases that require liquid nitrogen and glucantime injection was
faster by using the cream and in the case of using it alone the
amount of edema gradually reduced and the severity of infection
and sepsis significantly lessened from the first week and pictures
We can say that ethanolic- aqueous extract of eggplant
cones(calyx), which is neglected in medical and nutritional use
showed a high inhibitory effect of Leishmania parasites close in
its efficiency to the glucatiem effect. In addition to the presence
of active substances in it, including high percentage of phenols,
as it helps to repair the affected area and Accelerates its recovery.
therefore, the use an effective cream to treat leishmaniasis
disease will benefit the patient all over the world which is more
economical, easier to apply, without side effect and less painful in
the treatment, so this is the goal that we seek to achieve.
Vargas-Martínez F, Torres-Guerrero E, Quintanilla-Cedillo MR (2013) Leishmaniasis en Mé Academia Mexicana de Dermatología, Colegio de Dermatólogos de Yucatán A. C., Fundación Mexicana para la Dermatología, Universidad Autónoma de Campeche y Secretaría de Salud. México.
Bello SO, Muhammed BY, Gammaniel KS, Abdu-Ayugel KS, Ahmed H (2005) Preliminary evaluation of the toxicity and some Pharmacological properties of Aqueous crude extract of Solanum melongena. Research Journal of Agricultural and Biological Sciences 1(3): 1-9.
Gill LS (1992) Ethno medical uses of plants in Nigeria. University of Benin Benin, Nigeria, pp. 215.
Baris O, Guluce M, Sahim F, Ozer H, Kilic H (2006) Biological activities of essential oil and methanol exctract of Achillea biebersteinii Afan (Asteraceae). Turk J Biol 1(30): 65-73.
R Diab, A Mounayar, E Maubuf, R Chahia (2011) Benefical effects of solanum melongena (solanaceae) peduncles extracts, in periodontal diseases. Laboratory of oxidative stress, faculty of medicine and dentistry, Lebanese university, beirut, Lebanon, p. 5-11.
Muthsamy P, Sundram N, Desi V, Lakskmanam P (2009) The biochemical basis of shoot and fruit borer resistance in interspecific progenies of brinjal (Solanum melongena). Tamilnadu agricultural university 3: 50-57.
Gopalan C, Rama Sastri BV, Balasubramanian S (2007) Nutritive Value of Indian Foods. published by National Institute of Nutrition (NIN). ICMR.
Benhammou N (2012) Activité antioxydante des extraits des composes phénoliques de dix plantes médicinales de l’Ouest et du Sud-Ouest Algé Thése doctorat. Université Aboubakr Belkaïd.Tlemcen, pp. 174.
Harkat H (2008) Hétérocycles oxygénés et composés aromatiques de Frankenia thymifolia Desf: formation d’hétérocycles oxygénés et isolement de substances naturelles. Thèse de doctorat. Université El Hadj Lakhder Batna, pp. 222.
Athamena S (2009) Etude quantitative flavonoides des grains de Cuminum cyminum et Les feuiles de Rosmarinus officinalis et l’evaluation de l’activite biologique. Memoire Présenté pour l’obtention du diplôme de Magister. Université El -Hadj Lakhder Batna, pp. 126.
Benhammou N (2012)- Activité antioxydante des extraits des composes phénoliques de dix plantes médicinales de l’Ouest et du Sud-Ouest Algé Thése doctorat. Université Aboubakr Belkaïd. Tlemcen, pp. 174.
Ferradji A (2011) Activités antioxydante et anti-inflammatoire des extraits alcooliques et aqueux des feuilles et des baies Pistacia lentiscus. Mémoire Présenté Pour l’obtention du Diplôme de magister. Université Ferhat Abbas. Setif.