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Evaluation of hypolipidemic effect of citrus lemon

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Abstract

Herbs have been used for several cardio vascular diseases, such as congestive heart failure, hypertension, angina pectoris, atherosclerosis, arrhythmia, cerebral and venous insufficiency. Atherosclerosis is the primary cause of heart disease and stroke. Hence blood lipid levels are probably the major determinant of the development of cardiovascular disease. Citrus fruit and juices have long been considered a valuable part of a healthy and nutritious diet. It is well established that some of the nutrients in citrus promote health and provide protection against chronic disease. The present study has been specifically designed to investigate the hypolipidemic effects of citrus lemon juice in rabbits after high cholesterol diet for four weeks. The citrus lemon juice (1ml/kg/day) revealed a significant reduction in serum cholesterol, triglycerides; low density lipoprotein levels and resulted in an increase in high density lipoprotein. These results suggest that the hypocholesterolemic effects of citrus lemon juice may be due to its antioxidant effect.
Journal of Basic and Applied Sciences Vol. 6, No. 1, 39-43, 2010 ISSN: 1814-8085
EVALUATION OF HYPOLIPIDEMIC EFFECT OF CITRUS LEMON
Yasmin Khan, Rafeeq Alam Khan*, Syeda Afroz and Afshan Siddiq
Department of Pharmacology, Faculty of Pharmacy
University of Karachi, Karachi, Pakistan
ABSTRACT
Herbs have been used for several cardio vascular diseases, such as congestive heart failure, hypertension, angina
pectoris, atherosclerosis, arrhythmia, cerebral and venous insufficiency. Atherosclerosis is the primary cause of heart
disease and stroke. Hence blood lipid levels are probably the major determinant of the development of cardiovascular
disease. Citrus fruit and juices have long been considered a valuable part of a healthy and nutritious diet. It is well
established that some of the nutrients in citrus promote health and provide protection against chronic disease.
The present study has been specifically designed to investigate the hypolipidemic effects of citrus lemon juice in rabbits
after high cholesterol diet for four weeks. The citrus lemon juice (1ml/kg/day) revealed a significant reduction in serum
cholesterol, triglycerides; low density lipoprotein levels and resulted in an increase in high density lipoprotein. These
results suggest that the hypocholesterolemic effects of citrus lemon juice may be due to its antioxidant effect.
Keywords: Citrus lemon, Hypolipidemic effect.
INTRODUCTION
Cholesterol is an amphipathic sterol, hydrophilic at the
hydroxy end with a hydrophobic hydrocarbon side chain
and thus can be incorporated into membrane bilayers. As
well as being an essential structural component of cell
membrane, cholesterol determines membrane lipid
organization and structural integrity (Goldstein et al.,
2006; Ikonen, 2006). Hypercholesterolemia is
characterized by high levels of cholesterol in the blood .It
is not a disease but metabolic derangement that can
contribute to many disease, most notably cardiovascular
disease. It is closely related to the terms hyperlipidemia
and hyperlipoproteinemia .Elevated cholesterol in the
blood is due to abnormalities in the levels of lipoproteins,
the particles that carry cholesterol in the blood streams.
This may be related to diet, genetic factors (such as LDL
receptor mutations in familial hypercholesterolemia) and
the presence of other diseases such as diabetes and an
under active thyroid (Durrington etal, 2003).
Hypercholesterolemia is silent disorder that is usually
diagnosed after a heart attack or stroke. Cardiovascular
diseases have become a major clinical and public health
problem in south Asian countries specifically India,
Pakistan, Srilanka, Bangladesh and Nepal. This
population carries increased risk even if they migrate to
other countries (Iqbal et al., 2005).The national
cholesterol education program (NCEP) published a
guideline in 1993 for screening and treating
hyperlipidemia (Eric et al., 2002). The NCEP program
includes therapeutic life style changes such as reduced
intakes of saturated fats to < 7% of total calories and <
200 mg/day cholesterol. 25% to 35% daily fat intake
provides low saturated and trans fats to body. Therapeutic
options to reduce LDL also involve the use of plant
steroids, 2gm/day and increased intake of viscous fiber
(soluble fiber) about 10-25 gm/day. Similarly exercise
and life style modification is also beneficial. However
most of the allopathic drugs are toxic and generate free
radicals in the body (Beldeu, 2006). Thus present study
has been specifically designed to investigate the lipid
lowering effect of citrus lemon with minimal side effects.
MATERIALS AND METHODS
Selection of Animals
The study was carried out on 18 healthy white rabbits of
either sex weighing from 1000-1500 gm. Rabbits were
selected as experimental animals since biochemical
changes produced in rabbits are comparatively similar as
observed in humans (Irena et al., 1979). Moreover
sufficient amount of blood samples can easily be
collected, easy to handle and economical.
Animal Groups
Animals were divided into two groups control and treated
each containing 9 animals. Apparent health of these
animals was monitored during the conditioning period
under the laboratory environment for a week before
administration of drug specifically noticing loss of hair,
diarrhea, hematuria, skin ulceration and loss of activity.
They were housed individually in cages, under controlled
condition of temperature 23±2ºC. Diet and water was
provided ad libitum.
Dosing
Animals of both groups were given high cholesterol diet
(HCD) daily for a period of 30 days i.e. 0.125gm/kg
cholesterol (Merck) in 0.5% corn oil purchased from local
*Corresponding author: email: rkhan1959@gmail.com
J. basic appl. sci.
40
market. After 30 days animal of treated group were
administered fresh citrus lemon juice orally for 30 days in
the dose of 1ml/kg during the first phase of study, while
animals of control group were administered water through
same route equivalent to the volume of respective doses
according to their body weight. During the second phase
of study animals of treated group were further
administered fresh citrus lemon juice up to 45 days and
compared with control for the same period.
Sample Collection
Blood samples of 5 ml were collected thrice, first after 30
days of HCD then again after 30 and 45 days dosing of
citrus lemon juice from ear vein of the animals in gel tube
for evaluation of lipid profile.
Gross Toxicities
Overall gross toxicities such as skin ulceration, average
weight variation, hematuria, alopecia, loss of activity,
diarrhea, salivation, aggressive behavior and color of skin
in different groups of animals was observed during the
total period of experiment.
Animals of control group showed no gross toxicities, but
there was an overall increase in average weight of control
animals by 15 to 35 gm however animals at HCD
comparatively showed more increase in average weight of
the animals by 25 to 260 gm during the total period of
experiment except in second week, where there was
decrease in weight by 75 gm. Animals at HCD revealed
slight gross toxicities, such as yellow skin, alopecia
(especially at neck and foot), yellow urine with acidic pH
and loss of activity that may be due to severe diarrhea.
Animals at citrus lemon (1ml/kg) after HCD did not
reveal gross toxicities. However there was loss of weight
of 30 to100 gm during the total period of study. However
at the end of study there was 53 gm decrease in average
weight.
MORTALITY RATE
Table 1 reveals mortality rate of animals kept on HCD for
30 days, treated with lemon and control animals for 45
days. Deaths were observed in animals at HCD during 2nd
and 3rd weeks of dosing. However no death was observed
in animals of control and treated groups.
Table 1. Comparison of Overall Mortality Rate (%)
Animals Groups Mortality Rate
HCD 07/25(28.00)
Control 0/9(00.00)
Lemon 0/9(00.00)
No. of animals expired/total animals (percentage)
BIOCHEMICAL ANALYSIS
Serum samples collected at the end of dosing were
utilized for assessment of lipid profile using standard
reagent kits of Human Germany. All the values were
expressed as the mean and standard error to the mean.
Student test was performed and P values were observed.
Values of P<0.05 were considered as significant and
P<0.005 as highly significant. All statistical procedures
were performed according to the method of Alcaraz and
Jimenez (1989).
RESULTS
Table 2 reveals the comparative effects of citrus lemon on
cholesterol, triglycerides, LDL and HDL at the dose of
1ml/kg citrus lemon for 30 and 45 days respectively in
animals received high cholesterol diet for 30 days.
After 30 days animals showed highly significant decrease
in cholesterol 150.8 ± 19.1mg/dl in comparison to control
animals i.e. 345.3± 28 mg/dl. While after 45 days
decrease in cholesterol continued and a highly significant
decrease was observed i.e.16.9 ± 2.8 mg/dl in comparison
to the value of control animals i.e. 232.6±9.0 mg/dl.
Similarly a highly significant decrease in LDL
concentration was observed after 30 days i.e. 122.6± 13
mg/dl in comparison to control animals i.e. 273 ± 38
mg/dl. The decrease in the levels of cholesterol and LDL
persisted even after 45 days. Where as HDL concentration
was increased significantly after 30 days i.e. 7.27 ± 1.7
mg/dl than as compared to control i.e. 3.07 ± 0.26 mg/dl.
However increase in HDL level became insignificant after
45 days i.e. 2.15 ± 0.31 mg/dl as compared to control i.e.
2.1±0.32 mg/dl.
Decrease in triglycerides was insignificant after 30 days
as compared to control animals. However a highly
significant decrease was observed in triglyceride levels
after 45 days i.e. 26.69 ± 1.4 mg/dl in comparison to
control 40.3±3.5 mg/dl.
Table 3 reveals the %age reduction of cholesterol,
triglycerides, LDL and HDL at 1ml/kg dose of citrus
lemon for 30 and 45 days respectively in animals received
high cholesterol diet for 30 days.
DISCUSSION
Herbs have been used as medical treatments since the
beginning of civilization and some derivatives (e g,
aspirin, reserpine, and digitalis) have become mainstays of
human pharmacotherapy. Herbs have been used for
several cardiovascular diseases, such as congestive heart
failure, hypertension, angina pectoris, atherosclerosis,
arrhythmia and cerebral insufficiency. However, many
herbal remedies currently in use have not undergone
Khan et al. 41
careful pharmacological assessment, and some have the
potential to cause serious toxic effects and major drug-to-
drug interactions. The high prevalence of herbal use in
current medical practice throughout the world has
increased questions about the safety and efficacy of herbal
drugs for cardiac disease and clinicians must be informed
about the potential for benefit and harm. Thus continuous
research is necessary to elucidate the pharmacological
activities of the herbal remedies now being used to treat
cardiovascular diseases (Nick et al., 1998).
Vegetables, fruits and their constituents are potent
effectors of biological system in humans (Lampe,
1990).The cholesterol-lowering effects of vegetables,
fruits and their constituents have been examined in detail.
Daily doses of these food showing cholesterol-lowering
effects in human have been reported as follows; variety of
vegetables (750g /day) and fresh apples (350-400g/day),
guava fruit (0.5-1kg /day) , garlic (10-20g /day) and a low
saturated fat diet combined with vegetables, protein
(33g/day) and a high soluble fiber(18g/day) (Lampe,1990;
Jenkins, 1999).
Atherosclerosis is the primary cause of heart disease and
stroke. Hence, blood lipid content is probably the major
determinant of the development of cardiovascular disease.
Several authors have reported the hypolipidemic effects
of Citrus juice (Monforte, 1995; Gorinstein, 2004)
moreover, some studies suggest that a high dietary intake
of orange or grape juice might reduce hypercholestero-
lemia, and this was postulated to be largely due to the
principal Citrus flavanones, hesperidin from orange and
naringenin from grapefruit, as their glycosides, hesperitin
and naringin (Kurowska, 2000).
0
50
100
150
200
250
300
350
400
Cholesterol LDL HDLTriglycerides
serum mg/dl
0
50
100
150
200
250
300
Cholesterol LDL HDL Triglycerides
serum mg/dl
Fig. 1. Histogram showing comparative effects on lipid
profile in HCD and citrus lemon animals after 30 days. Fig. 2. Histogram showing comparative effects on lipid
profile in HCD and citrus lemon animals after 45 days.
Table 2. Comparative effects of citrus lemon on lipid profile after high cholesterol diet for 30 and 45 days
No. of Days Groups Cholesterol
Mg/dl LDL-CHOL
Mg/dl HDL-CHOL
Mg/dl Triglycerides
Mg/dl
Control 345.30±28.0 273.0±38.0 3.07±0.26 39.40±4.7 30 Lemon 150.80±19.1** 122.6±13.0** 7.27±1.7* 39.40±4.2
Control 232.60±9.0 148.1±19.0 2.10±0.32 40.30±3.5 45 Lemon 16.94±2.80** 76.8±6.10** 2.15±0.31 26.69±1.4**
n = 9, Average ± SEM, *p < 0.05 significant as compared to control, **p < 0.005 highly significant as compared to control
Table 3. Percentage reduction in lipid profile after 30 and 45 days administration of citrus lemon
Cholesterol LDL HDL Triglycerides
No. of days Groups %age
Control 51 52.2 21.4 36.1 30 Lemon 79 80.5 20.8 64
Control 68.5 73.8 40.4 41.2 45 Lemon 97.7 87.8 65 67.1
serum conc. m
g/
dl
serum conc. m
g/
dl
J. basic appl. sci.
42
The present study has been specifically designed to
investigate the hypolipidemic effects of Citrus lemon
juice in rabbits after high cholesterol diet for 4 weeks.
The Citrus lemon contains important natural components,
including citric acid, ascorbic acid, minerals and
flavonoids (Benavente et al, 1997) though health-related
properties of citrus lemon have been associated with their
content of vitamin C, but recently flavonoids have been
also shown to play a role in this respect. Some researchers
suggest that flavonoids have different biological
functions, including antioxidative, anti-inflammatory,
antiallergic, antiviral, antiproliferative, antimutagenic and
anticarcinogenic activities (Middleton & Kandaswami,
1986; Godeberg, 1994, Meyer, 1994 and Rice, 1997).
The citrus lemon juice (1ml/kg/day) revealed a significant
reduction in serum cholesterol, triglycerides, low-density
lipoprotein levels and resulted in an increase in high-
density lipoprotein(Fig 1 & 2). These results suggest that
the hypocholesterolemic effect of citrus lemon juice may
be due to its antioxidant effect since previous studies has
suggested that antioxidant (vitamin C) administration in
hypocholesterolemic rats improves endothelial function of
coronary and peripheral vessels (Henry et al., 1997). It is
widely accepted that elevations in cholesterol and LDL
plasma levels are major factors for coronary heart disease
There has been relationship between elevated LDL and
atherosclerosis. Since LDL in the blood gets deposited in
the blood vessel walls and becomes the major component
of atherosclerotic plaque. Studies suggest that
pathological process could be reversed by reducing the
serum LDL level (Ross, 1993). Since citrus lemon juice
has significantly reduced cholesterol and LDL levels
therefore it may be concluded that citrus lemon might
prevent atherosclerosis. Moreover there has been
significant increase in serum HDL levels which is
considered to be good cholesterol (Stein, 1999). The
increase in HDL has shown to slow down the
atherosclerosis process (Nofer, 2002). Therefore it can be
safely concluded that citrus lemon juice could be
considered as a valuable supplement to prevent coronary
diseases.
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... The guidelines for making the mixture was adopted and modified from the research conducted by Hasan et al. and Khan et al.(Hasan, Singh, Siddiqui, Kulshreshtha, & Aggarwal, 2013;Khan, Khan, Afroz, & Siddiq, 2010). The way to make the mixture is using a lemon (Citrus limon) and virgin olive oil (Olea europea). ...
... The results of the paired sample t-test statistical test proved that there were significant differences (p=0.023) of cholesterol levels in hypertensive patients in Gilangharjo Village, Pandak, Bantul, Yogyakarta before and after the treatment of a mixture of lemon and virgin olive oil. The results of this study support the results of previous studies conducted by Violante et al. (2009) and Khan et al. (2010), which stated that the provision of virgin olive oil and lemon juice could reduce cholesterol levels in hyper cholesterol patients (Violante, Gerbaudo, Borretta, & Tassone, 2009;Yasmin, Khan, Syeda, & Afshan, 2010). However, the results of this study are somewhat different from the results of research conducted by Yuiwarti et al. (2018), which stated that compared to virgin olive oil, virgin coconut oil has more potential to maintain cholesterol in hyperglycemic Rattus rattus norvegicus (Wachidah Yuiwarti, Saraswati, & Kusdiyantini, 2018). ...
... Considering the administration of lemon juice in rats with a high cholesterol diet intake for four weeks. Khan et al.(Khan et al., 2010) showed that taking 1 ml/KgWB of juice or lemon juice/day significantly reduced serum cholesterol levels, triglycerides, LDL, and increasing levels of HDL. In short sentences, lemon reduces the risk of heart disease through the content of eriocitrin and hesperidin, which has antioxidant properties. ...
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The study evaluated the effect of mixtures of lemon and virgin olive oil on the cholesterol level in hypertension patients. A quasi-experimental study, with a pretest-posttest without control group design, did on 18 purposively sampling hypertension patients. Samples received mixtures of 30 ml lemon squeeze and 30 ml virgin olive oil once a day 30 minutes before breakfast for 14 days. The cholesterol level was measured the day before (day 0) and the 14 th days of administration. The pretest's mean cholesterol level was 265.83±53.52 mg/dl, and the posttest was 228.67±55.14 mg/dl. Paired sample T-test showed significant differences before and after administration of mixture (p= 0.023; p<0.05).
... S e v e r a l pharmacological properties have been ascribed to different members of Citrus species. These properties are anticancer (26,27,28,29,30), antifungal (31), antityphoid (29), anti-oxidant (32), antiulcer (33), hypolipidemic (34). Carvone and limonene have a broad range of anti-fungal and anti-microbial activities (35,36). ...
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The objective of this study is to develop a new and effective antibacterial agent against food pathogens that poses a major threat to human health and to investigate the antioxidant activity of this plant. Methanol, ethanol, and aqueous extracts were analyzed for antimicrobial potency. Eight different microorganisms were used in the study, one of which was yeast. These microorganisms are food pathogens. Antimicrobial activity testing was performed using a disc diffusion method. Another test for antimicrobial activity is the minimum inhibitory level. Antioxidant activity was conducted using 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•) and 1,1-diphenyl-2-picrylhydrazyl (DPPH•). Citrus limon ethanol extract had a maximum inhibiting zone against Escherichia coli (12 mm). In the ABTS method, the highest scavenging activity was obtained from an ethanol extract (58.3 percent). The methanol extract (95.4%) led to the largest DPPH scavenging activity. Consequently, Citrus limon extracts have antimicrobial, antioxidant, and anti-biofilm potential against foodborne pathogens.
... Moreover, by supplementing rabbit diets with 1% or 2% DLP showed a significant reduction in serum triglycerides, cholesterol, LDL-c, and VLDL-c were significantly reduced, while the level of HDL-c level statistically increased compared to the level in control animals. The hypolipidemic effect of C. limon may be due to its flavonoids content, which can reduce blood cholesterol, triglycerides, and LDL-c levels based on its antioxidant effect [46]. Complete methoxyylation of ring A with Citrus flavonoids seems to be an optimal structural change that exhibits substantial effects on the modulation of hepatic lipid metabolism via the suppressed secretion of lipoproteins with apoB in HepG2 cells [47]. ...
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The current study examined the influence of Citrus limon (dry lemon) on the hemato-biochemical profiles, and antioxidant indices of growing rabbits. Forty-eight growing New Zealand White rabbits (age, eight weeks; weight, 1543.33 ± 25 g) were allocated into three groups (16 animals each), the first group was (control) fed a basal diet, whereas the second and third groups were supplemented with dried lemon, 1% or 2% DLP, respectively. A GC-MS analysis of more than 27 active constituents was performed. Feed conversion efficiency was (p < 0.05) better with diets containing 1% or 2% dry lemon, compared to the control group. Hematological indexes were increased significantly with the addition of DLP compared to those in the control group. Adding 1% or 2% dry lemon to rabbit diet increased (p < 0.05) enzymatic and non-enzymatic antioxidant activities (TAC, SOD, GSH, GST, and CAT) in serum and liver tissues. Taken together, these data reveal the advantages and antioxidant effects of dry lemon supplementation for growing rabbits once supplemented at a maximum of 2% in their daily diet.
... Moreover, lemon and lemon peels essential oil are reported to be a good source of a wide array of bioactive compounds such as vitamin C, flavonoids, limonoids and folic acid 7 among others. Various studies on Citrus limon suggested that it can also prevent the incidence of cardiovascular diseases 8 . Citrus EOs have been employed as flavoring agents in many products such as cosmetics, foods, medicine and beverages as well are used for aromatherapy. ...
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This research work was carried out to assess the variations in the chemical composition, yield, antibacterial and haemolytic activities of hydro-distilled peel essential oils from unripe and ripened limon [Citrus limon(L.) Osbeck] fruit. The extracted essential oil yield from unripe fruit peel (1.79 %) is slightly lower as compared to the ripened fruit peel oil (1.81 %). By GC/MS analysis, 14 compounds were identified in unripe while 11 components in ripened lemon peel essential oil (LPEO). The major components identified in unripe lemon peel essential oil were caryophyllene oxide (16.791 %) followed by 3-cyclohexene-1-methanol (11.463 %), 2-isoprpenyl-5-metyhlhex-4-enal (10.174 %), limonene (9.649 %) and β-citral (8.323 %) whereas ripened LPEO contained α-Terpineol as major compound (38.497 %) followed by 2,6-octadienal-3,7-dimethyl (9.981 %) and limonene (9.195 %). Both the isolated lemon peel essential oils (LPEOs) exhibited considerable antibacterial activity against E. coli, Bacillus subtilis, Salmonella typhimurium and Staphylococcus aureus with no cytotoxic effects. Overall, a significant variation in the yield, volatiles composition and biological activities of LPEOs were observed that can be linked to lemon fruit ripening stages.
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The use of Essential Oils as antimicrobial agents have become popular over the years in an attempt to find alternative ways of dealing with strains of bacteria that have become resistant to conventional antibiotics. This study was carried out to compare the antimicrobial effects of Citrus peel essential oils obtained from Okene Main Market, 7'33'4.39'' N 6'14'9.20'' E, Kogi State, Nigeria, on the clinical isolates of some microorganisms (Escherichia coli, Pseudesomonas aeruginosa, Staphylococcus aureus, and Aspergillus niger). The oils were extracted from the peels using the cold maceration method with n-hexane as the solvent. The agar diffusion method was used to test the susceptibility of the micro-organism strains using ciprofloxacin as the standard positive control. The experiment was carried out in duplicates and obtained data was analysed using one-way analysis of variance (ANOVA) and Duncan Multiple Range Test (DMRT), with P<0.05 considered significant. The results revealed that Orange (Citrus sinensis) exhibited the inhibitoriest effect on the test isolates followed by lime (Citus aurantifolia) and Lemon (Citrus Limon) with the least significant effect.
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There is currently much interest in phytochemicals as bioactive components of food. The roles of fruit, vegetables and red wine in disease prevention have been attributed, in part, to the antioxidant properties of their constituent polyphenols (vitamins E and C, and the carotenoids). Recent studies have shown that many dietary polyphenolic constituents derived from plants are more effective antioxidants in vitro than vitamins E or C, and thus might contribute significantly to the protective effects in vivo. It is now possible to establish the antioxidant activities of plant-derived flavonoids in the aqueous and lipophilic phases, and to assess the extent to which the total antioxidant potentials of wine and tea can be accounted for by the activities of individual polyphenols.
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Flavonoids are a widely distributed group of polyphenolic compounds with health-related properties, which are based in their antioxidant activity. These properties have been found to include anticancer, antiviral, antiinflammatory activities, effects on capillary fragility, and an ability to inhibit human platelet aggregation. The antioxidant capacity of any flavonoid will be determined by a combination of the O-dihydroxy structure in the B-ring, the 2,3-double bond in conjugation with a 4-oxo function and the presence of both hydroxyl groups in positions 3 and 5. Flavanones, flavones, and flavonols are the flavonoids present in Citrus, and although flavones and flavonols have been found in low concentrations in Citrus tissues, in relationship to flavanones, these types of compounds have been show to be powerful antioxidants and free radical scavengers. Some Citrus flavonoids can be used directly as repellents or toxins or be used in plant improvement programs to obtain more resistant crops. In addition, some Citrus flavonoids and their derivates, in the field of food technology, are principally known for their ability to provide a bitter or sweet taste and as bitterness inhibitor. Keywords: Free radicals, antioxidant; anticarcinogenic; antiinflammatory; platelet aggregation; antiallergic; analgesic; antimicrobial; food additives
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