INTRODUCTION

Metabolic syndrome is a syndrome with an aggregation of multiple risk factors consisting of

dyslipidemia, hypertension, diabetes mellitus, obesity and cardiovascular diseases which has become one of the
prominent public health concerns globally. It has got great attention over the past many years. Its significance
lies in the fact that it provides a means of identification for the people who are at high risk of CVDs and Type-2
DM (Alberti et al. 2005).
Metabolic Syndrome is actually a complex interconnected web comprising of physiological, clinical,
pathological, biochemical and metabolic conditions that are a direct cause of increased rate of CVDs and type-2
DM which further contributes in increased rate of morbidity and mortality. Visceral obesity is the cause of
chronic inflammation whereas the production of abnormal adipocytokines (interleukin-1 and 6, adiponectin,
leptin etc.) results in insulin resistance (Kaur 2014).
Obesity and sedentary living style have been considered its root cause. Its prevalence has been found
very high worldwide; mainly because of the increasing rate of obesity in almost all countries of the world. Data
shows that 20% to 30% of all adult population is affected by metabolic syndrome in most countries of the world
with an even greater prevalence in some particular segments of the populations. The risk of CVDs increases
twice while the risk of type-2 diabetes increases five times in the individuals with metabolic syndrome as
compared to those without the syndrome. As it is clustering of many risk factors and increases the risk of mainly
CVDs and type-2 diabetes, its prevention and management has become need of an hour with the priority focus on
combating its root cause i.e., obesity and physical inactivity (Grundy 2008).
Insulin resistance caused by accumulation of adipose tissues and fatty acids in the cells with a
proinflammatory state contribute towards the pathophysiology of metabolic syndrome. Weight loss, increased
physical activity and lifestyle modifications are the key therapeutic drivers while drug therapy can also be helpful
for the risk reduction for diabetes and CVDs (Eckel et al. 2005).
According to the NCEP ATP III definition, metabolic syndrome is present if three or more of the
following five criteria are met: waist circumference over 40 inches (men) or 35 inches (women), blood pressure
over 130/85 mmHg, fasting triglyceride (TG) level over 150 mg/dl, fasting high-density lipoprotein (HDL)
cholesterol level less than 40 mg/dl (men) or 50 mg/dl (women) and fasting blood sugar over 100 mg/dl.
The overall prevalence of metabolic syndrome among the study population was 63.58%, 43.83%, and
69.14% using the World Health Organization (WHO), the National Cholesterol Education Program, Adult
Treatment Panel III (NCEP-ATP III), and the International Diabetes Federation (IDF) criteria, respectively
(Saklayen 2018).
With the high prevalence of all of these metabolic risk factors, the prevalence of metabolic syndrome in
Pakistan according to different definitions is reported to be from 18% to 46%, as compared to the data obtained
from other countries of South Asia.
This syndrome is the result of a cluster of different components including sedentary lifestyle, poor eating
habits, physical inactivity, overnutrition leading to ultimately obesity and abnormal excessive adiposity involving
reproductive disorders, prothrombotic state, nonalcoholic fatty liver disease and proinflammatory state in the
form of other comorbidities. Central underlying pathophysiological mechanisms leading to metabolic syndrome
remain abdominal adiposity and insulin resistance (Cornier et al. 2008).
Various researches have thrown light with evidence on the therapeutic effects of chickpea (Cicer
arietinum L.) used as an intervention therapy against metabolic syndrome. It has been observed that the
beneficial ingredients present in chickpeas show correcting roles against dyslipidemia, insulin resistance, obesity
and cardiovascular diseases besides increasing the nutritional value of the foods with its addition (Aisa et al.
2019).

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 REVIEW OF LITERATURE
Chickpea (Cicer arietinum L.); also called as the ‘garbanzo bean’ or kabuli/white chana is one of
the important and popular pulse crops grown and consumed in the Asian and African countries (Central
and West Asia, South Europe, Ethiopia and North Africa). Fabaceae and Faboideae are the family and
subfamily respectively of this annual legume. Its seeds have been eaten by humans since around 7,000
BC. Among all the legumes, chickpea is ranked third most essential food legume in the world and it is
basically more consumed and grown in the developing countries; accounting for 11.67 million tons
annually standing third in ranking after beans and peas with 25.66 million tons and 11.69 million tons
respectively in the years 2013 to 2017. According to the data, India currently contributes about 70%
production of chickpeas worldwide with 8,832,500 metric tons of mass production in the year 2013.
Australia, Pakistan, Myanmar, Ethiopia, Turkey, Iran, Mexico, Canada and Russia come after India in the
ranking list production wise. Its major characteristic lies in the fact that it contains high content of
protein, B vitamins (riboflavin, niacin, thiamin, folic acid) and the vitamin A precursor b-carotene, fat,
ash and fiber (mostly soluble fiber) and low content of carbohydrates. Rich macronutrients in it include
potassium, calcium, sodium and magnesium and micronutrients are copper, iron and zinc.
Some anti-nutritional factors (like trypsin inhibitors, chymotrypsin inhibitors, alkaloids, tannins,
phytic acid, saponins, and a wide range of polyphenolic compounds including flavanols, flavones,
glycosides flavonoids, etc.) in chickpeas cause hindrance in nutrient utilization but heat treatment
appears to increase the protein content of chickpeas for better intestinal digestion. Chickpeas have fairly
good amount of unsaturated fatty acids (PUFAs) including linoleic and oleic acids (constituting 50-60%
of chickpea fat) having nutritional benefits. Chickpea protein is rich in lysine and arginine. Important
sterols present in chickpea oil includes b-Sitosterol, campesterol and stigmasterol. Naturally present
unique phytochemicals in whole grains of chickpeas adds health benefits like those present in fruits and
vegetables. Chickpeas are also rich in polyphenolic compounds. Chickpea protein digestibility is the
highest and best for human consumption. Among all the food grains, glycemic index of chickpeas is
probably the lowest.
Chickpeas are also regarded as one of the safest food items to consume during pregnancy and it is
also used as galactagogue in lactating mothers. Overall, chickpea contains many beneficial effective
nutrients which makes it potentially significant in providing a number of health benefits in general and
also against metabolic syndromes (CVDs, diabetes, hypercholesteremia, hypertriglyceridemia, HTN,
obesity etc.) in humans. It can be a very inexpensive diet having potent health benefits. In consideration
to all these physiologic benefits provided by chickpeas and in regard to the reduction of risk of many
chronic diseases by consumption of these, chickpeas can be considered as a functional food in addition to
its known facts of protein and fiber content (Jukanti et al. 2012; Singh and Joshi 2016).
Effectiveness of Chickpea against Obesity:
Chickpeas are rich in protein and fiber, both of which aid weight loss. Fiber of chickpeas gives a
feeling of fullness besides keeping a healthy gut while protein has a contribution in causing satiating
hunger. In 2007, a study done on rats demonstrated the effectiveness of dietary chickpeas on visceral
adiposity, dyslipidemia and insulin resistance. Rats were fed a normal-fat diet (NFD), a high-fat diet
(HFD) or a high-fat plus chickpea diet (HFD+CP) for 8 months. The rats fed with chickpea diet showed a
better plasma lipid profile including decreased TGLs and LDL cholesterol. In addition to this, the obese
rats which were given the chickpea treatment reflected remarkably lower leptin levels and improved
insulin resistance. These research findings make chickpeas a functional food ingredient which may be
potentially effective against dyslipidemia and in the prevention of diabetes (Yang et al. 2007).
Obesity and CVDs risk factors including HTN, DM type 2 and dyslipidemia are interlinked.
Increased BMI increases the risk of CVD and DM type 2. Dietary fiber rich foods are known to decrease
the BMI and foods with low GI contribute in reduced insulin levels and faster weight loss than foods with
higher GI. As chickpeas are a food with low GI and high dietary fiber content, they contribute in weight
loss and obesity reduction (Singh and Joshi 2016).
In a review, the nutritional benefits of chickpeas and hummus (chickpea used as a main
ingredient) were seen in regard to weight management, better regulation of glucose and insulin and
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positively affecting some biomarkers of CVDs; while fulfilling the legume recommended servings
among Americans when eaten with vegetables and whole grains. People who consume chickpeas and
hummus have been shown to have relatively higher intakes of nutrients including dietary fiber ,
polyunsaturated fatty acids, vitamin A, vitamin E, vitamin C, folate, magnesium, potassium and iron.
Dietary bioactives such as phytic acid, sterols, tannins, carotenoids, and other polyphenols such as
isoflavones in chickpeas and hummus have extra nutritional benefits in its consumers. Hummus has a
higher Naturally Nutrient Rich (NNR) score than other dips and spreads and this is the reason why
hummus consumers scored higher Healthy Eating Index 2005 (HEI-2005) score (Wallace et al. 2016).
Protective Effect of Chickpea against CVDs:
Among all non-communicable diseases, CVD remains the biggest cause of morality (52%) and
disability worldwide. (Global Altas on CVD prevention and control WHO). Pulses have been seen to
decrease the CVD and diabetes risk due to high fiber content and low glycemic index. Pulses have the
ability of improving lipid profile, blood pressure and better glycemic control and this fact has been
proved by both observational and experimental researches done on adults with and without diabetes, all
of which are major modifiable risk factors of CVD (Lukus et al. 2020).
Several reviews on effectiveness of pulses against cardiovascular diseases risk factors have
shown the positive effect of consumption of these pulses including the chickpeas on heart health. Several
meta-analyses showed that the pulses consumption reduce CVD risk by decreasing total and LDL
cholesterol, increasing satiety, decreasing the excessive accumulation of adipose tissues, improving
glycemic control and by the reduction in blood pressure and inflammation in the body (Padhi and
Ramdath 2017).
Phytochemicals are the bioactive compounds including enzyme inhibitors, saponins,
phytoestrogens, phytohemagglutinins (lectins), oligosaccharides and phenolic compounds which are very
metabolically beneficial to their frequent consumers. Dietary intake of phytochemicals may provide
health benefits, protecting against numerous diseases or disorders, such as coronary heart disease,
diabetes, high blood pressure and inflammation (Bouchenak and Lamri-Senhadji 2013).
Protective Effect of Chickpea against HTN & Diabetes Mellitus:
As chickpea has the lowest GI among all the food grains so this characteristic gives it a potential
for both decreased incidence and severity of type 2 diabetes. Pulses like chickpeas are rich in resistant
starch and amylose which play role in decreasing the bioavailability of glucose which in turn decreases
the glycemic index (GI) and insulinemic postprandial response. The fairly good amount of soluble fiber
has significant beneficial effects in lowering the risk of atherosclerotic coronary artery disease and delays
the postprandial rise in blood glucose, improves glucose tolerance, decreases insulin requirements and
enhanced peripheral tissue insulin sensitivity thus improving glycemic control.
Seeds of chickpeas provide a cheap source of dietary fiber and bioactive compounds including
phytosterols, saponins and oligosaccharides and they play a role in decreased incidence of type 2 DM,
HTN and coronary heart diseases. One of the dominant PUFAs i.e., linoleic acid in chickpeas gives
nutritional benefits by the production of prostaglandins which have blood pressure lowering effects and
plays role in the reduction of smooth muscle constriction. Chickpeas are rich in folic acid which reduces
the risk of CHD by decreasing serum homocysteine levels. Research supports that high content of fiber in
chickpeas decrease total plasma cholesterol levels in obese subjects and one study has reported its
hypolipidemia and positive effects on insulin sensitivity in rats (Singh and Joshi 2016).
Another study explains the mechanisms in detail by which chickpeas plays role in decreasing
insulin resistance which is a direct cause of T2DM. it involves its antioxidant activity, reduction in
adiposity and increasing of SCFA (produce beneficial bacteria) in the gut etc. (Kaur et al. 2019).
Another detailed research including both animal and human trials done on the plants of family
Fabaceae illustrates their antihypercholesterolemic and antihypertensive effects when regularly consumed
(Arnoldi et al. 2015).
One recent research supports the blood pressure lowering effect of several bioactive compounds
present in protein rich pulses. This antihypertensive effect is achieved as these bioactive compounds
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exhibit Angiotensin-Converting enzyme inhibitory activity which can be further improved by
bioprocessing including germination and fermentation by breakdown of proteins and release of bioactive
compounds and polyphenols. These bioactive compounds and polyphenols also show preventive effects
against cancer, diabetes, microbes and oxidative stress (Maleki and Razavi 2020).
Research highly supports the antidiabetic effect by regular consumption of pulses with evidence.
It includes weight management, better satiety, reducing the risk of obesity, better glycemic control,
decrease in blood lipids and better post prandial blood glucose response thus contributing in overall
prevention and management of diabetes (Ramdath et al. 2016).
Another randomized cross-over trial represents the improved glycemic response and reduced post
prandial glucose of meals having white rice with black beans or chickpeas added due to their low
glycemic index (Winham et al. 2017).
Protective Effect of Chickpea against Dyslipidemia:
To see whether chickpeas lower the lipoprotein level, a study was conducted on 47 adults. It was
a dietary intervention with crossover weight maintenance and a randomized study design. Two dietary
periods were involved in the study i.e., chickpea supplemented diet and wheat supplemented diet and
each diet was given for a duration of at least 5 weeks.  The results showed that chickpea diet significantly
lowered the total serum cholesterol and LDL cholesterol levels wheat-supplemented diet (Pittaway et al.
2006).
In another study done on rats, it was observed that germinated chickpeas given in dietary
supplementation mad the lipid concentrations normal in both serum and liver in the rats which were
increased after ovariectomy (Harini et al. 2015).
Another study showed the significant antihyperlipidemic and antitumor effects of chickpea
albumin hydrolysate (CAH) that it significantly decreases the total serum cholesterol, TGs, LDL
cholesterol and increases HDL cholesterol. Also, increases the rate of tumor inhibition and decreases
tumor volume (Xue et al. 2012).
In another study, it was seen that rats showed improvements in overall lipid profile and lipid
metabolism when chickpeas were included in their diet i.e., significant reductions in total cholesterol,
triacylglycerols, LDL cholesterol and VLDL were observed (Zulet and Martinez 1995).
Study states that saponins found in chickpeas are also biologically significant by reducing plasma
cholesterol by 16-24%. Saponins bind the dietary cholesterol or bile acids and in this way increase their
fecal excretion. In addition to this, βsitosterol which is one of the dominant phytosterols in chickpeas has
a role in incidence reduction of serum cholesterol levels and heart diseases (Singh and Joshi 2016).
Another study done on mice showed positive effects of CPe-3 on lipid profile and dyslipidemia
by blocking the cholesterol transport. It also showed the increased activity of serum superoxide dismutase
and fat excretion by feces (Xue et al. 2018).

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 STATEMENT OF PROBLEM
Metabolic syndrome is a cluster of serious risk factors including hypertension, dyslipidemia, diabetes
mellitus, obesity and cardiovascular diseases. The rate of all these non-communicable diseases is already
very high in a developing country like Pakistan. According to WHO, 58% deaths are caused by NCDs in
year 2016. Obesity and sedentary lifestyle have been considered as the major cause of metabolic
syndrome therefore its prevention has become need of an hour. Due to modernization, people have
become more physically inactive and adopted poor eating habits which leads to obesity and other risk
factors ultimately increasing the risk of developing metabolic syndrome. Therefore, this study is
specifically designed in an attempt to develop an economical, nutritious and effective product that not
only shows therapeutic effect against metabolic syndrome but also matches the taste and need of modern
world. Chickpeas have shown various health benefits against metabolic syndrome and also very easily
available throughout Pakistan.
Objectives of the study are:
1. To check the therapeutic effect of chickpeas against metabolic syndrome
2. Development of chickpea supplemented cookies

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 MATERIALS AND METHODS
The methodology will comprise of 2 following phases:
 Phase-1: Physicochemical Analysis, Product Development & Sensory Evaluation

 Phase-2: Human Trial & Biochemical Analyses (Lipid profile, Cholesterol, LFTs, RFTs, ESR,
BMI, Dietary analysis)

Phase-1: Physicochemical Analysis:

Before initiating this phase, pure sample will be collected from Chaudhary foods private limited
Lahore. Then, sample’s physicochemical analysis (including both proximate analyses and determination
of functional properties) will be done at least three times and the methods followed will all be according
to the methods mentioned in AOAC (2000). It will consist of:
 Total Moisture
 Total Ash
 Crude fiber
 Total Protein
 Total Fat
 Water absorbing capacity
 Oil absorbing capacity

Determination of Moisture Content:

The total moisture content of the chickpea flour sample will be determined by oven drying
method as mentioned in AOAC (2000); sub component 925.09.
wt . of fresh sample−wt . of dried sample(g)
Moisture content (%) = × 100
wt . of fresh sample (g)

Determination of Crude Protein:

Kjeldahl method involving digestion, distillation and titration will be used for the determination
of crude protein according to AOAC (2000); sub component 979.09.
Vol. Of 0.1 N H 2 S O4 ×Vol . Of dilution×0.0014
N (%) = ×100
Wt . Of sample ( g ) × Vol. Of dillution ( mL )
Crude protein = N (%) × 6.25
Determination of Crude Fat:
In order to determine the crude fat content of chickpea flour sample, AOAC (2000) method under
sub component 920.39 (Soxhlet extraction) will be followed. The amount of fat will be calculated from
the difference between the weights of extraction flask before and after extraction process in the form of
percentage.
weight of timple before drying−weight of timble after drying
Crude fat (%) = × 100
weight of sample

6
Determination of Crude Fiber:
Using AOAC (2000); sub component 962.09, crude fiber content of chickpea flour samples will
be calculated including the steps of digestion, filtration, washing, drying and combustion.

wt . of crucible with sample before ashing ( g )−wt .of crucible with ash after ashing(g)
×100
wt . of sample(g)
Determination of Total Ash Content:
For the determination of total ash content in the chickpea flour sample, AOAC (2000) sub
component 923.03 will be used. Weight of total ash will be calculated by difference in the before and
after weights and will be represented in percentage.
( wt . of ash ( g ) +Crucible ) −wt . of crucible
Total Ash (%) = ×100
wt . of sample ( g)
Carbohydrates will be determined by subtracting total moisture, ash, crude protein, fiber and fat from
100.
=100−(Total ash %+ crude fat % +crude protein %+ crude fiber %)
Oil Absorption Capacity:
Method of (Sosulski et al. 1976) will be used to determine the oil absorption capacity of chickpea
flour in which one gram of flour sample will be mixed with 10 ml of soybean oil (specific
gravity=0.9092). The mixture made will be then left for about 30 minutes at room temperature (30-32 C).
Then it will be centrifuged at 300 rpm or 2000 × g for 30 minutes.
Water Absorption Capacity:
To determine the water absorption capacity of chickpea flour sample, (Sosulski et al. 1976)
method will be followed. A mixture of one gram of sample with 10 ml of distilled water will be made and
it will then be allowed to stand at optimum room temperature (30-32 C) for about 30 minutes, will be
centrifuged for 30 minutes at 200 rpm or 2000 × g and will be expressed as percent water bound per gram
flour.
Product Development & Sensory Evaluation:
This step will be consisting of two parts. In the first part, product will be developed followed by
the second part of its sensory and physical evaluation.
Chickpea Supplemented Cookies Preparation:
In order to prepare chickpea supplemented cookies, the required materials will be purchased from
the local market. Chickpea flour will be prepared as described earlier while the flow chart of cookie
preparation is shown in the figure below:
Preparation of raw ingredients

Mixed flour and seived together

Mixed fat and sugar

Dry ingredients added into the cream

7
 Kneaded to make dough

Rolled the dough and cookies were cut

Preheat oven at 1500C for 15-20 min

Bake for 15-30 min at 1300C

Cookies cooled for 5-10 min and packed

Grouping:
Four groups will be made with different percentages of white and chickpea flour:
T-1: In the first group the percentage of white flour and chickpea flour will be 85% and 15%
respectively.
T-2: This group will be having 70% white flour and 30% chickpea flour.
T-3: 55% white flour and 45% chickpea flour will be used in the third group.
T-4: Fourth and the last group will be consisting of 40% white flour and 60% chickpea flour.

Group White Flour Chickpea Flour

s

T-1 85% 15%

T-2 70% 30%

T-3 55% 45%

T-4 40% 60%

Sensory Evaluation:
After sensory evaluation, the best one selected will be used to conduct the clinical based human
trial. Sensory evaluation of cookies will be done using a nine-point hedonic test based on the color,
flavor, taste, texture, and overall acceptability from 15 untrained panelists (Hussain et al. 2006).

Determination of Physical Properties of Final Approved Product:

After sensory evaluation, determination of physical properties of the best and the selected
chickpea supplemented cookies will be done and this will be including:
 Weight
 Height
 Diameter
 Spread Ratio

8
Determination of physical properties will be done by random selection of the cookies.
Weight:
Weight of the cookies will be determined by using an analytical balance.
Height:
Height of the cookies will be determined using a vernier calipers in mm.

Diameter:
Diameter will also be taken with a vernier calipers. Average of two readings will be taken (one
after baking and one after rotating them at 90 C) divided by four. This will represent the final diameter of
the cookies in mm.
Spread Ratio:
Spread ratio will be calculated by dividing diameter/width over height/thickness (in mm).
Phase-2: Human trial and Biochemical Analyses:
In the second and last phase, the best one selected after sensory evaluation will be used to
conduct human based clinical trial and the criteria followed will be as follows:
Sample Size: 25-30 patients will be enrolled for the research study.
Product will be given to the patients for 40 days.
Patients will be asked to take biscuits thrice a day.
Dose Calculation: According to the body weight, dose will be calculated in grams.
Shelf life of cookies: 15-20 days so food will be given for 15 days.
Biochemical Analyses:
Biochemical Analyses will include the following blood tests:
 Lipid profile
 Cholesterol
 LFT
 RFTs
 ESR
Biochemical analyses will be done using commercially available kits and microplate reader equipment in
diagnostic facility of Sheikh Zayed hospital.
Assessment:
Assessment of the patients will be done on the basis of:
 BMI
 Dietary assessment/analysis
Sample Collection:
Samples will be collected three times. First sample will be baseline, second will be at 20 th day and
third sample will be taken at 40th day.
3 Samples taken will be compared.
Inclusion Criteria:
9
 Patients with:
 Diabetes Mellitus (History not greater than 5-7 years), Type-2 only
 Dyslipidemia
 BMI (Overweight)
Consent: Patients falling in our inclusion criteria will be counselled and motivated and then they will
be asked to sign a consent form. After that they will be enrolled for the study.
 Patients will be taking their normal diet but a standard DIET PLAN will also be given them
to follow.
 Counseling and training sessions will be conducted for them to follow the diet properly.
 Only interested patients having health concerns will be enrolled in the study who will then be
determined to follow the diet plan incorporated with chickpea supplemented cookies for the
best results.
 Sensory evaluation and physicochemical analyses will be done in UVAS laboratory.
 Human trial will be conducted in Sheikh Zayed hospital Lahore as discussed with Sheikh
Zayed management & co-supervised by Dr. Atif (M.B.B.S, FCPS), Department of General
Medicine.

Statistical Analysis:
Data will be analyzed by using SPSS software version 23, and results will be obtained by
applying one-way ANOVA at the significance level of < 0.05.

10
 SUMMARY
Background: The prevalence of metabolic syndrome is increasing with the increase in urbanization and
modern lifestyle. People in this modern era have switched to sedentary living style and poor eating habits
including fast foods, processed and packaged food. These unhealthy living standards and eating
behaviors are the leading cause of increased rate of non-communicable diseases such as diabetes, obesity,
hypertension, dyslipidemia and cardiovascular diseases, all these being the cumulative risk factors for
developing metabolic syndrome. The prevalence of metabolic syndrome has been seen as high as 46%
which represents an alarming situation. Obesity and sedentary lifestyle need to be managed as the first
priority. Thus, therapeutic effect of chickpeas supplemented cookies will be an effective, nutritious and
economical alternative source against metabolic syndrome while also meeting the desire of people in this
modern era.
Hypothesis: Chickpea supplemented cookies have a therapeutic effect against metabolic syndrome.
Experimental Design: Commercially available fresh chickpea flour sample will be obtained from
Chaudhary foods private limited Lahore and physicochemical analyses including both proximate analyses
and functional properties determination will be performed in UVAS laboratory. Then chickpea
supplemented cookies will be developed and their sensory and physical evaluation will be done in UVAS
laboratory. In the last phase, human trial will be conducted according to the specific criteria on the
patients of metabolic syndrome and in order to check the therapeutic effect, their biochemical analyses
will be done in Sheikh Zayed hospital Lahore.
Statistical Analyses: Results will be analyzed by applying one-way ANOVA at the significance level of
<0.05, on SPSS software version 23.0.

Expected Outcome: Chickpea supplemented cookies will prove to be helpful in the reduction of insulin
levels, dyslipidemia and body fats.

11
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