CHAPTER ONE

Introduction
1.1 Background to the study
Lima bean, Phaseolus lunatus belongs to the family leguminosae and is mostly
cultivated in South America (Allen &Allen, 1981). Lima beans are sometimes referred
to as: haba beans, sugar beans, butter beans, Guffin beans, civet beans, Hibbert beans,
Pallar beans, Sieva beans, Madagascar beans, and Burma beans. It is known as butter
bean in English lima wake in Hausa, lima irisi in Yoruba, the varietal differences exist
in size (small and large) and colour, usually ranging from green to creamy white and a
phenomenal starchy flavor (Mackie, 1983).
Lima bean like all other legumes are food resources that offer various optimum
nutritional and/ or health benefits (Tharanathan & Mahadevamma, 2003). The protein
content of legume grains ranges from 17 to 40 g/100 g, much higher than that in cereals
(7-11.8 g/100 g) and approximately equal to the protein content of meat 18-25 g/100 g
(De Oliveira et al., 2006). Lima beans are rich in the mineral manganese, a key nutrient
in brain development. It’s necessary for the creation of neurotransmitters that help brain
cells communicate, and its antioxidant properties protect against brain cell damage. In
addition to fiber, lima beans are high in plant-based protein. The advantage of lima
beans is that they offer health benefits like a healthier gut and lower risk of heart
disease. Plus, their mild taste makes them super versatile and can be tossed into just
about any dish.
In Nigeria and other developing countries, legumes are the second largest sources of
human food after cereals. Lima beans is the fourth important legume food in Ghana
after cowpea, groundnut and bambara groundnut. The high protein content as well as
the presence of the amino acid lysine makes them a suitable cheap source for
fortification of cereal diets in most economies in transition (Kaur et al., 2009). Health
problems such as hypertension, gall-stone formation which are related to animal protein
consumption have raised great social and public health attention recently. Thus, plant
protein of which legumes form a great majority has been found to play an important
role in several favorable physiological responses, such as reducing heart and kidney
diseases, lowering the sugar indices of diabetic patients, increasing in satiety, and

1
 reducing the occurrence of cancer (De Oliveira et al., 2006). The utilization and
exploitation of a crop generally depends on the available knowledge on it. The dearth of
information on Lima beans in lieu with their optimum nutritional and functional
properties has made this potentially beneficial legume under-utilized. As with legumes,
some undesirable constituents of lima beans include anti-nutrients (trypsin inhibitors,
phytic acid, saponins, haematoglutinns and tannins) that interfere with absorption and
utilization of important minerals such as calcium, iron, zinc and magnesium. Lectins
presents in beans are polymeric proteins that bind to monosaccharide in glycoproteins
of the cell membrane causing lesions in the intestinal mucosa and reduced nutrient
absorption. Lima beans are also known to contain relative high amounts of cyanogenic
glycosides (linamarin or phaseolunatin) and thus need to be treated prior to utilization.
It is known that processing methods such as dehusking, soaking, germination, cooking
and roasting inactivate the antinutrients and can be used to improve the nutritional
quality of the beans (Deshpande et al., 1991).
1.2 Statement of the problem
It refers to the investigation of the specific compounds and nutrients present in lima beans
and their potential health benefits. Lima beans are a type of legume that are rich in
protein, dietary fiber, vitamins, and minerals. They are also a good source of complex
carbohydrates and low in fat. Some of the key nutrients found in lima beans include
folate, iron, potassium, magnesium, and vitamin B6. These nutrients play important roles
in various bodily functions, such as supporting red blood cell production, maintaining
healthy blood pressure, and promoting proper nerve function. Additionally, lima beans
are known to contain phytochemicals, such as flavonoids and carotenoids, which have
antioxidant properties and may help reduce the risk of chronic diseases. Researchers and
nutritionists study the chemical composition and nutritional value of lima beans to better
understand their potential health benefits and to provide evidence-based dietary
recommendations.

1.3 Aim and objectives

The aim of this study is the chemical composition and nutritional value of lima beans. Its
specific objectives include;

2
 i. to determine the proximate and sensory properties of lima beans.
ii. to determine the proximate composition (moisture content, protein, fat,
carbohydrates, fiber, and ash content) of lima beans.
iii. to determine the sensory properties (taste, aroma, texture, and appearance) of lima
beans.
iv. to make comparison between the flour and starch in terms of baking, frying,
cooking etc.

1.4 Significance of the study

The study of the chemical composition and nutritional value of lima beans is important
for several reasons. Firstly, it helps us understand the specific nutrients and compounds
present in lima beans, which can have implications for human health. By knowing the
nutritional value, we can determine if lima beans are a good source of essential
nutrients such as protein, fiber, vitamins, and minerals. Additionally, studying the
chemical composition of lima beans can help us understand their potential benefits and
uses in various industries. For example, if certain compounds in lima beans have
antioxidant or antimicrobial properties, they could be used in the development of new
food products or even pharmaceuticals. Furthermore, understanding the chemical
composition and nutritional value of lima beans can also aid in agricultural practices.
By knowing the specific nutrients required for optimal growth and development of lima
beans, farmers can ensure that they provide the necessary conditions for high-quality
crop production. Overall, studying the chemical composition and nutritional value of
lima beans is crucial for promoting human health, exploring potential industrial
applications, and improving agricultural practices.
1.5 Scope and limitations of study
This research work is limited to the study of the overall quality, nutritional value and
sensory evaluation of lima bean.

3
 CHAPTER TWO
Literature Review
2.1 Origin of lima beans

Phaseolus lunatus is found in Meso- and South America. Two gene pools of cultivated
lima beans point to independent domestication events. The Mesoamerican lima bean is
distributed in neotropical lowlands, while the other is found in the western Andes. [6] They
were discovered in Peru and may have been the first plant that was brought up under
civilization by the native farmers (Leonard & Jonathan, 1970).

The Andes domestication took place around 2000 BC (Molta-Aldana et al., 2010) and
produced a large-seeded variety (lima type), while the second, taking place in
Mesoamerica around 800 AD, produced a small-seeded variety (Sieva type) (Molta-
Aldana et al., 2010). By around 1300, cultivation had spread north of the Rio Grande,
and, in the 1500s, the plant began to be cultivated in the Old World (Molta-Aldana et al.,
2010).

The small-seeded (Sieva) type is found distributed from Mexico to Argentina, generally
below 1,600 m (5,200 ft) above sea level, while the large-seeded wild form (lima type) is
found distributed in the north of Peru, from 320 to 2,030 m (1,050 to 6,660 ft) above sea
level.

The Moche culture (1–800 CE) cultivated lima beans heavily and often depicted them in
their art (Larco & Rafael, 2001). During the Spanish Viceroyalty of Peru, lima beans
were exported to the rest of the Americas and Europe, and since the boxes of such goods
had their place of origin labeled "Lima, Peru", the beans got named as such.[8] Despite the
origin of the name, when referring to the bean, the word "lima" is generally pronounced
differently from the Peruvian capital.

The term "butter bean" is widely used in North and South Carolina for a large, flat and
yellow/white variety of lima bean (P. lunatus var. macrocarpus, or P. limensis. In the
United States Sieva-type beans are traditionally called butter beans, also otherwise known

4
 as the Dixie or Henderson type. In that area, lima beans and butter beans are seen as two
distinct types of beans, although they are the same species. In the United Kingdom and
the United States, "butter beans" refers to either dried beans which can be purchased to
rehydrate, or the canned variety which are ready to use. In culinary use there, lima beans
and butter beans are distinct, the former being small and green, the latter large and
yellow. In areas where both are considered to be lima beans, the green variety may be
labelled as "baby" (and less commonly "junior") limas.

Lima beans (P. lunatus)

2.2 Scientific classification of lima beans (Phaseolus lunatus)

Kingdom: Plantae

Clade: Tracheophytes

Clade: Angiosperms

Clade: Eudicots

Clade: Rosids

Order: Fabales

Family: Fabaceae

Subfamily: Faboideae

Genus: Phaseolus

Species: P. lunatus

2.3 Plant behavior

5
 Phaseolus lunatus has adapted to live in many different climates around the world. One
of these adaptations includes a particularly effective induced herbivory defense. The lima
bean is able to signal to the carnivorous natural enemy of herbivores, the carnivorous
mite, mediated by HIPVs (Herbivore Induced Plant Volatiles) in an attempt to save itself
from further predation (Arimura et al., 2009).

The lima bean plant does this as an induced defense when being eaten by herbivorous
predators. It is the mechanical wounding and chemical elicitors from insect oral
secretions that first begin the signaling pathway to induce HIPV production (Vilvado et
al., 2017). Once this pathway is induced, the plant produces HIPVs which are released in
to the air and can be received by any organisms that have receptors capable of receiving
HIPVs, which includes: carnivores, conspecific and heterospecific herbivores, as well as
neighboring plants. It is this signaling of the carnivorous natural enemy of herbivores that
is of particular interest, as they become attracted to the plant and will then come and prey
upon the plant's herbivorous enemy, thereby reducing herbivory of the plant
(Hettenhausen et al., 2017).

One particular experiment in which this was made apparent was in the understanding of
the tritrophic system between the lima bean plant, two-spotted spider mite, and the
carnivorous mite (Arimura et al., 2009).

Here experimenters noticed an increase in HIPVs when the lima bean plant was preyed
on by the two-spotted spider mite (Arimura et al., 2009). Then, when the carnivorous
mite was introduced, it had increased prey searching efficacy and overall attraction to the
lima bean plant, even once the two-spotted spider mite was removed, but the HIPVs
remained high (Arimura et al., 2009).

2.4 Toxicity

Like many beans, raw lima beans are toxic (containing e.g. phytohaemagglutinin) if not
boiled for at least 10 minutes. However, canned beans can be eaten without having to be
boiled first, as they are pre-cooked (Adeparusi, 2001).

The lima bean can contain anti-nutrients like phytic acids, saponin, oxalate, tannin,
and trypsin inhibitor. These inhibit absorption of nutrients in animals and can cause

6
 damage to some organs. In addition to boiling, methods of roasting, pressure cooking,
soaking, and germination can also reduce the antinutrients significantly (Jayalaxmi et al.,
2015).

2.3 Underutilized legumes

A legume (/ˈlɛɡjuːm, ləˈɡjuːm/) is a plant in the family Fabaceae (or Leguminosae), or

the fruit or seed of such a plant. When used as a dry grain, the seed is also called a pulse.
Legumes are grown agriculturally, primarily for human consumption,
for livestock forage and silage, and as soil-enhancing green manure. Well-known
legumes include beans, soybeans, chickpeas, peanuts, lentils, lupins, grass
peas, mesquite, carob, tamarind, alfalfa, and clover. Legumes produce a botanically
unique type of fruit – a simple dry fruit that develops from a simple carpel and
usually dehisces (opens along a seam) on two sides.

Legumes are notable in that most of them have symbiotic nitrogen-fixing bacteria in
structures called root nodules. For that reason, they play a key role in crop rotation.

Some Fabaceae, such as Scotch broom and other Genisteae, are leguminous but are
usually not called legumes by farmers, who tend to restrict that term to food crops (Apata
et al., 1994).

The term pulse, as used by the United Nations' Food and Agriculture
Organization (FAO), is reserved for legume crops harvested solely for the dry seed
(Ezema, 1989) This excludes green beans and green peas, which are
considered vegetable crops. Also excluded are seeds that are mainly grown for oil
extraction (oilseeds like soybeans and peanuts), and seeds which are used exclusively for
sowing forage (clovers, alfalfa). However, in common usage, these distinctions are not
always clearly made, and many of the varieties used for dried pulses are also used for
green vegetables, with their beans in pods while young.

2.3.1 Uses of lima beans (underutilized legumes)

Cultivated legumes encompass a diverse range of agricultural classifications,

spanning forage, grain, flowering, pharmaceutical/industrial, fallow/green manure, and
timber categories. A notable characteristic of many commercially cultivated legume

7
 species is their versatility, often assuming multiple roles concurrently. The extent of these
roles is contingent upon the stage of maturity at which they are harvested.

[Link] Human consumption

Grain legumes[12] are cultivated for their seeds, which are used for human and animal
consumption or for the production of oils for industrial uses. Grain legumes
include beans, lentils, lupins, peas, and peanuts (Smith & George, 2013).

Legumes are used as a key ingredient in vegan meat and dairy substitutes. They are
growing in use as a plant-based protein source in the world marketplace. [14][15] Products
containing legumes grew by 39% in Europe between 2013 and 2017 (Smith & George,
2013).

[Link] Nutritional value

Legumes are a significant source of protein, dietary fiber, carbohydrates and dietary
minerals; for example, a 100 gram serving of cooked chickpeas contains 18 percent of
the Daily Value (DV) for protein, 30 percent DV for dietary fiber, 43 percent DV
for folate and 52 percent DV for manganese (Onibon et al., 2007).

Legumes are also an excellent source of resistant starch which is broken down
by bacteria in the large intestine to produce short-chain fatty acids (such as butyrate) used
by intestinal cells for food energy (Ikechukwu et al., 2010).

[Link] Forage

Forage legumes are of two broad types. Some, like alfalfa, clover, vetch (Vicia), stylo
(Stylosanthes), or Arachis, are sown in pasture and grazed by livestock. Other forage
legumes such as Leucaena or Albizia are woody shrub or tree species that are either
broken down by livestock or regularly cut by humans to provide livestock feed. Legume-
based feeds improve animal performance compared to a diet of perennial grasses. Factors
to which this is attributed are larger consumption, faster digestion and higher feed
conversion rate (Fasoyiro et al., 2006).

8
 The type of crop(s) grown or animal rearing will be dependent on the farming system,
either vegetables, tubers, grains, cattle etc. In cattle rearing, legume trees such
as Gliricidia sepium can be planted along edges of field to provide shade for cattle, the
leaves and bark are often eaten by cattle. Green manure can also be grown between
periods when crops of economic importance are harvested prior to the next crops to be
planted.

[Link] Other uses

Legume species grown for their flowers include lupins, which are farmed commercially
for their blooms as well as being popular in gardens worldwide. Industrially farmed
legumes include Indigofera and Acacia species, which are cultivated for dye and natural
gum production, respectively. Fallow or green manure legume species are cultivated to be
tilled back into the soil in order to exploit the high levels of captured atmospheric
nitrogen found in the roots of most legumes. Numerous legumes farmed for this purpose
include Leucaena, Cyamopsis, and Sesbania species. Various legume species are farmed
for timber production worldwide, including numerous Acacia species
and Castanospermum australe.

Some legume trees, like the honey locust (Gleditsia) can be used in agroforestry
(Kathirvel & Kumudha, 2011). Others, including the black locust (Robinia
pseudoacacia), Kentucky coffeetree (Gymnocladus dioicus), Laburnum, and the woody
climbing vine Wisteria, have poisonous elements.

2.4 Mung bean (Vigna radiata)

The mung bean (Vigna radiata), alternatively known as the green gram
xanh (Vietnamese; literally, "green bean"), kacang hijau (Indonesian; literally "green
bean") or munggo (Philippines), is a plant species in the legume family (Qayyum et al.,
2012). The mung bean is mainly cultivated in East, Southeast and South Asia.[4] It is used
as an ingredient in both savoury and sweet dishes.

2.4.1 Morphology

Mung bean (Vigna radiata) is a plant species of Fabaceae which is also known as green
gram.[6] It is sometimes confused with black gram (Vigna mungo) for their

9
 similar morphology, though they are two different species (De Oliveira, 2006). The
green gram is an annual vine with yellow flowers and fuzzy brown pods. There are three
subgroups of Vigna radiata, including one cultivated (Vigna radiata subsp. radiata) and
two wild ones (Vigna radiata subsp. Sublobata and Vigna radiata subsp. glabra). It has a
height of about 15–125 cm (AOAC, 2000).

Mung bean has a well-developed root system. The lateral roots are many and slender,
with root nodules grown.[9] Stems are much branched, sometimes twining at the tips.
Young stems are purple or green, and mature stems are grayish yellow or brown. They
can be divided into erect cespitose, semi-trailing and trailing types. Wild types tend to be
prostrate while cultivated types are more erect (Sosulski et al., 1977).

Leaves are ovoid or broad-ovoid, cotyledons die after emergence, and ternate leaves are
produced on two single leaves. The leaves are 6–12 cm long and 5–10 cm
wide. Racemes with yellow flowers are borne in the axils and tips of the leaves, with 10-
25 flowers per pedicel, self-pollinated. The fruits are elongated cylindrical or flat
cylindrical pods, usually 30-50 per plant. The pods are 5–10 cm long and 0.4-0.6 cm
wide and contain 12-14 septum-separated seeds, which are either green, yellow, brown or
blue and can be cylindrical or spherical in shape. [9] Seed colors and presence or absence
of a rough layer are used to distinguish different types of mung bean (Lambrides et al.,
2007).

2.4.2 Scientific classification

Kingdom: Plantae

Clade: Tracheophytes

Clade: Angiosperms

Clade: Eudicots

Clade: Rosids

Order: Fabales

Family: Fabaceae

Subfamily: Faboideae

10
 Genus: Vagna

Species: V. radiate

2.5 Vigna umbellata

Vigna umbellata, previously Phaseolus calcaratus, is a warm-season annual vine legume

with yellow flowers and small edible beans. It is commonly called rice bean. To date, it is
little known, little researched and little exploited (Rahma, 2000). It is regarded as a
minor food and fodder crop and is often grown as intercrop or mixed crop
with maize (Zea mays), sorghum (Sorghum bicolor) or cowpea (V. unguiculata), as well
as a sole crop in the uplands, on a very limited area. Like the other Asiatic Vigna species,
rice bean is a fairly short-lived warm-season annual. Grown mainly as a dried pulse, it is
also important as a fodder, a green manure and a vegetable. Rice bean is most widely
grown as an intercrop, particularly of maize, throughout Indo-China and extending into
southern China, India, Nepal and Bangladesh. In the past it was widely grown as lowland
crop on residual soil water after the harvest of long-season rice, but it has been displaced
to a great extent where shorter duration rice varieties are grown. Rice bean grows well on
a range of soils. It establishes rapidly and has the potential to produce large amounts of
nutritious animal fodder and high quality grain (Ojieh et al., 2008).

2.5.1 Taxonomy

The cultivated Asiatic Vigna species belong to the sub-genus Ceratotropis, a fairly
distinct and homogeneous group, largely restricted to Asia, which has a chromosome
number of 2n = 22 (except V. glabrescens, 2n = 44). There are seven cultivated species
within the sub-genus, including mung bean or green gram (V. radiata), black
gram or urad bean (V. mungo), adzuki bean (V. angularis) and moth bean (V. aconitifolia)
as well as a number of wild species. Artificial crosses have been made between V.
mungo and V. umbellata to produce improved mung bean varieties (e.g. Singh et al.,
2006).

11
 There are three more or less secondary gene pools within the group: ricebean is closer
to V. angularis than to the other species, being in the Angulares group (Kaga et al., 1996,
Tomooka et al., 2003).

Scientific classification

Kingdom: Plantae

Clade: Tracheophytes

Clade: Angiosperms

Clade: Eudicots

Clade: Rosids

Order: Fabales

Family: Fabaceae

Subfamily: Faboideae

Genus: Vagna

Species: V. umbellata

2.5.2 Adaptation and agronomy

Rachie & Roberts (1974), classed ricebean as adapted to subhumid regions with 1000–
1500 mm precipitation, although they noted that other factors were also involved in
adaptation, for example rainfall pattern, moisture distribution, temperature, cloud cover
and relative humidity, soil characteristics, pests and diseases. They noted the importance
of human needs in assessing adaptation – for example taste, the need for a particular use,
or market price. Average yields were between 200 and 300 kg ha−1, although with the
potential for 1200 kg ha−1, the crop would grow on a range of soils, and was resistant to
pests and diseases. It would mature in as little as 60 days, and although performing well
under humid conditions, was also tolerant to drought (NAS 1979) and high temperatures.
It is tolerant to some degree of waterlogging, although the young plants appear to be
susceptible (de Carvalho & Veira, 1996). Ricebean is also known to be tolerant to acid

12
 soils (Dwivedi, 1996). Shattering is a problem in comparison with other grain legumes,
and can be particularly serious under conditions of frequent wetting and drying.

Ricebean is a neglected crop, cultivated on small areas by subsistence farmers in hill

areas of Nepal, northern and northeastern India, and parts of southeast Asia. It can be
grown in diverse conditions and is well known among farmers for its wide adaptation and
production even in marginal lands, drought-prone sloping areas, and flat
rainfed tars (unirrigated, ancient alluvial river fans). It is mainly grown between 700 and
1300 m asl, although in home gardens it is found from 200 up to 2000 m. Most of the
crop currently grown in Nepal is used as food for humans, with a smaller proportion used
for fodder and green manuring.

Generally, ricebean is grown as an intercrop with maize, on rice bunds or on the terrace
risers, as a sole crop on the uplands or as a mixed crop with maize in the khet (bunded
parcels of lands where transplanted rice is grown) land. Under mixed cropping with
maize it is usually broadcast sometime between sowing maize and that crop's first and
second earthing up, so ricebean sowing extends from April–May to June.

Ricebean is valuable for its ability to fix nitrogen in depleted soils and in mixed cropping
with local varieties of maize, as well as for its beneficial role in preventing soil erosion.
The crop receives almost no inputs, and is grown on residual fertility and moisture and in
marginal and exhausted soils. Anecdotal evidence indicates that the area and production
of ricebean in Nepal is declining due to the introduction of high yielding maize varieties
and increasing use of chemical fertilizers, while consumption is decreasing due to
increased availability of more preferred pulses in the local markets. No modern plant
breeding has been done and only landraces with low yield potential are grown. These
have to compete with other summer legumes such as soybeans (Glycine max), black
gram, cowpea, common beans (Phaseolus vulgaris) and horse gram (Mactrotyloma
uniflorum). Other production constraints that limit the production of ricebean include
small and fragmented land holdings and declining productivity.

2.5.3 Morphology

Wild forms are typically fine-stemmed, freely-branching and small-leaved, with a

twining habit, photoperiod sensitivity and indeterminate growth (Lawn, 1995). Flowering

13
 is asynchronous, and there is a tendency to hard seeds. In many areas, landraces which
retain many of these characteristics persist, in particular with regard to daylight
sensitivity, growth habit and hard seeds. Seed color is variable, but commonly red or
yellow.

CHAPTER THREE
Materials and method
3.1 Materials
3.1.1 Equipments
Hammer mill, weighing balance, test tube, retort stand, oven, beaker, conical flask,
vacuum condenser etc.
3.1.2 Reagents
Ethanol, petroleum ether, NaOH, distilled water, methanol
3.2 Method
3.2.1 Collection and Preparation of Samples
The bean samples were sun and oven dried to constant mass, milled to a particle size of
0.2mm using a hammer mill, packaged in a low density polyethylene bag (LDPE) and
kept in a cool dry place pending analysis .
3.2.2 Proximate Analysis
Proximate analysis of the lima bean was carried out to determine crude protein, crude
fibre, total ash, crude fibre, total carbohydrate, crude lipid and moisture content u sing
the methods of AOAC (2000). Triplicate determinations were made whereby the
average and standard deviations are reported.
Determination of Moisture Content

14
2.0 grams of the sample was accurately weighed into a previously cleaned, dried and weighed
crucible. The crucible with its content was put into a Gallenkamp drying oven at 105 oC for 3
hours. The sample was then cooled in desiccators and weighed. The process was repeated until a
constant weight was obtained. The loss in weight expressed as a percentage of the initial weight
of sample gave the percent moisture (AOAC, 2000).

% Moisture = (weight of wet sample – weight of dry sample) x 100

Sample weight

Determination of Ash Content

1 gram of the samples was weighed into a clean dried and cooled crucible. It was incinerated in a
furnace at 550 to 600°C for 3 hours. It was then removed and allowed to cool in desiccators and
weighed again. The percentage ash content was calculated as:

% Ash content = weight of ash (g) x 100

Wt. of original sample

Crude Protein Content Determination

2.0 g of the sample was weighed into a digestion flask containing 0.5g of selenium catalyst. 25
cm3 of concentrated H2SO4 was added and the contents thoroughly mixed. The flask was then
heated on a digestion burner for 8 hours until the solution was green and clear. The solution was
transferred into a 100cm3 volumetric flask and made up to the mark with distilled water. 25cm3
of 2% boric acid was pipette into a 250cm3 conical flask and 2 drops of mixed indicator (20cm3
of bromocresol green and 4cm3 of methyl red) solution were added 10cmZ of the digested
sample solution was then introduced into a Kjeldahl flask, the condenser tip if the distillation
apparatus containing 15cm3 if 40% NaOH was dipped into the boric acid contained in the
conical flask. The ammonia in the sample solution was then distilled into the boric acid until it
became bluish green. The distillate was titrated with 0.1M HCl solution colourless end point.
The percent total nitrogen and crude protein were calculated (AOAC, 2000).

% Total nitrogen = (100 x (VA – VB) x M x 0.01401) x 100

10W

15
Where:

VA = volume (cm3) of HCl used in the sample titration

VB = volume (cm3) of HCl used in the blank titration

M = Molarity of HCl W = weight of sample (g)

% Crude protein = % nitrogen x 6.25

Crude Fibre Content Determination

2.0 g of the defatted sample (from crude fat determination) was transferred into a 250 cm 3
Erlenmeyer flask and 2.5 cm3 of 1.25% H2SO4 was added. The content of the flask was boiled
under reflux and digested for 30 minutes. At the end of the 30minutes, the content was filtered
and subsequently washed with boiling water until the washings were no longer acidic using blue
litmus paper. The sample was washed back into the flask with 200 cm 3 boiling 1.25% NaOH
solution and boiled for 30 minutes. It was then filtered and thoroughly washed with boiling water
until the washings were no longer alkaline using red litmus paper. The crucible with its content
was then dried in an oven at 105 0C overnight and cooled in desiccators and weighed. The
crucible with its content was them ignited in a furnace at 600 0C for 30minutes, cooled and
weighed. The loss in weight was expressed as a percentage of the initial weight of the sample
(AOAC, 2000).

Crude fibre (%) = (Wt. of crucible + sample before ignition) – (weight of crucible + ash) x 100

Weight of fresh sample

Carbohydrate Content Determination

Total percentage carbohydrate (Nitrogen Free Extract) was determined by the difference method
as reported by Amadi et al., (2004). This method involves adding the total values of crude
protein, crude fat, crude fibre, moisture and ash constituents of the sample and subtracting it
from 100. The value obtained is the percentage carbohydrate.

16
% Carbohydrate = 100 – (% moisture + % ash + % protein + % fat + % fibre)

Determination of Crude Lipid Content

5g of the sample was weighed into the extraction thimble, and about 50cm3 of petroleum ether
(40 – 60oC) range was added to the extraction flask. A condenser was fixed at the top of the
extractor. The flask was fitted into the extraction unity and refluxed to about 60oC for 6 hours.
The ether extract was evaporated on an evaporating bath until the lipid was solvent free. This
was dried in an oven at 100oC for 1 hour, cooled in a desiccator and weighed. The lipid was
stored in plastic containers for further analysis.

% Total nitrogen = (weight of beaker + lipid) – (weight of beaker) x 100

Sample weight

3.2.3 Sensory properties

Sensory evaluation was carried out using a 10-man untrained panellist to access the
organoleptic attributes of the prepared samples. The organoleptic attributes assessed
were; taste, aroma, appearance and general acceptability. The panellists were selected
randomly from the students of Taraba state university, Jalingo. The sensitivity evaluation
was conducted using a 9-point hedonic scale as described by (Ihekoronye & Ngoddy,
1985), where the scoring scale range from 9 = liked extremely and to 1 = disliked
extremely.

3.2.4 Comparison between the flour and starch of lima beans in terms of baking, frying
and cooking.

To assess the suitability of the flour and starch for baking, cooking, and frying, various
tests were performed. For baking, you can evaluate the flour and starch's ability to
provide structure, texture, and flavour to baked goods. This can involve conducting tests
such as measuring the volume, crumb texture, and sensory evaluation of baked products
made with the flour and starch.

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 For cooking and frying, the flour and starch's ability is being assess to thicken, bind, and
provide a desirable texture to dishes. This can involve conducting tests such as measuring
the viscosity, gelatinization properties, and sensory evaluation of cooked or fried
products made with the flour and starch.

3.2.4 Statistical analysis

The data were subjected to analysis of variance (ANOVA) test and the differences
between the means were compared for their significance (p<0.05) using SPSS (Statistical
package for social sciences) software v.17 (IBM corporation). The scores given for all the
sensory attributes such as colour, texture, aroma, taste, etc., for each sample were
tabulated. The mean value was calculated for each attribute of a sample, representing the
panel’s judgment about the sensory quality of the product. These were depicted
graphically as sensory profiles.

REFERENCES

Abbey, B.W. and Ibeh G.O. (1998). Functional properties of raw processed cowpea ( vgna
uguiculata) flour. Journal of Food Science 53: 1775-1777.

Adeparusi, E. O. (2001). "Effect of processing on the nutrients and anti-nutrients of lima bean
(Phaseolus lunatus L.) flour". Food/Nahrung. 45 (2): 94–96.

Adeyeye, E.I., Oshodi, A.A. and Ipinmoroti, K.O. (1994), ‘‘Functional properties of some
varieties of African yam bean (Sphenostylis stenocarpa) flour’’, International Journal
of Food Sciences and Nutrition, 45, 115-126.

Allen, O.N and Allen, C.K. (1981). The leguminosae: A source book of characteristics, uses
and modulation. Macmillan Publishers Ltd., London, U.K., pp. 5-29.

AOAC (2000). Association of Official Analytical Chemists. Official Method of Analysis.

13th edition. Washington DC.

Aoki, H., Taneyama, O., Orimo, N. and Kitagawa, I. (1981). Effect of lipophilization of soy
protein on its emulsion stabilizing properties. Journal of Food Science, 46, 1192-1199.

18
Apata, D.F. and Ologhobo, A.D. (1994), “Biochemical evaluation of some Nigerian legume
seeds”, Food Chemistry, 49, 333-338.

Arimura, G.-I.; Matsui, K.; Takabayashi, J. (1 May 2009). "Chemical and Molecular Ecology of
Herbivore-Induced Plant Volatiles: Proximate Factors and Their Ultimate
Functions". Plant and Cell Physiology. 50 (5): 911–923.

Arkroyed W.R. and Doughty J. (1994). Legumes in Human Nutrition. Food and agricultural
Nutrition Studies Publication. Lima Bean (Phaseolus Lunatus). 1:349.

Bampidis, V.A. and Christodoulou, V. (2011) Chickpeas (Cicer arietinum) in animal nutrition:
A review. Animal Feed Science and Technology, 168, 1-20.

Bello-Pe´rez L.A., Sa´yago-Ayerdi S.G., Cha´vez-Murillo C.E., Agama-Acevedo E., Tovar J.,
(2007). Proximal composition and in vitro digestibility of starch in Lima bean
(Phaseolus lunatus) varieties. Journal of food Science and Agriculture, 87, 2570-2575.

Dahl, L.K, (1972) Salt and hypertension, American Journal of Clinical Nutrition, 25, 231-238.

De Oliveira, A. C. (2006). Chemical composition, dietary fibre and resistant starch contents of
raw and cooked pea, common bean, chickpea and lentil legumes. Food Chemistry, 94,
327-330.

Deshpande, U.S., and Deshpande, S.S. (1991), Legumes, In: Foods of Plant Origin:of Plant
Origin: Production, Technology and Human Nutrition (Salunkhe, D.K. and Deshpande,
S.S. eds), New York, USA, Van Nonstrand Reinhold, pp. 137-300.

Dossou, V.M., Agbenorhevi, J.K., Alemawor, F and Oduro, I. (2014) Physicochemical and
Functional Properties of full fat and defatted Ackee (Blighia sapida) Aril
Flours. American Journal of Food Science and Technology, 2(6), pp. 187-191.

Ekpenyong, T.E. and Borchers, R.L. (1980), ‘‘Effect of cooking on chemical composition of
winged bean (Psophocarpus tetragonobolus)’’, Journal of Food Science, 45, 1559-
1560.

19
Ekpo, K.E and Ugbenyen, A.M. (2011). Comparative evaluation of certain functional
properties of four different varieties of Lima Bean (Phaseolus Lunatus) flour, Annals of
Biological Research, 2 (2), 399-402.

Ezema, (1989) Effect of heat on in-vitro digestibility of navy beans ([Link]). Michigan
Quaterly Bull 46: 87.

Fasoyiro. S.R., Ajibade, A.J. Omole, O.N. Adeniyan and Farinde, E.O. (2006). Proximate,
minerals and antinutritional factors of some underutilized grain legumes in south-
western Nigeria, Journal Of Institute Of Agricultural Research And Training, Obafemi
Awolowo University, P.M.B 5029, Moor-Plantation, Ibadan, Nigeria Nutrition and
Food Science, 36(1),18-23.

Granito M., Brito Y., Torres A., (2007). Chemical composition, antioxidant capacity and
functionality of raw and processed (Phaseolus lunatus). Journal of food Science and
Agriculture, 87, 2801-2809.

Hettenhausen, Christian; Li, Juan; Zhuang, Huifu; Sun, Huanhuan; Xu, Yuxing; Qi, Jinfeng;
Zhang, Jingxiong; Lei, Yunting; Qin, Yan; Sun, Guiling; Wang, Lei; Baldwin, Ian T.;
Wu, Jianqiang (8 August 2017). "Stem parasitic plant Cuscuta australis (dodder) transfers
herbivory-induced signals among plants". Proceedings of the National Academy of
Sciences. 114 (32): E6703–E6709.

Iheanacho, K.M.E., (2010). Comparative Studies of the Nutritional Composition of Soy Bean
(Glycine Max) and Lima Bean (Phaseolus Lunatus). Scientia Africana, 9(2), 29-35.

Ikechukwu Edwin Ezeagu, Madu Daniel Ibegbu., (2010). Biochemical Composition and
Nutritional Potential of Ukpa: A Variety of Tropical Lima Beans (Phaseolus Lunatus)
From Nigeria – A Short Report Pol. Journal of Food Nutrition and Science, 60, 231-
235.

Jayalaxmi, Baddi; Vijayalakshmi, D.; Usha, Ravindra; Revanna, M. L.; Chandru, R.; Gowda, P.
H. Ramanjini (21 December 2015). "Effect of different processing methods on
proximate, mineral and antinutrient content of lima bean (Phaseolus lunatus) seeds”.

20
Kathirvel P and Kumudha P, (2011). A Comparative Study on the Chemical Composition of
Wild and Cultivated Germplasm of Phaseolus Lunatus L. International Journal of
Applied Biology and Pharmaceutical Technology, 2(4), 298.

Kaur M, and Singh N (2006). Relationships between selected properties of seeds, flours, and
starches from different chickpea cultivars. International Journal of Food
Properties, 9:597-608.

Kaur, S., Singh, N., Sodhi, N. S., and Rana, J. C. (2009). Diversity in properties of seed and
flour of kidney bean germplasm. Food Chemistry, 117, 282-289.

Lambrides, C. J.; Godwin, I. D. (2007), Kole, Chittaranjan (ed.), "Mungbean", Pulses, Sugar and
Tuber Crops, Genome Mapping and Molecular Breeding in Plants, Berlin, Heidelberg:
Springer, vol. 3, pp. 69–90.

Larco Hoyle, Rafael. Los Mochicas. Museo Arqueológico Rafael Larco Herrera. Lima 2001.

Leonard, Jonathan Norton (1970). Recipes, Latin American cooking. Time-Life International
(Nederlands). p. 21.

Luis Chel-Guerrero, Mario Domínguez-Magaña, Alma Martínez-Ayala, Gloria Dávila-Ortiz,

and David Betancur-Ancona, (2012). Lima Bean (Phaseolus lunatus) Protein
Hydrolysates with ACE-I Inhibitory Activity. Food and Nutrition Sciences, 3, 511-521.

Mackie, W.W. (1983). Origin, dispersal and variability of the Lima bean Phaseolus
[Link], 15: 1-29.

Milán-Carrillo, J., Reyes-Moreno, C., Armienta-Rodelo, E., CaráH Bez-Trejo, A. and Mora-
Escobedo, R. (2000). Physicochemical and nutritional characteristics of extruded flours
from fresh and hardened chickpeas (Cicer arietinum L). Lebensmittel-Wissenschaft
und-Technologie, 33, 117-123.

Moses O, Olawuni I. and Iwouno J.O. (2012). The Proximate Composition and Functional
Properties of Full-Fat Flour, and Protein Isolate of Lima Bean (Phaseolus
Lunatus). 1:349.

21
Motta-Aldana, Jenny R.; Serrano-Serrano, Martha L.; Hernández-Torres, Jorge; Castillo-
Villamizar, Genis; Debouck, Daniel G.; Chacóns, Maria I. (September 2010). "Multiple Origins
of Lima Bean Landraces in the Americas: Evidence from Chloroplast and Nuclear DNA
Polymorphisms". Crop Science. 50 (5): 1773–1787.

Naczk, M., Diosady, L. L., & Rubin, L. J., (1985). Functional properties of canola meals
produced by a two-phase solvent extraction system. Journal of Food Science, 50, 1685-
1692.

Oboh, H.A and Omofoma, C.O. (2008). The effects of heat treated lima bean ( Phaseolus
lunatus) on plasma lipids in hypercholesterolaemic rats. Pakistan Journal of
Nutrition, 7(5), 636-639.

Ojieh G.C., Oluba O.M., Ogunlowo Y.R., Adebisi K.E., Eidangbe G.O. and Orole R.T.,
(2008). Compositional studies of Citrullus lanatus (Egusii melon) Seed. The Internet
Jouurnal of Nutrition and Wellness, 6(1), 511-521.

Omueti, O. and Singh, B.B. (1987) ‘‘Nutritional attributes of improved varieties of cowpea
(Vigna unguiculata L. Walp)’’, Food Science and Nutrition, 41,103-112, 1559-1560.

Qayyum, M. M. N., Butt, M. S., Anjum, F. M. and Nawaz, H. (2012) Composition analysis of
some selected legumes for protein isolates recovery, The Journal of Animal & Plant
Sciences, 22(4), 1156-1162.

Rahma E.A, (2000) Production and Evaluation of Yambean (Hochstex Rich) And Bambara
Groundnut (Voandzeia Subterranean L) Journal of Science and Food Agriculture, 41,
123-124.

Rickard, J.E., Blanshard, J.M.V., and Asaoka, M. (1992). Effects of cultivar and growth season
on the gelatinization properties of cassava (Manihot esculenta) starch, Journal Food
science and Agriculture. (59) 53-58.

Sathe, S. K., Deshpande, S. S., and Salunkhe, D. K. (1982) Functional properties of winged
bean (Psophocarpus tetragonolobus, L.) proteins. Journal of Food Science, 47, 503-
508.

22
Sathe, S.K. and Salunkhe, D.K (2006). Functional properties of the green northern bean
(phaseolus vulgari L) proteins: emulsion, foaming, viscosity and gelation
properties. Journal of Food Science, 46, 71-81.

Smith, George (2018-02-13). "Pulse raising: Grain legume market grows by 37 per cent in
Europe - Meat substitutes have helped drive soaring market interest in pulses, or grain
legumes, across Europe, a recent study has found". New Food Magazine. Retrieved 2021-
03-08.

Sosulski, F.W. and Fleming, S.E., (1977). Chemical, functional, and nutritional properties of
sunflower protein products. Journal of the American Oil Chemists’ Society, 54, 100A.

Sreerama, Y. N., Sashikala, V. B., Pratape, V. M., and Singh, V. (2012). Nutrients and
antinutrients in cowpea and horse gram flours in comparison to chickpea flour:
evaluation of their flour functionality. Food Chemistry, 131, 462-468.

Tharanathan, R. N., and Mahadevamma, S. (2003). Grain legumes, a boon to human nutrition.
Trends in Food and Science Technology, 14, 507-551

Vivaldo, Gianna; Masi, Elisa; Taiti, Cosimo; Caldarelli, Guido; Mancuso, Stefano (8 September
2017). "The network of plants volatile organic compounds". Scientific Reports. 7 (1):
11050.

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