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Discoveries in Agriculture and Food Sciences - Vol. 12, No. 2

Publication Date: April 25, 2024

DOI:10.14738/dafs.122.16777.

Céspedes, M. A. L., & Ascheri, J. L. R. (2024). Prediction, by Mathematical Model, of the Effects of Hydration for Germination at

Different Times and Temperatures on the Nutritional and Functional Characteristics of Chickpea Flour (Cicer Arietinum L.). Discoveries

in Agriculture and Food Sciences, 12(2). 01-26.

Services for Science and Education – United Kingdom

Prediction, by Mathematical Model, of the Effects of Hydration for

Germination at Different Times and Temperatures on the

Nutritional and Functional Characteristics of Chickpea Flour

(Cicer Arietinum L.)

Miguel Angel Larrea Céspedes

Facultad de Ciencias Agropecuarias - Escuela de Ingeniería en Industrias

Alimentarias Universidad Nacional Jorge Basadre Grohmann, Perú

José Luis Ramírez Ascheri

Embrapa Food Technology, and Federal Rural of Rio de Janeiro, (PPGCTA), Rio de

Janeiro, RJ, Brazil

ABSTRACT

The objective of this work was to study the effects of germination time and

temperature on chickpeas by determining appropriate mathematical models

corresponding to each dependent variable to predict the behaviour of the process

and improve the physical, chemical, and functional properties of the flour obtained.

Chickpea grains were subjected to germination processes at the laboratory, after a

hydration process, to study the physical, chemical, and functional characteristics of

the flour obtained from the legume. The Response Surface methodology was used

by the central composite rotational layout. For the soaking process of the grains, the

independent variables were considered, with their respective levels: time (6, 8, 12,

16, and 18 hours) and temperature (18, 20, 25, 30, and 32 °C), being evaluated

through percent moisture as a dependent variable. During the germination process,

the independent variables time (14, 24, 48, 72, and 82 hours) and germination

temperatures (23, 25, 30, 35, and 37 °C) were controlled, keeping the relative

humidity fixed (95%). All mathematical models determined were suitable for

calculating values for each of the variables studied. The best response (R)

operational R1, total protein content (TPC); R2, total lipid content (TLC); R3, total

sugar content (TSC); R4, reducing sugar content (RSC) and R5, total ash content

(TAC). Likewise, the following functional properties were evaluated: R6, water

absorption index (WAI); R7, lipid absorption index (LAI); R8, water solubility index

(WSI); R9, apparent density (APD); and finally, the presence of antinutrients was

studied through R10, urease activity (UA).

Keywords: functional properties, germination effects, proximal composition, urease total,

reducing sugar.

INTRODUCTION

Chickpea is a legume with a high content of proteins and fibres, and its consumption

recommended due to the number of benefits it provides as a bioactive component due to the

diversity of available derived products and their applications in food systems [1]. So much

success is not for nothing. The legume – from the same family as lentils, peas, soybeans, and

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Discoveries in Agriculture and Food Sciences (DAFS) Vol 12, Issue 2, April- 2024

Services for Science and Education – United Kingdom

beans – is highly nutritious and versatile, serving as the basis for several recipes. The regular

consumption is relational to several health benefits: it helps control blood sugar levels, prevent

heart disease, boosts digestion, and even increases satiety – a good ally for those who want to

lose weight, as pointed out in a review of studies published in the American Journal of Clinical

Nutrition [2]. The survey compared 21 studies and found that volunteers who consumed at

least one daily portion (approximately 130g) of legumes as soon as chickpeas showed a

reduction in the scale after six weeks – even without dieting! It has high protein content. It is an

adequate option for those who want to reduce their consumption of protein of animal origin

(such as red meat, poultry, and fish) as it provides a good supply of the nutrient – around 9 g

for every 100 g of cooked grain. Food improves mood, considered by some as the “grain of

happiness” it contains tryptophan, fundamental amino acid in the production of serotonin (a

neurotransmitter linked to the feeling of well-being). The magnesium in legumes is a nutrient

that can help relax and reduce anxiety.

The cereal is packed with iron, a mineral that helps prevent anemia and can help alleviate

symptoms like fatigue and headaches. In more severe cases, these symptoms can affect

cognitive function. Furthermore, it also supports brain health by effectively neutralizing the

harmful effects of free radicals with its high levels of antioxidants. With this, it acts to protect

the brain and stimulates its proper functioning. Legumes reduce the risk of type 2 diabetes;

their low glycemic index (GI) helps control blood glucose levels and lower insulin release.

Therefore, research such as that published in The American Journal of Clinical Nutrition

associates the food with a reduced risk of type 2 diabetes. Its fibres are valuable. Chickpeas

(especially if they are in their shells) are absorbed more slowly in the digestive system, leaving

the consumer feeling fuller for longer. Fibres also improve bowel function. Strengthens the

immune system like other legumes, grains are rich in zinc and vitamins A and E, fundamentals

agents in combating flu and colds, ensuring reinforcement of the immune system. It maintains

healthy bones and teeth; chickpeas are rich in calcium and contain generous amounts of vitamin

K and phosphorus. Together, the nutrients help to strengthen bones, which can prevent

fractures and diseases that affect the bones, such as osteopenia and osteoporosis. The grain acts

in the prevention of cardiovascular diseases, helping to reduce triglycerides and bad

cholesterol related to the obstruction of blood vessels. Furthermore, the potassium in food has

a vasodilating effect, which helps to reduce blood pressure and protect heart health [3], [4], [5],

[6]. India is the largest chickpea producer in the world with 11.380.000 tonnes production per

year. Australia comes second with 998,231 tonnes of yearly production. With 630.000 tonnes

of production per year, Turkey is the third largest producer of chickpeas [7]. Comparatively, in

countries like India, the Middle East, and South American countries, chickpea consumption is

not very significant. To solve the problems related to post-harvest losses of this legume, and to

promote better use, numerous investigations have been carried out. Thus, for example, studies

carried out showed that it is possible to develop a DIP-type product (Diversified Innovative

Products), made from chickpeas and dehydrated tomatoes to promote the consumption of

healthy foods, rich in vitamins, proteins, minerals, and other health benefits [8]. According to

[9], the American Association of Clinical Chemistry (AACC), in 2008, defined sprouted grains as

“Malted or sprouted grains containing all bran, germ and endosperm. They originate and are

considered whole grains. Since the growth of the germination does not exceed the compression

of the grain and the nutritional values are not lower in comparison with germinated”. To

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Céspedes, M. A. L., & Ascheri, J. L. R. (2024). Prediction, by Mathematical Model, of the Effects of Hydration for Germination at Different Times and

Temperatures on the Nutritional and Functional Characteristics of Chickpea Flour (Cicer Arietinum L.). Discoveries in Agriculture and Food Sciences,

12(2). 01-26.

URL: http://dx.doi.org/10.14738/dafs.122.16777

germinate chickpeas, first wash them, soak them with double the volume of water, for 8 to 12

hours, place a damp cloth at the base of a tray, spread them out, and cover them with another

damp cloth. Approximately every 8 hours, spray both the chickpeas and the cloth with water,

remove them so that the humidity is uniform, and cover again [10]. The grains should be kept

moist but without excess water. Repeat this operation until the 1 to 2 cm root emerges. For a

seed to germinate, certain conditions must be favourable, such as an adequate amount of water,

and a desirable temperature, from 10 °C the chickpea is capable of germinating, although the

optimal germination temperature ranges between, 25 to 35 °C. The objective of this work was

to study the effects of germination time and temperature of chickpeas by determining

appropriate mathematical models corresponding to each dependent variable, to predict the

behaviour of the process to improve the physical, chemical, and chemical properties

functionalities of the flour obtained.

MATERIAL AND METHODS

Raw Material

Chickpea bean grains were used, produced in the city of Camaná, Peruvian origin, with good

characteristics for human consumption, obtained from the Grau Wholesale Market of Tacna -

Peru located in the enclosure of the Tacna city.

Experimental Details

Figure 1 shows the main stages and controls that were carried out during the development of

the experiment.

Figure 1: Main stages and controls that were carried out during the development of the

experiment.

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Discoveries in Agriculture and Food Sciences (DAFS) Vol 12, Issue 2, April- 2024

Services for Science and Education – United Kingdom

Production of Germinated Chickpea Flour in the Laboratory

Pre-treatment: Immersion of chickpea grains in 1.5% (v/v) sodium hypochlorite solution for

10 minutes to prevent the proliferation of microorganisms, mainly fungi. The grains were then

washed with plenty of water to remove sodium hypochlorite residues, as suggested by [11].

Soaking or hydration: It was carried out with distilled water, for this purpose, 250 g of

chickpeas were placed in a glass container with a capacity of one litter. The best hydration

conditions were studied according to the experimental design described in Table 1. The

variables and levels of variation were temperature and time, keeping the raw material/water

ratio constant (1:3 w/v) until the grains were completely saturated.

Experimental Outline for the Soaking Process and Germination Process

Table 1 shows the variables that were studied, with the respective levels of variation. Planning

22 with a central point was used [12]; the variables were established at three levels coded as -

1, 0, + 1. The actual values were calculated according to the following equation: xi = (Xi - Y)/ΔXi;

Where: xi = encoded value of the variable Xi; Xi = real value of the variable; Y = real value of the

variable at the centre point; ΔXi = differential between two consecutive values. This type of

planning requires two levels of axial variables coded as - and +. The value of  is a function

of the number of independent variables (k = 2, for the present case), which is defined by the

following equation:

 = (22) = x2 + x2 = 1.4142

The present statistical design requires 11 treatments, of which four are factorial (combinations

between levels –1 and +1), two axial (one variable at level  and two at zero) and three central

(at level zero). Which serve to estimate the experimental error and determine the precision of

the polynomial equation [13]. Table 1 presents the composite central rotational experimental

outline for two variables and five levels used in the soaking or hydration process of chickpea

grains, and Table 2 show the effects of temperature and germination time were studied using a

composite rotational central planning (star planning) of the [14] methodology.

Table 1: Levels of the independent variables to be studied during the soaking or

hydration process of chickpea grains.

Variables Variable levels

Independent - = -1.4142 -1 0 1 +  = +1.4142

X1 18 20 25 30 32

X2 6 8 12 16 18

Note. X1, soaking temperature (° C) and X2, soaking time (hours).

Table 2: Levels of the independent variables studied during the germination process of

chickpea grains (star planning).

Variables Variable levels

Independent - = -1.4142 -1 0 1 +  = +1.4142

X1 23 25 30 35 37

X2 14 24 48 72 82

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Céspedes, M. A. L., & Ascheri, J. L. R. (2024). Prediction, by Mathematical Model, of the Effects of Hydration for Germination at Different Times and

Temperatures on the Nutritional and Functional Characteristics of Chickpea Flour (Cicer Arietinum L.). Discoveries in Agriculture and Food Sciences,

12(2). 01-26.

URL: http://dx.doi.org/10.14738/dafs.122.16777

Note, X1, germination temperature (o C), and X2, germination time, (h).

For each experiment, the dependent variables indicated below were evaluated (R, response):

R1 = Total protein content (TPC); R2 = Total lipid content (TLC); R3 = Total sugar content (TSC);

R4 = Reducing sugar content (RSC); R5 = Total ash content (TAC). R6 = Water absorption index

(WAI); R6 = Lipid absorption index (LAI). R7 = Water solubility index (WSI). R8 = Apparent

density (AD) and R9 = Urease activity (UA).

Drying and milling: For drying, it was carried out in an oven at atmospheric pressure at 55 ° C

for a period of 24 hours, until reaching an approximate 12% moisture. Milling: For experimental

purposes and for comprehensive use, the unsprouted and sprouted chickpea grains were

ground in a disc mill without removing the outer covering and the roots that came from

germination. The samples were then subjected to a size reduction operation with an OSTER

blender, at speeds: medium and maximum; they were then sieved using a 500-μm mesh. Finally

placed in glass jars and closed hermetically.

Data Processing and Analysis

For the analysis of the combined effects of the independent variables (X1) and (X2) in the

evaluated responses (Yi), the response surface methodology was used [14]. The averages of the

data from each of the 11 experiments for each response were treated by multiple region

analysis, to obtain first or second order mathematical models containing the linear, quadratic

and product terms of the interactions. In the present case, significant effects were treated by

ANOVA. In it, the degree of significance of the region and the lack of adjustment was observed,

at 95% confidence, with the help of the F test, and the analysis of the coefficient of

determination (R2). The model, to be considered predictive to describe a certain characteristic

in the analysed region, must present significant regression at the 95% confidence level, non- significant lack of adjustment at the same confidence level and a high R2 value (very close to 1

or 100%). In order to identify the experimental area that would allow maximizing some

properties studied, and minimizing others, according to the tendency of each of them, a new

dependent variable was established, based on the models developed by [15]. The mathematical

expression of this multi-response function (MRF) was:

MRF = Superior response product/ Inferior response product

Proximal Composition

Moisture was determined according to method 44-15 of the American Association of Cereal

Chemists [16]; protein, by the Kjeldahl micro distiller method 46-13 of the AACC (1990) using

the factor 6.25; Lipids, according to the Soxhlet method (method 30 –10) AACC [16]; ashes,

method 08–01 established by the AACC [16].

Total Sugars

The Sulfuric Phenol method, according to recommendations established by Dubois [17] in

which glucose is used as a standard solution, determining the absorbance at 490 nm, carried

them out.