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Amino Acid Profile and Physicochemical Properties of African Locust Bean

(Parkia biglobosa) Seeds as affected by Combined Irradiation and Cooking

Article in International Journal of Environment Agriculture and Biotechnology · February 2024

DOI: 10.22161/ijeab.91.16

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 International Journal of Environment, Agriculture and Biotechnology
Vol-9, Issue-1; Jan-Feb, 2024
Peer-Reviewed International Journal
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Journal DOI: 10.22161/ijeab

Amino Acid Profile and Physicochemical Properties of

African Locust Bean (Parkia biglobosa) Seeds as affected
by Combined Irradiation and Cooking
Alex O Asunni1, Stephen A. Fagbemi2, Ajibola M. Oyinloye3, Catherine B. Gowon4 and
Victor N. Enujiugha4*
1Department of Applied Disaster and Emergency Studies, University of Brandon, Brandon MB, R7A6A9 Canada
2NationalAgency for Food and Drug Administration and Control, Lagos, Nigeria
3Department of Food Science and Technology, Federal University, Oye-Ekiti, Nigeria
4Department of Food Science and Technology, Federal University of Technology, Akure, Nigeria.

*Correspondence: [email protected]

Received: 20 Dec 2023; Received in revised form: 05 Feb 2024; Accepted: 12 Feb 2024; Available online: 20 Feb 2024
©2024 The Author(s). Published by Infogain Publication. This is an open access article under the CC BY license
(https://creativecommons.org/licenses/by/4.0/).

Abstract— This study examined the impact of cooking, Co-60 gamma irradiation (5 kGy and 10 kGy) and
combined irradiation (10 kGy) and cooking treatments on the amino acid profile and physicochemical
properties of African locust bean (Parkia biglobosa) seeds, as well as the quality of the seed oil. The treated
and untreated (control) seeds were mechanically dehulled and milled into flour using an attrition mill, and
subsequently analysed for chemical composition, amino acid profile, functional properties and quality of the
seed oil. The results showed a significant increase in the fat content (p ≤ 0.05) after cooking and post-
irradiation cooking treatments. The cooked seed samples recorded a 3.84% increase in total mineral content
although there was a loss in the total percentage minerals after combined irradiation and cooking which
could be linked to leaching of soluble minerals into cooking water. The 10 kGy irradiation dose improved
the total essential amino acids present in the seed possibly due to the lyses of adherrent microflora.
Percentage cysteine was highest in the 5 kGy irradiated seed sample with about 4.8% increase compared to
control. There was a 92.6% loss in foaming capacity of the seed flour as a result of processing; however, 5
kGy irradiated sample recorded only about 26% loss in the foaming capacity. The anti-nutrients were
significantly reduced by cooking, irradiation, post-irradiation cooking up to about 60%, 40%, 26.7% and
60% for cooked, 5 kGy irradiated, 10 kGy irradiated and post-irradiated cooked samples, respectively.
Tannin content was not significantly affected (p<0.05) by the irradiation treatments; it was however reduced
significantly by post-irradiation cooking. The 5 kGy and 10 kGy doses did not significantly affect iodine
value of the oil sample. There was however an increase in iodine value as a result of cooking and post-
irradiation cooking with values as high as 163.53 g I2 and 182.77 g I2 per 100g, respectively. Overall, it
could be concluded that medium dose gamma irradiation did not negatively affect the quality of the African
locust bean seed, which is desirable for elongation of storage life of the seed.
Keywords— Gamma irradiation, Cooking, Locust bean seed, Physicochemical properties, Oil quality.

I. INTRODUCTION led to wider exploration and upgrading of traditional foods

The quest for greater value-addition, improved aesthetics which hitherto were consumed at the basic subsistence level
and elongation of shelf-life of unconventional and wild (Enujiugha, 2000; Oguntimehin et al., 2023). Different
uncultivated forest seeds with high nutrient potentials has techniques are employed to preserve such seeds, one of
which is food irradiation that involves exposure of food to

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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

a controlled source of ionizing radiation with a view to exploration of plant protein and vitamin substitutes, the
reducing microbial load, extending product shelf life, and African locust bean (Parkia biglobosa) has become very
disinfesting agricultural produce (Enujiugha, et al., 2012; popular especially in the fermented ‘Dawadawa’ and ‘iru’
2023a; Oyinloye et al., 2023). Food irradiation is a more forms, which are commonly-derived products from the
reliable and safer technique of preserving food and seeds (Enujiugha, 2009; Gernah et al., 2007). Dawadawa or
improving the nutritional value (Diehl, 2002; Al-Kaisey et iru (fermented Parkia biglobosa seeds) is used for
al., 2002). Three types of radiation are applicable for use in seasoning of traditional soups in all parts of Nigeria and
food preservation: gamma rays, X-rays and rays from high indeed the West Coast of Africa (Enujiugha et al., 2006).
energy electron beams (James, 2006). Gamma irradiation, The sticky, sour solid contains sufficient amounts of
also known as cold pasteurization, is a technique of food protein, vitamins, energy (Oyedokun et al., 2016; 2020) and
preservation that has been proven to prevent insect has appreciable shelf life even without refrigeration.
infestation in food products during storage and also deter Combined irradiation and cooking treatments could further
food contamination by microorganisms (Farkas, 1990). extend the shelf life but have also been shown to effect
Combining low-dose gamma irradiation with cooking in various changes in the physical, chemical and functional
food processing and preservation is a hurdle strategy that properties of foods (Olotu et al., 2014a). This study
allows effective processing while minimizing the severity examined the effect of combined low-dose gamma
of treatment (Enujiugha et al., 2012). To maximize the irradiation and post-irradiation cooking on the functional
effectiveness of combined treatment, mild irradiation and physicochemical properties of African locust bean seed
between 1 and 10 kGy is proposed as best (Campbell-Platt and its oil quality characteristics, in comparison to single
and Grandson, 1990). The use of cooking and γ-irradiation irradiation and cooking treatments. This was with a view to
as forms of preservation are both proven technologies and highlighting the potentials of such hurdle treatments in the
the effectiveness of both could be maximized if they are preservation of the nutrient potentials of the seeds while at
combined in a hurdle effect (Olotu et al., 2014a,b). the same time achieving the twin functions of disinfestation
The main drive in the application of hurdle technology, and and processing.
in this case, combination of low dose gamma irradiation
with cooking, has been the prevalent food safety concerns II. MATERIALS AND METHODS
and low shelf life of our indigenous foods. Fagbemi et al.
Material Procurement and Preparation
(2023) observed the high microbial density in selected
indigenous foods available for direct human consumption in Wholesome African locust bean seeds (Parkia biglobosa)
a typical urban neighbourhood in sub-Saharan Africa, were obtained from a local market (Oja Oba), in Akure,
which is a public health concern when the huge numbers of Ondo State of Nigeria. Upon reception, the seeds were
consuming populations are considered. It is widely believed visually inspected and defective ones were discarded. The
that irradiation of agricultural produce at low doses, before seeds were then kept in air tight polyethylene containers in
storage and processing, could reduce the normally-high a dry and cool environment until ready for use. All the
incidence of microbial infestation and curtail the occurrence chemicals and reagents used in the study were of analytical
of disease-causing pathogenic micro-organisms. grade. The raw grains were treated at room temperature with
Combination of this preservation technique with other gamma rays at doses of 5 and 10 kGy using a multipurpose
treatments like cooking in a hurdle effect would ensure gamma irradiator with a cobalt 60 source (compact-type
effectiveness without adversely affecting the innate commercial radiator) at the Shedan Science and Technology
nutrients. Complex (SHESTCO), Abuja Nigeria (Oyinloye et al.,
2023). Raw untreated locust bean seeds served as control in
Low income earners in developing countries like Nigeria
the experiment. The 5 kGy sample was milled immediately
can hardly afford animal products, such as milk, meat, fish
while the 10 kGy irradiated sample was divided into two
and eggs, which are rich sources of protein. In these areas,
parts; a part was milled while the other part and half part of
staple diets consist mainly of cereal grains or starchy root
the raw (uncooked and un-irradiated) samples were
and tuber crops thus leading to various health problems
subjected to hydrothermal treatment in the proportion of 1:3
associated with protein and vitamin/mineral deficiencies
(seed to water ratio) for 6 h at 100 oC. The samples were
(Enujiugha, 2020). This has led to various researches being
placed in aluminum trays and dried with forced circulation
directed towards the utilization of plant protein sources in
of air at 50-55 oC until constant weight (approximately 24
the form of nuts, oilseeds and legumes to tackle the
h).
prevailing protein-energy malnutrition ravaging many parts
of less-developed nations (Enujiugha and Ayodele-Oni,
2003; Enujiugha et al., 2023b; Talabi et al., 2023). In the

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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

Determination of Proximate chemical composition was dissolved with 5 ml of acetate buffer (pH 2.0) and
Quantitative composition was determined on each of the stored in plastic specimen bottles, which were kept in the
flour samples using the following analytical methods: freezer. The amount of hydrolysate loaded into the TSM
Moisture content using the air oven method at 105 oC until analyser was between 5 to 10 microliters. This was
constant weight was achieved (AOAC, 2012); crude protein dispensed into the cartridge of the analyzer. The TSM
using the micro-Kjeldahl apparatus (AOAC, 2012); crude analyser is designed to separate and analyze free acidic,
fat extracted overnight in a Soxhlet extractor with n-hexane neutral and basic amino acid of the hydrolysate. The period
and quantified gravimetrically; crude ash via exhaustive of an analysis lasted for 76 minutes. All determinations
combustion in a Muffle furnace at 550 oC for 8 h (AOAC, were carried out in triplicates.
2012); crude fibre estimated after digesting known weights Determination of functional properties
of fat-free samples in refluxing 1.25% sulphuric acid and The determination of water and oil absorption capacities
1.25% sodium hydroxide; and carbohydrate determined by followed a modification of the method of Prinyawiwatkul et
difference method (subtracting the percent crude protein, al. (1999). Each flour sample (5 g) was thoroughly mixed,
crude fibre, crude fat, and ash from 100% dry matter). All without pH adjustment with 25 ml of deionized water or oil
analyses were carried out in triplicates. The gross energy in 50 ml centrifuge tubes. Suspensions were stirred
was calculated based on the formula reported by Enujiugha intermittently over a 30 min period at room temperature
and Ayodele-Oni (2003). (28±2 oC) and then centrifuged at 12,000 x g for 30 min at
Mineral analysis 25 oC. The volume of decanted supernatant was centrifuged
Analysis of sodium and potassium contents of the samples and the water and oil absorption capacities were calculated.
was carried out using flame photometer, while phosphorus Triplicate samples were analyzed for each flour sample
was determined colourimetrically by the phosphovanado- category.
molybdate (yellow) method (AOAC, 2012). The other Least gelation concentration was carried out as described by
elemental concentrations were determined after wet Enujiugha and Akanbi (2005). Triplicate suspension of 1-
digestion of sample ash with a mixture of nitric and 20% seed flour sample (dry w/v at 1% increment) were
perchloric acids (1:1 v/v), using atomic absorption prepared in 10 ml of deionized water and mixed thoroughly
Spectrophotometer (AAS, Buck Model 20A, Buck without pH adjustment. The slurries were heated in 125 x
Scientific, East Norwalk, CT06855, USA). All the 20 mm screw-capped test tubes in a water bath with in-built
determinations were carried out in triplicates. magnetic stirrer (Julabo Model SW22, Julabo Labortechnik
Determination of Amino Acids Profile GMBH, Seelbatch, Germany) at 95 ± 2 oC. After 1 h of
heating, tubes were immediately cooled in tap water for 30
The amino acids profile in the sample was determined using
s and then in ice water for 5mins to accelerate gel formation.
the method of Olotu et al. (2014b), with slight
All tubes were then held at 4 oC for 3 h. The least gelation
modifications. The samples were dried to constant weight,
concentration (%) was determined as the concentration
exhaustively defatted, acid-hydrolyzed (or alkaline
above which the sample remained in the bottom of the
hydrolyzed, in the case of tryptophan), evaporated in a
inverted tube.
rotatory evaporator and loaded into the technicon sequential
multi sample amino acid analyzer (TSM). Briefly, a known The foaming properties of the samples were determined
weight of sample powder was weighed into extraction using the method of Coffmann and Garcia (1977), with
thimble and any remaining fat was extracted with slight modifications. Two grams (2 g) of the sample was
chloroform/methanol (2:1) using Soxhlet extraction weighed into 60 ml distilled water in a 100 ml cylinder.
apparatus as described by AOAC (2012); the extraction Solid material was dispersed with spatula and the
lasted for 15 hours. A known weight of the defatted sample suspension was whipped for 5 min using ultra-Turax T25
was then weighed into glass ampoule. Exactly 7 ml of 6N mixer at high speed. Volumes before and after whipping
HCL (or 6N KOH) was added and oxygen was expelled by were noted and volume increase due to whipping was
passing nitrogen into the ampoule in order to avoid possible calculated. The volume of foam in the standing cylinder was
oxidation of some amino acids during hydrolysis e.g. also recorded for foam stability at 1, 5, 10, 20, 30, 60, 90,
methionine and cysteine. The glass ampoule was then 120 and 180 min after whipping. The results were expressed
sealed with Bunsen burner flame and put in an oven preset in percentages (g/g basis).
at 105 ± 5 oC for 22 hours. The ampoule was allowed to cool Emulsifying properties were determined with a slight
before broken open at the tip arid the content was filtered to modification of the method described by Ige et al., (1984).
remove the remains. The filtrate was evaporated to dryness A known quantity (1.8 g) of sample was dispersed in 25 ml
at 40 oC under vacuum in a rotator evaporator. The residue distilled water and 25ml vegetable oil (pure groundnut oil)

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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

was added. The 50 ml mixture was emulsified at high speed Determination of seed oil characteristics
using ultra-Turax T25 mixer for 1 min. emulsion was filled The seed oils of the samples were extracted using Soxhlet
into centrifuge tubes and centrifuged for 5 min at 1,300 x 6 apparatus (Talabi and Enujiugha, 2014) and the rancidity
rpm. Percentage emulsion was then expressed as: indices (peroxide value, saponification value, iodine value,
% Emulsion = 100 x/y free fatty acids content and acid value) were determined
Where x = height of emulsified layer according to the standard methods of AOAC (2012). The
peroxide values were expressed as miliequivalents of
y = height of whole solution in centrifuge tube.
peroxide oxygen per kg of sample (mEq/kg) while the free
The results were expressed in percentages (g/g basis). fatty acids were expressed as g oleic acid per 100 g of
Determination of anti-nutritional factors sample (g/100 g). The acid value was expressed as mg
NaOH per g of sample (mg NaOH/g). The saponification
The modified method of Reddy et al., (1982) was used for
value was expressed as mg KOH per g of sample (mg
phytic acid and phytate-phospphorus determinations. Phytic
KOH/g). Iodine value was determined by the AOAC (2012)
acid was extracted from each 3 g flour sample with 3%
method using Wij’s iodine solution.
trichloroacetic acid by shaking at room temperature
followed by high-speed centrifugation (30,000 x g for 5 Statistical analysis
min), the phytic acid in the supernatant was precipitated as Data collected from the study were subjected to one-way
ferric phytate and iron in the sample was estimated. Phytate- analysis of variance (ANOVA). Differences among means
phosphorus (phytate-P) was calculated from the iron results were separated using Duncan’s new multiple range test, and
assuming a 4:6 iron: phosphorous molecular ratio (AOAC, significances were accepted at 5% confidence level (P ≤
2012). The phytic acid was estimated by multiplying the 0.05). The statistical software used was SPSS 16.0 for
amount of phytate-phosphorous by the factor 3.55 based on windows.
the empirical formula C6P6O24H18 (Enujiugha and
Olagundoye, 2001).
III. RESULTS AND DISCUSSION
Tannin contents were determined by the modified vanillin-
HCl method (Price et al., 1978). A 2 g sample was extracted Proximate Composition of Parkia biglobosa
with 50 ml 99.9% methanol for 20 min at room temperature Table 1 shows the effects of gamma radiation on the
with constant agitation. After centrifugation for 10 min at proximate compositions of raw and cooked Parkia
653 x g, 5 ml of vanillin-HCl (2% vanillin, 1% HCl) reagent biglobosa seeds. There was an increase in the fat content of
was added to 1 ml aliquots and the colour developed after the cooked seeds after irradiation. This may be due to the
20 min at room temperature was read at 500 nm. Correction rupture of oil cells in the seed skin. Badiani et al. (2002)
for interference by natural pigments in the sample was reported that most nutrients increased their concentration as
achieved by subjecting the extract to the conditions of the a consequence of moisture loss through cooking. The
reaction but without vanillin reagent. A standard curve was protein content of the cooked seeds was reduced by 2.9%
prepared using catechin (Sigma Chemical, St. Louis, MO) from the raw. This is in line with the report of Attia et al.
after correcting for blank, and tannin concentration was (1994) who observed a reduction in protein contents after
expressed in mg/g. cooking chickpea. There was also a slight decrease in the
Determination of oxalate was by the AOAC (2012) method. protein content of seed samples upon irradiation. This might
One gram (1 g) of finely ground sample was dissolved in 75 be due to damage on sulphur-containing amino acids. The
ml of 1.5 N H2SO4. The solution was carefully stirred most radiation sensitive amino acids are those that contain
intermittently with a magnetic stirrer for about 1hr and sulphur, notably cysteine, methionine and tryptophan.
filtered using Whatman no. filter paper. A 25 ml sample of Desulphurization is one of the principal effects of ionizing
the filtrate (extract) was collected and titrated hot (80-90 oC) radiation on amino acids and proteins (Singh et al., 1991).
against 0.1 N KMnO4 solution to the point when a faint pink The single and combined irradiation and cooking treatments
colour appeared that persisted for at least 30 s. The did not significantly (p ≥ 0.05) affect the ash, fibre and
concentration of the oxalate in each sample got from the carbohydrate contents of the locust bean seeds.
calculation: 1 ml 0.1 N permanganate = 0.006303 g oxalate.
All procedures were carried out in triplicates.

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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

Table 1: Proximate composition of raw and treated Parkia biglobosa seeds

Parameters (g/100g) Raw Cooked 5 kGy 10 kGy 10 kGy Irradiated
Irradiated Irradiated and cooked
Moisture Content 4.59a±0.13 4.50a±0.24 4.53a±0.09 4.57a±0.10 4.62a±0.12
Ash content 6.18a±0.02 5.97a±0.03 5.89a±0.03 5.75a±0.02 5.76a±0.04
Total fat/oil 33.92b±0.12 34.06ab±0.10 33.55b±0.11 33.87b±0.09 34.04a±0.12
Crude Fibre content 5.57a±0.02 5.24a±0.01 5.82a±0.02 5.60a±0.02 5.71a±0.02
Crude protein 32.82a±0.14 31.88b±0.12 32.73a±0.13 32.48a±0.13 32.22ab±0.12
Carbohydrate content 20.45a±0.32 20.56a±0.24 21.57a±0.29 21.47a±0.26 21.41a±0.42
Gross Energy (KJ 100g-1 DM) 2168.393 2212.59 2171.645 2165.8 2250.559
*Values are means of triplicate determinations (n=3) ± standard deviation. Values along same row with the same letters in
superscript are not significantly different (p≥0.05).

Table 2: Amino acid composition of raw and treated P. biglobosa seeds

Parameters Raw Cooked Irradiated(10kGy) Irradiated+Cooked
Lysine* 3.54c±0.01 3.60b±0.02 3.76a±0.01 3.19d±0.04
Histidine* 2.29c±0.12 2.54b±0.02 2.57a±0.03 2.07d±0.02
Arginine* 10.03b±0.04 10.04b±0.03 10.30a±0.03 9.36c±0.22
Asparagine 10.60a±0.03 10.15c±0.04 10.59a±0.02 10.28b±0.14
Threonine* 3.27a±0.02 3.00b±0.02 3.27a±0.01 3.27a±0.02
Serine 2.91b±0.12 2.55c±0.03 3.05a±0.02 2.54c±0.03
Gluthamine 16.20a±0.04 14.14c±0.21 15.14b±0.30 13.24d±0.02
Proline 3.29c±0.02 4.14b±0.02 4.46a±0.01 2.97d±0.01
Glycine 4.69b±1.02 4.52c±0.13 5.01a±0.13 4.17d±0.20
Alanine 3.94b±0.03 3.90c±0.72 4.05a±0.42 3.59d±1.33
Cysteine* 1.65c±0.22 1.69b±0.66 1.72a±0.92 1.39d±0.12
Valine* 3.83c±0.14 5.08a±0.13 5.00b±0.34 3.60d±0.87
Methionine* 1.41a±0.26 1.30c±0.01 1.38b±0.12 1.20d±0.01
Isoleucine* 3.95b±0.03 3.83c±0.05 4.14a±0.03 3.39d±0.18
Leucine* 6.05a±0.01 5.66c±0.01 5.85b±0.01 5.02d±0.01
Tyrosine* 1.61d±0.00 2.74b±0.02 2.90a±0.04 2.25c±0.02
Phenyalanine* 4.56b±0.02 4.31c±0.18 4.65a±0.14 3.97d±0.06
Tryptophan 2.10a±0.05 1.97a±0.02 2.08a±0.07 1.95a±0.01
*Values are means of triplicate determinations (n=3) ± standard deviation. Values along same row with the same letters in
superscript are not significantly different (p≥0.05).

Effects of processing on the amino acid profile of Parkia enzymes, hormones and other metabolic molecules (Olotu
biglobosa et al., 2014b). Glutamate and Aspartate accounted for about
Table 2 presents the results of the effect of processing 30% of the amino acids in the sample which is similar to
(cooking, irradiation and a combination of both) on the amino acid profiles reported previously in some oil seeds
amino acids composition of Parkia biglobosa seeds. Amino (Olotu et al., 2014b; Igwe et al., 2012). Sulphur-containing
acids are needed for the synthesis of most body tissues, amino acids (methionine and cysteine) were the least

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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

concentrated with values ranging from 1.20 g to 1.41 g and (Bamidele, 2015). This finding is in line with the report of
1.39 g to 1.72 g, respectively. The reduced concentration of Abdel-Ghaffar (2013), who reported an increase in some
methionine could be attributed to the simple amino acids amino acids of soy flour.
undergoing reductive deamination and decarboxylation Effect of processing on the mineral composition of
during irradiation (Enujiugha et al., 2023a). Cooking Parkia biglobosa
significantly (p ≤ 0.05) reduced the amino acid contents
The results of the mineral analysis as shown in Table 3
except for valine which is heat stable. The reductive effects
indicate that the locust bean seeds were richer in iron and
of cooking on protein and amino acid compositions of the
potassium after cooking, which could be interpreted as a
seeds may be attributed to Amadori rearrangements that
consequence of the probable inactivation of anti-nutrients.
may go beyond the deoxy-ketosyl stage. It may also be due
Sodium was lost in the cooked as well as post-irradiated
to the formation of D-amino acids which results from high
cooked samples. This may be due to its solubility and its
and prolonged heat treatment (Olaofe et al., 1994). This was
ability to leach-off in cooking water. It can however be
likely the case because the method used for amino acid
stated from the findings that mineral bioavailability can be
analysis will only detect L-amino acids from animal and
increased with cooking with little impact on the mineral
plant proteins that do not produce racemisation (Adeyeye et
constituent of the seed. Zinc was significantly increased in
al., 2010). Increase in the amino acid concentration of the
cooked as well as 10 kGy irradiated samples
10 kGy irradiated sample was observed with arginine,
histidine, tyrosine, glycine, lysine, proline, threonine, These minerals act as stabilizers of the structures of
phenylalanine and serine. The increase of alanine, glutamic membranes and cellular components. Zinc is an important
acid, valine, methionine, isoleucine and cystine were component of several enzymes and their biochemical
significant (p > 0.05). The changes in the concentration of functions, especially in the synthesis and degradation of
amino acids induced by irradiation may be due to free macromolecules such as carbohydrates, proteins, lipids and
radicals of the peptide bonds, deamination and nucleic acids as well as wound healing (Frossard et al.,
decarboxylation reactions of amino acids followed by 2000).
chains of chemical reactions forming other new radicals
Table 3: Mineral composition of raw and treated Parkia biglobosa seeds
Parameters Raw Cooked 5 kGy Irradiated 10 kGy 10 kGy Irradiated and
(mg/100g) Irradiated cooked
Potassium 18.21a±1.65 14.47b±0.65 15.25b±0.15 17.30a±0.65 15.57b±0.15
Sodium 2.88a±0.10 2.68ab±0.07 2.57b±0.18 2.54b±0.10 2.18c±0.13
Calcium 14.68c±0.55 18.30a±1.08 18.17b±0.21 15.86c±1.91 11.18d±0.19
Magnesium 4.61a±0.82 3.49b±0.24 3.40b±0.10 3.60b±0.14 2.85b±0.25
Zinc 4.19c±0.02 5.19b±0.03 4.59b±0.75 6.34a±0.06 4.19c±0.59
Iron 1.46b±0.02 1.73a±0.04 1.42bc±0.11 1.24cd±0.20 1.09d±0.03
Phosphorus 3.67bc±0.39 5.75a±0.03 4.25b±0.06 4.00b±0.55 3.36c±0.18
*Values are means of triplicate determinations (n=3) ± standard deviation. Values along same row with the same letters in
superscript are not significantly different (p≥0.05).

Effect of processing on Functional properties seed flour. Dissociation and denaturation results in
The effects of cooking, irradiation and combined treatment increased oil absorption of treated proteins compared to
of both on the functional properties of locust bean seeds are native proteins (Siddharaju et al., 2002). This observation is
presented in Table 4. Water absorption capacity (WAC) is consistent with the results of Sosulski et al. (1976) who
an important functional property of proteins and is a reported similar results using sunflower seeds.
measure of the quality (juiciness, texture, binding of Cooking significantly reduced the foaming capacity by
structure, appearance and mouth feel) of flour. There was a about 92.6%. This finding is in agreement with that reported
significant decrease in the water absorption capacities of the by Yusuf et al. (2008) for Bambara groundnut which was
cooked (as a single treatment) sample and the irradiated attributed to protein denaturation. Irradiation at 10 kGy
cooked sample compared to the raw. Cooking led to decreased the foaming capacity of the seed flour
significant increase in oil absorption capacity (OAC) of the significantly by 25.9%, probably owing to extensive
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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

denaturation and protein cross-linking. However, given the Pentaclethra macrophylla (Enujiugha et al., 2012) and
foaming capacity values in the present study, African locust groundnut (Enujiugha et al., 2023a).
bean seed flour has its foaming capacity compromised by Emulsion capacity denotes the maximum amount of oil that
all the treatments. The 5 kGy irradiated sample has the can be emulsified by protein dispersion. The high emulsion
highest foam stability while the cooked sample has the capacity in the present study could be as a result of high
lowest foam stability. This agrees with the findings of Lin content of free fatty acids which leads to increased oil
et al. (1974) which stated that native proteins give higher absorption (Ihekoronye and Ngoddy, 1985). Irradiation at 5
foam stability than denatured protein. kGy caused a significant decrease in the emulsion capacity
Significant reduction in least gelation concentration (LGC) of the seed flour accounting for more than 10% loss of
was observed at 5 kGy gamma irradiation dose (p < 0.05). emulsion capacity. Irradiation at 10 kGy had no significant
Such a decrease in LGC might be attributed to increased (p<0.05) effect on emulsion capacity of the oil seed flour
interaction of proteins with water (Adebowale and Lawal, when compared with the non-irradiated (control) samples.
2004). Improvement in gelation property is beneficial as it The decrease in emulsion capacity has been speculated to
allows the utility of seed flour in preparation of food have resulted from changes, such as protein aggregation as
products like custards, ice creams, sausages and other well as surface hydrophobicity and charge characteristics
bakery products (Bhat and Sridhar, 2008). The current (Cheftel et al., 1985).
findings are in agreement with results obtained for
Table 4: Functional properties of raw and processed Parkia biglobosa seeds
Parameters Raw Cooked 5 kGy 10 kGy 10 kGy Irradiated and
Irradiated Irradiated cooked
WAC (ml/g) 380.00a±0.21 360.00ab±0.35 360.00b±0.08 380.00a±0.02 360.00ab±0.03
LGC (m/v) 8.00a±0.01 6.00a±0.02 2.00b±0.03 4.00b±0.01 2.00b±0.02
OAC(ml/g) 190.00ab±0.03 210.00a±0.02 175.00b±0.02 180.00b±0.01 200.00ab±0.02
FC (%) 45.00a±0.03 3.33e±0.12 33.33b±0.02 25.00c±0.04 6.67d±1.23
EC (%) 380.00a±0.09 360.00b±0.01 340.00c±0.31 380.00a±0.03 360.00b±0.01
*Values are means of triplicate determinations (n=3) ± standard deviation. Values along same row with the same letters in
superscript are not significantly different (p≥0.05).
WAC = Water absorption capacity; OAC = Oil absorption capacity; LGC = Least gelation concentration; FC = Foaming
capacity; EC = Emulsion capacity

Effect of processing on anti-nutritional factors irradiation alone in order to keep oxalate intake as low as
The levels of some anti-nutritional components in the raw possible. Observations were comparable to the reports of
and processed seed flour samples are presented in Table 5. Albihn and Savage (2001) and Wanasundera and Ravindran
Anti-nutritional factors are generally reported to have the (1992). The reduction in oxalate levels on cooking is to
effect of lowering digestibility and absorption of important enhances the bioavailability of essential dietary nutrients of
dietary nutrients. the African locust bean seeds

Oxalate can have deleterious effects on human nutrition and The results obtained indicate that phytate content decreased
health, particularly by decreasing calcium absorption and slightly with treatment. Irradiation alone effectively
aiding the formation of kidney stones (Noonan and Savage, reduced the phytate level in the oil seed by 4.8% compared
1999). Cooking has been found effective in the reduction of to the control. Decrease in the phytate content in the
oxalate content in oil seeds (Chai and Liebman, 2005). A irradiated and cooked sample was as high 24.8%. The
decrease in the oxalate content was observed in both cooked reduction in phytate content during cooking may be partly
samples. The higher percentage of oxalate reduction occurs due to the formation of insoluble complexes between
during boiling and may be due to its solubility in water. phytates and other components, such as phytate-protein and
Boiling may cause skin rupture and facilitate the leakage of phytate-protein-mineral complexes, or to the inositol
soluble oxalate from seeds into cooking water (Albihn and hexaphosphate being hydrolyzed (Siddhuraju and Becker,
Savage, 2001). Comparing the different treatments, cooking 2001). Cooking has been reported to lower the phytate
and post-irradiation cooking are to be preferred to levels in several plant foodstuffs (Badifu, 2001).
Taghinejad-Roudbaneh, (2010) in a study where canola
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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

meal was irradiated using electron beams up to 45 kGy dose complexes with proteins thereby interfering with their
observed a total disappearance of phytic acid at 30 kGy and bioavailability (Enujiugha and Agbede, 2000). Reduction of
a reduction of up to 89.66% at 15 kGy irradiation dose. tannins might have resulted in a significant increase in Zinc
Currently there is evidence that dietary phytate at low levels content at 10kGy irradiation dose.
may have beneficial effect as an antioxidant, anticarcinogen Rancidity indices of oil from Parkia biglobosa
and may likely play an important role in the control of
The products of rancidity are known to be hazardous to
hypercholesterolemia and atherosclerosis (Kumar et al.,
health since they are associated with aging, membrane
2010; Phillippy et al., 2004).
damage, heart disease and cancer (Cosgrove et al., 1987).
The high tannin contents of the seed might have been a The rancidity indices of oil as affected by processing are
major cause of the astringency observed when tasted in its presented in Table 6.
unprocessed form. Tannins usually form insoluble
Table 5: Anti-nutritional factors in raw and treated Parkia biglobosa seeds
Parameters Raw Cooked 5 kGy Irradiated 10 kGy 10 kGy Irradiated and
Irradiated cooked
Oxalate(mg/g) 0.15a±0.02 0.06±d0.13 0.09c±1.04 0.11b±0.18 0.06d±0.05
Phytate(mg/g) 1.25b±0.02 1.19a±0.02 1.57a±0.02 1.11b±0.02 0.94b±0.03
Tannin(mg/g) 0.02a±0.01 0.02a±0.01 0.02a±0.01 0.02a±0.02 0.02b±0.01
Phytate-P 0.35b±0.12 0.34bc±0.05 0.44a±0.04 0.31bc±0.01 0.33c±0.26
*Values are means of triplicate determinations (n=3) ± standard deviation. Values along same row with the same letters in
superscript are not significantly different (p≥0.05).

Table 6: Rancidity indices of the raw and treated seed oil extract
Parameters Raw Cooked 5 kGy 10 kGy 10 kGy Irradiated and
Irradiated Irradiated cooked
Peroxide value (mEq./kg) 0.71c±0.05 0.73c±0.10 0.73c±0.28 0.79b±0.05 0.86a±0.06
Iodine Value (mg/100g) 140.13a±1.80 133.53b±2.82 142.48a±0.17 144.40a±2.05 122.77b±8.85
Acid Value (mgNaOH/g) 1.07c±0.03 1.11ab±0.01 1.07c±0.01 1.09bc±0.02 1.14a±0.02
Saponification Value 176.33c±0.21 184.00b±0.10 173.33b±0.01 180.67b±0.02 186.67a±0.09
(mgKOH/g)
Free Fatty Acid 1.31cd±5.51 1.62ab±2.00 1.63d±2.08 1.69c±1.15 1.85a±1.53
(% Oleic acid)
*Values are means of triplicate determinations (n=3) ± standard deviation. Values along same row with the same letters in
superscript are not significantly different (p≥0.05).

The peroxide value is an indicator of oxidative rancidity. As peroxide value of the oil while 5kGy irradiation dose
oxidation takes place, the double bonds in the unsaturated resulted in 2.82% increase in the peroxide value. This
fatty acids are attacked leading to the formation of increase conforms to the work of Farag et al. (1992) who
peroxides. Fresh oils have been shown to have peroxide reported acceleration of cottonseed oil oxidation during
values lower than 10 mg/g and oils become rancid with irradiation heating which as indicated by an increase in
peroxide value ranging from 20 to 40 mg/g (Oyinloye and peroxide value. Increases in peroxide values are expected as
Enujiugha, 2017). The highest peroxide value was observed irradiation produces large amounts of free radicals that
in the oil from the seeds with combined treatments of enhances lipid peroxidation (Sajilata and Singhal, 2006).
irradiation and cooking. These results are generally in Iodine value is a measure of the unsaturation of fats and oils.
agreement with those of Mexis et al. (2009) for gamma It is based on the ability of an unsaturated carbon to carbon
irradiated almonds where the peroxide values increased bond to add halogen atoms. Irradiation treatments alone (5
with dosage. Cooking alone accounted for 3.6% increase in kGy and 10 kGy) did not significantly affect the iodine

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Asunni et al. Amino Acid Profile and Physicochemical Properties of African Locust Bean (Parkia biglobosa) Seeds
as affected by Combined Irradiation and Cooking

value of the resultant oil. However, significant decreases in ETHICAL APPROVAL AND CONSENT TO
iodine value were observed for the cooked and post- PARTICIPATE
irradiated cooked samples (133.53 g I2 per 100 g and 122.77 This work is part of a wider research which was approved
g I2 per 100 g respectively). These were in agreement with by the Ethics Committee of the School of Agriculture and
the reports of Omafuvbe et al., (2004) who observed a Agricultural Technology, Federal University of Technology
decrease in iodine value in African Oil Bean seeds as a with assigned number FUTA/SAAT/ETH/2011/14
result of cooking.
The acid value of oils indicates the total acidity as estimated
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