Nigerian Journal of Animal Science and Technology

Nig. J. Anim. Sci. Tech. Vol. 4 (1):43 - 52

Available @ www.njast.com.ng
© Department of Animal Science, University of Maiduguri
Chemical composition, anti-nutritive substances, amino acid profile and mineral
composition of Moringa oleifera seeds subjected to different boiling duration
1
Musa, J. and 2*Njidda, A. A.
1
Department of Animal Science, Faculty of Agriculture, Kaduna State University, Kafanchan campus, Kaduna
State, Nigeria
2
Department of Animal Science and Fisheries, National Open University of Nigeria, KM4, Kaduna-Zaria road,
Rigachukun, Kaduna State, Nigeria
*Corresponding Author’s email:ahmednjidda7@gmail.com

Abstract
This study was conducted to evaluate the chemical and mineral compositions, anti-nutritive
substances and amino acid profile of Moringa oleifera seeds. Samples were collected from
five Moringa oleifera trees and analyzed for chemical and mineral compositions, anti-
nutritive substances and amino acid profile using standard methods. The results indicated
that the seeds of Moringa oleifera contained significantly (p<0.05) higher (322.00 and
136.40 g kg-1 DM) crude protein (CP) and fat values. The highest CP (322 g kg-1 DM) was
obtained at 60 minutes boiling point of M. oleifera while the lowest CP (225.00 g kg-1 DM)
was obtained from M. oleifera seeds boiled at 0 minutes (no heat treatment). M. oleifera
seeds was observed to contain significantly (p<0.05) higher crude fiber (94.00 g kg-1 DM),
neutral detergent fiber (518.00 g kg-1 DM) and cellulose (338.00 g kg-1 DM) at 60 minutes
boiling duration than the other parameters studied. Boiling duration shows significant effect
(p<0.05) on Sodium, potassium, calcium and magnesium values which decrease with
increase in boiling duration while the phosphorus shows some inconsistency in the values
with variation in boiling time. Similarly the values for the micro minerals tend to decrease
with increase in boiling time but the changes was so insignificant (figuratively). The tannin,
phenolics, saponins and phytate content was observed to decrease with increase in boiling
time. In conclusion, the seeds of both Moringa oleifera could be used as alternative protein
supplement sources for feeding ruminant and monogastric animals during dry periods of the
year and apparently boiling duration (60 minutes boiling duration) reduces the level of anti-
nutrtive substances and macro minerals, as such mineral supplement should be added when
using heat treated M. oleifera seeds does not cause adverse effects on the nutrient
composition of the seeds.
Keywords: Moringa oleifera seeds, mineral, anti-nutritive substances

Introduction lotion. People in the Indian subcontinent

Moringa is considered to be one of the most
nutrient-rich plant on earth. Moringa leaves
have been consumed by Asians for
millennia as a healthy food product. Studies
from other countries indicate that the leaves
have immense nutritional value such as
phytochemicals, vitamins, minerals, and
amino acids (Busani et al., 2011). As such,
the leaves have been used to combat
malnutrition, especially among infants and
nursing mothers. The Romans, Greeks, and
Egyptians extracted edible oil from the
seeds and used it for perfume and skin
have long used Moringa pods for food. The
leaves are consumed throughout West
Africa and some parts of Asia. Moringa
leaves are of high nutritive value (Moyo et
al., 2011). Moringa oleifera belongs to the
family Moringaceae and it is an effective
remedy for malnutrition. Moringa is rich in
nutrition owing to the presence of a variety
of essential phytochemicals present in its
leaves, pods and seeds. In fact, Moringa is
said to provide 7 times more vitamin C than
oranges, 10 times more vitamin A than
carrots, 17 times more calcium than milk, 9
 Musa and Njidda 2021
times more protein than yoghurt, 15 times
more potassium than bananas and 25 times
more iron than spinach (Rockwood et al.,
2013). Moringa seeds have long been used
by the public as a tasty vegetable and water
purifier because of its coagulant properties
(Ayotunde et al., 2011). Moringa oleifera is
a promising plants with organic properties
which could contribute to increased intake
of some essential nutrients and health-
promoting phytochemicals (Balbir, 2006). It
contains active substances that can improve
digestion and metabolism and possess
antibacterial and immuno stimulant
activities (Ghazalah and Ali, 2008). It also
contains bioceutical agents that could
substitute synthetic growth enhancers and
supplements in poultry birds (Lannaon,
2007). Other Moringa plant parts like
flowers, roots, and bark also have good
nutritional and therapeutic value (Olushola,
2006). Seed extract exerts its protective
effect by decreasing liver lipid peroxides,
antihypertensive compounds thiocarbamate
and isothiocyanate glycosids have been
isolated from the acetate phase of the
ethanolic extract of Moringa pods.
However, there is a paucity of information
on the effect of boiling duration on seeds of
M. oleifera cultivated in Nigeria for its
inclusion in livestock feed formulation.
Therefore, this study was conducted to
analyze the chemical composition, anti-
nutritive substances, amino acid profile and
mineral composition of M. oleifera seeds
meal with an objective of addressing
food/feed availability for livestock in
Nigeria
Materials and Methods
Forage samples collection and
experimental site:
The seeds of Moringa oliefera were
harvested from at least 10 different Moringa
oleifera stands selected at random in four
locations within the study area at the end of
the rainy season. The harvested sample were
then pooled and then oven dried at 1050C
for 24h to constant weight and ground to
pass through a 1.0 mm, sieve. The samples
44
were then sub-sampled to obtain three
samples and used for the laboratory
analysis.
All Moringa oliefera seeds were harvested
from Kafanchan, Jema’a local Government
area of Kaduna state, Nigeria. It is located
within latitude 90 34’N and longitude 80
17’E. The vegetation of the area is the
Guinea Savannah type and the area is
designated as koppen’s Aw climate with two
distinct seasons, a wet season in summer
and a dry season in winter. Rainfall occurs
between the months of April to October
with a peak in August. The mean annual
rainfall is about 1800 mm and the mean
monthly temperature is 250C, while the
relative humidity is about 630C, (Ishaya and
Abaje, 2008).
Chemical Analysis:
Samples of Moringa oleifera seeds were
analyzed for dry matter (DM), crude protein
(CP), ether extract (EE), crude fibre (CF)
and ash according to AOAC (2002). The
mineral composition (calcium and
magnesium) of the forages were analyzed
using atomic absorption spectrophotometry
(Zohary, 1973). Phosphorus was determined
according to the Vanadomolybdophosphoric
Acid Method (Shiou, 1996) using a
spectrophotometer (Jenway 6100, UK). The
seed were analyzed for neutral detergent
fibre (NDF), acid detergent fibre (ADF),
acid detergent lignin (ADL) and cellulose
according to Van Soest et al. (1991).
Percentage hemi-cellulose content was
obtained by finding the difference between
NDF and ADF values. Anti nutritional
constituents: Phytate in the plant seeds was
estimated as phytic acid using the method of
Maga (1982), while saponins and total
condensed tannin were determined as
reported by Babayemi et al., (2004) and
Polshettiwar et al., (2007), phenolics was
determine using Folin Ciocalteu as
described by Makkar (2000).
Statistical analysis
 Musa and Njidda 2021
The data were analyzed using SPSS (2009).
The mean and standard error of means
(SEM) of the triplicate analyses of the
samples were calculated. Analysis of
variance was performed to determine
significant differences between the means,
while the means were separated using the
new Duncan multiple range test and P <
0.05 was applied to establish significant
differences. Determination of
Determination of amino acid profile
Moringa seed flour sample hydrolysates
were prepared following the method of
Spackman et al. (1958). 200 mg of samples
was weighed into a glass ampoule, 5 mL of
6 mol/L HCl was added to this, and the
contents were hydrolyzed in an oven preset
at 105±5°C for 22 h. Oxygen was expelled
in the ampoule by passing nitrogen gas into
it. Amino acid analysis was done by ion-
exchange chromatography (Spackman et al.
1958) using a Technicon Sequential
Multisample Amino Acid Analyzer
(Technicon Instruments Corporation, New
York, NY). The period of analysis was 76
min, with a gas flow rate of 0.50 mL/min at
60°C, and the reproducibility was ±3%. The
amino acid composition was calculated from
the areas of standards obtained from the
integrator and expressed as percentages of
the total protein.
Results
Chemical composition of test sample
The result of the chemical composition of
the test sample is presented in Table 1. M.
oleifera seeds contained significantly
(p<0.05) higher values for all the parameters
observed except for crude fibre and
hemicellulose which tends to be low for all
the treatments. Significant differences
(P<0.05) were observed between the boiling
duration for all the parameters studied. The
results also reveal an increase in values with
increase in boiling duration for all the
parameters studied. The results also revealed
an increase in values with increase in
boiling duration for dry matter (DM),
organic matter (OM), crude protein (CP),
45
ether extract (EE) and crude fibre (CF)
while the values for all the fibre fractions
neutral detergent fibre (NDF), acid detergent
fibre (ADF), acid detergent lignin (ADL)
and Cellulose, decrease with increase in
boiling duration. Ash and hemicellulose
were inconsistent (as shown in Table 1).
Macro minerals concentration in M.
oleifera seed
The result of the macro mineral
concentration is shown in Table 2. It was
observed that the values for all the macro
minerals tends to decrease with increase in
boiling duration. There was significant
difference (P<0.05) for all the parameters
studied. The decrease in values of the
mineral elements occur mostly at 40 minutes
boiling duration.
Micro minerals concentration in M.
oleifera seed
Table 2 presented the composition of micro
minerals in the M. oleifera seeds in this
study. The iron content of the seeds ranged
between 183.00 and 185.00 mg/100g DM.
Boiling duration had significant effect
(P<0.05) on most micro minerals except for
Manganese (P>0.05). The concentration of
copper in the M. oleifera seeds was
generally low and non-significant for all the
boiling duration (7.42 to 7.60 mg/100g)
Amino Acid profile
The result of the amino acid profile is shown
in Table 3. Threonine valine, isolencine,
lysine, histidine, phenylalamine and
Tyrosine were observed to increase with
increase in boiling durations. There was
significant difference (P>0.05) between the
boiling duration for all the amino acids. The
non-essential amino acids also differed
significantly (P>0.05) with boiling duration.
The values tend to increase with increase in
boiling duration.
Anti-nutritive Substances
All anti-nutritive substances (Table 4) were
significantly (p>0.05) affected by boiling
duration with 60 minutes boiling time
 Musa and Njidda 2021
having the least value and 0 minutes having
the highest. Thus boiling reduces the anti-
nutritive substances in the seeds.
Discussion
Chemical composition Moringa oleifera
seeds
Moringa oleifera tree contains high crude
protein (CP) in the leaves (251 g/kg DM)
and negligible content of tannins and other
anti-nutritive compounds and offers an
alternative source of protein to ruminants
(Nouala et al., 2006; Raji and Njidda, 2014)
and non-ruminants. The seeds contain high
amount of CP, suggesting that M. oleifera
can be used as a protein source for both
livestock and humans. The fact that the
seeds contain higher CP content than other
parts suggests that they can be used as a
valuable source of protein. Ojukwu (2012)
stated that Moringa seed are periodically
harvested to make a sauce, locally known as
“mboum” or can be used to feed poultry,
pigs and cattle. In a recent study, Mabruk et
al. (2010) reported higher levels of DM, ash,
CP, EE and CF (950.00, 34.80, 391.70,
388.00 and 48.00 g/Kg) while gross energy
was (14.790) (MJ/kg). These results
indicated that nutrient composition of M.
oleifera seeds differed according to location
and possibly stage of harvesting of Moringa
seeds. M. oleifera seed is a good source of
protein since according to Pearson (1976),
any plant food that provides more than 12%
of its calorific value from protein is a good
source of protein. The seeds contain high
amount of proteins at higher level of boiling
time which indicate that the boiling duration
has no negative effect on the CP content of
the M. oleifera seeds. M oleifera is also a
good source of lipid (33.78%), and
carbohydrates (28.77%) with a calorific
energy value of 531.18 Kcal/100g.
Minerals of Moringa oleifera seeds
Moringa oleifera contains a considerable
amount of various nutrients, and has been
suggested as a good supplement for such
nutrients as protein, fibre and minerals
(Jongrungruangchok et al., 2010). Several
46
works have been done, reporting the levels
of nutrients, including minerals in M.
oleifera, with emphasis on variations in the
levels as a function of geographical
locations where the plant is grown; and the
results showed that there could be variations
in the levels of macro and micro mineral
elements in the plant, depending on the soil
properties of the geographical locations
where it grows (Anjorin et al., 2010) and
Njidda, (2012). Generally, the results
showed significant effect (p<0.05) of boiling
duration on the macro minerals, the values
of the macro minerals tend to decease with
increase in the boiling duration which
suggest that adequate mineral
supplementation is required when feeding
heat treated M. oleifera seeds to livestock.
The vital minerals present in Moringa
include Calcium, Copper, Iron, Potassium,
Magnesium, Manganese and Zinc. The
contents of sodium (Na), potassium (K),
cacium (Ca), phosphorous (P) and
magnesium (Mg) and trace minerals iron
Fe), copper (Cu), Manganese and zinc (Zn)
were also higher.
Amino Acids Moringa oleifera seeds
Amino acids are classified as essential and
non-essential, which vary according to
animal species and their production system
(Swanepoel et al., 2010). Rumen microbes
synthesize the essential amino acids from
other amino acids or from nitrogen
containing substances (Moyo et al., 2011).
The efficiency of rumen microbial growth
and activity in the rumen is enhanced by the
presence of adequate amino acids, peptides
and most macro and micro minerals
(Swanepoel et al., 2010). Each amino acid
has a specific function in the animal’s body.
In general, amino acids are required for the
production of enzymes, immunoglobins,
hormones, growth, repair of body tissues
and form the Amino acids also affect the
function of other nutrients in the animal’s
body such as presence of lysine, which
ensures adequate calcium absorption and
aids in the antibody production (Brisibe et
al., 2009). In addition, they contribute to the
formation of glucose, acting as a buffer
 Musa and Njidda 2021
when other precursors are in short supply
(Swanepoel et al., 2010).
Anti-nutritional factors of Moringa
oleifera seeds
Some of the anti-nutritional factors present
in the Moringa oleifera seed include
Tannins, Phytates and Oxalates, Saponins
and Cyanogenic glycosides among others.
Tannins (40 mg/100g) and oxalates (51.24
mg/100g) are very high in the seeds of
Moringa oleifera. The values for phenolics,
saponins and phytates are considerably
lower, 0.00 to 40.00 1.00 to 9.00 mg/100g
and 0.00 to 70.00 mg/100g respectively.
Tannins have antagonistic competition with
proteins, consequently lowering the bio-
availability of protein. Tannins can lower
the available proteins by antagonistic
competition and can therefore elicit protein
deficiency. It can also prevent protein
digestion by inhibiting the activation of
pepsinogen and chymotrypsinogen to pepsin
and chymotrypsin respectively (Njidda and
Ikhimioya, 2012). Phytic acid have
complicated effects in animal system,
particularly indigestion of food and
flatulence (Akubugwo et al., 2007). Phytic
acid and oxalates have complicated effects
in the human system including reacting
tightly with divalent ions such as calcium
and zinc ions, thereby making them
unavailable to the body (Akubugwo et al.,
2007). However, these anti-nutrients can be
removed through soaking, boiling or even
frying (Igwilo et al., 2007). The saponin
content of moringa oleifera sample was
lower when compared with that of Moringa
oleifera (5.20 mg/100 g) reported by Seena
(2006) and tend to decrease with increase in
boiling duration. It is evident that anti-
nutrients and phytochemicals have both
adverse and beneficial effects in humans
(Njidda, 2011; Soladoye and Chukwuma
2012). For example, phytic acid, lectins,
phenolic compounds and tannins, saponins,
enzyme inhibitors, cyanogenic glycosides,
and glucosinolates reduce the bioavailability
of certain nutrients and impair growth in
children (Elemo et al. 2001; Dingynan et al.
47
2003). On the contrary, when phytic acid,
lectins, and phenolic compounds and
saponins were used at low levels, they
exhibited hypoglycemic,
hypocholesterolemic and anticancer
properties (Jariwalla et al. 1990; Oakenfull
and Sidhu 1990). Flavonoids have also been
reported to possess antioxidant, anti-
inflammatory, and antihypertensive
properties (Ayinde et al. 2007; Li-Weber
2009).
Conclusion
In conclusion, this study established boiling
moringa oleifera seed upto 60 minutes
duration have significant effect in reducing
the anti-nutritive substances and the macro
mineral contents of the seed, as such
supplemental macro minerals should be
taking into consideration when using heat
treated M. oleifera seed in formulating diet
for livestock. Boiling duration (60 minutes)
does not show any negative effect on the
amino acids profile studied.
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Table 1: Effect of different boiling duration on the proximate composition of Moringa oliefera seeds (g kg-1 DM)
Boiling duration in minutes
Parameters 0 10 20 30 40 50 60
b b b a a
Dry matter 916.00±1.92 916.50±2.79 917.00±1.09 918.80±1.23 920.00±1.45 922.00±1.32a 926.00±1.65a
Ash 45.00±0.29a 44.00±0.52a 42.00±0.22a 40.00±0.32a 38.00±0.12ab 36.00±0.12b 34.00±0.02b
c c c c b
Organic matter 871.00±1.34 872.50±1.46 875.00±1.46 878.80±1.56 882.00±1.21 886.00±1.12b 892.00±1.33a
c c c b b
Crude protein 225.00±0.56 226.00±0.77 229.00±0.92 303.80±0.56 307.00±0.33 309.00±0.23b 322.00±0.72a
a a a a a
Ether extract 130.00±0.55 131.00±0.23 133.00±0.33 134.00±0.26 134.50±0.37 135.00±0.24a 136.40±0.23a
Crude fibre 86.00±0.56c 87.00±0.11b 88,00±0.21b 88.50±0.12b 89.00±0.43b 92.00±0.24a 94.00±0.27a
a b b c c
Carbohydrate 430.00±1.23 428.50±0.96 425.00±0.94 352.50±0.56 351.50±0.23 350.00±0.45c 339.60±0.34d
a b c c d
Neutral Detergent Fibre 544.00±1.56 540.00±1.34 538.00±1.21 530.00±0.46 527.00±0.76 521.00±0.66e 518.00±0.43f
a b c d e
Acid Detergent Fibre 500.00±1.33 490.00±1.23 487.00±1.23 482.00±1.67 478.00±1.28 475.00±1.23f 472.00±1.11g
a b c d e
Acid Detergent Lignin 152.00±0.56 150.00±0.27 146.00±0.57 142.00±0.55 139.00±0.09 137.00±0.57f 134.00±0.23g
Cellulose 348.00±1.56a 340.00±1.37c 341.00±1.27b 340.00±0.21c 339.00±0.56d 338.00±0.23e 338.00±0.22e
d a a c b
Hemicellulose 44.00±1.42 50.00±1.32 51.00±1.43 48.00±1.33 49.00±1.33 46.00±1.13c 46.00±1.23c
a, b, c, d= means within the same row bearing different superscript are significantly different (p<0.05)
Table 2: Effect of different boiling duration on mineral composition Moringa oliefera seeds
Boiling duration in minutes
Parameters 0 10 20 30 40 50 60
Macro minerals (mg/100g)
Sodium 1470.00±1.26a 1468.00±1.12b 1466.00±1.34c 1460.00±1.32d 1455.00±1.54e 1450.00±1.55f 1448.00±1.23g
Potassium 1273.00±1.56a 1272.00±0.27b 1270.00±1.23c 1268.00±1.23d 1266.00±1.23e 1264.00±1.23f 1261.00±1.23g
Calcium 254.00±2.56a 254.00±1.23a 253.00±1.11b 253.80±1.09b 253.50±1.03b 253.30±1.03b 252.80±1.23c
a a ab bc
Phosphorus 738.00±2.33 738.00±2.13 737.00±1.23 736.00±1.44 736.00±1.62bc 736.00±1.26bc 730.00±1.92d
Magnesium 64.20±0.56a 64.18±0.27a 64.10±0.34a 64.00±0.78a 63.50±0.56b 63.20±0.96b 63.00±0.29b
Micro minerals (mg/100g)
Iron 185.20±1.33a 185.00±1.33a 184.60±1.24b 184.600±1.56b 184.50±1.11b 184.00±1.24b 183.000±1.24c
a a
Copper 7.60±0.56 7.60.65 7.54±0.12b 7.50±0.16b 7.47±0.12b 7.45±0.17c 7.42±0.12c
Manganese 91.60±2.23a 91.60±0.89a 91.60±0.56a 91.58±0.77a 91.50±0.23a 91.46±1.27a 91.40±0.12a
a a a a b
Zinc 57.30±1.33 57.30±1.56 57.20±1.10 57.00±1.126 56.80±0.56 56.50±1.61b 56.00±1.02b
a, b, c, d= means within the same row bearing different superscript are significantly different (p<0.05)

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Table 3: Effect of different boiling duration on amino acid profile of Moringa oliefera seeds
Boiling duration in minutes
Parameters 0 10 20 30 40 50 60
Non-essential amino acids (mg/100g)
Threonine 920.00±2.23f 920.00±2.26f 922.00±2.11e 924.00±1.26d 926.00±3.26c 928.00±2.17b 930.00±2.44a
Valine 1440.00±2.26e 1440.00±2.11e 1443.00±2.01d 1447.00±2.17c 1450.00±2.26b 1452.00±2.26a 1452.00±1.22a
Isoleucine 1232.00±1.33e 1233.00±1.26d 1235.00±1.13c 1238.00±2.26d 1241.00±2.26c 1245.00±2.26b 1250.00±1.12a
Leucine 2080.00±2.22g 2081.00±2.05f 2083.00±2.18e 2085.00±2.43d 2088.00±2.11c 2090.00±2.13b 2100.00±3.24a
Lysine 610.00±3.26g 613.00±2.72f 616.00±3.11e 620.00±1.27d 623.00±2.33c 625.00±2.13b 626.00±1.22a
Histidine 950.00±1.05g 951.00±1.53f 953.00±2.26e 955.00±2.136d 957.00±0.12c 958.00±2.71b 960.00±2.11a
Phenylalanine 1660.00±2.09g 1660.00±2.06f 1662.00±3.71e 1664.00±3.21d 1667.00±2.81c 1669.00±2.82b 1680.00±1.54a
Tyrosine 820.00±2.82g 820.00±2.13f 822.00±2.02e 823.00±2.33d 826.00±3.82c 828.00±2.01b 830.00±3.25a
Non-essential amino acids (mg/100g)
Arginine 5200.00±0.26e 5200.00±2.02e 5200.00±2.20e 5216.00±2.61d 5218.00±2.42c 5220.00±3.12b 5224.00±2.82a
Alanine 1420.00±1.32f 1420.00±1.32f 1422.00±2.42e 1425.00±2.28d 1428.00±2.07c 1430.00±3.02b 1434.00±2.04a
Glutamic 7000.00±2.42g 7005.00±2.42f 7008.00±2.42e 7010.00±2.20d 7016.00±2.13c 7020.00±2.21b 7028.00±2.32a
Aspartic acid 1620.00±1.76e 1620.00±2.19f 1620.00±3.37e 1621.00±2.02d 1623.00±2.42c 1626.00±3.42b 1630.00±2.13a
a, b, c, d= means within the same row bearing different superscript are significantly different (p<0.05)

Table 4: Effect of different boiling duration on anti-nutritive substances of Moringa oliefera seeds
Boiling duration in minutes
Parameters (mg/100g) 0 10 20 30 40 50 60
Tannin 241.00±1.56a 240.00±0.16b 216.0±0.26c 160.00±0.56d 140.00±1.56e 100.00±0.56f 68.00±2.23g
Phenolics 40.00±0.26a 34.00±1.26b 30.00±0.53c 20.00±0.16d 12.00±0.57e 8.00±0.26f 0.00
a b c d
Saponins 9.00±0.15 8.00±0.11 7.50±0.14 5.00±0.26 4.00±0.22e 3.50±0.23f 1.00±0.21g
Phytate 70.00±1.26a 65.00±0.11b 40.00±0.14c 25.00±0.82d 5.00±0.19e 0.00 0.00
a, b, c, d= means within the same row bearing different superscript are significantly different (p<0.05)

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