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© 2020 by the authors; licensee Asian Online Journal Publishing Group 
 

Agriculture and Food Sciences Research 
Vol. 7, No. 1, 28-37, 2020 

ISSN(E) 2411-6653/ ISSN(P) 2518-0193 
DOI: 10.20448/journal.512.2020.71.28.37 

© 2020 by the authors; licensee Asian Online Journal Publishing Group 

    
 

 
 
The Anti-Nutritional and Proximate Composition of Rain Tree (Samanea saman) 
Pod Samples 

 
Uzoukwu, A. E1   

Ubbaonu, C. N2   

Nwosu, J. N.3    

Ogueke, C. C.4   

Chukwu, M. N.5   

 

 
( Corresponding Author) 

 
1,2,3,4Department of Food Science and Technology, Federal University of Technology, Owerri, Imo State, Nigeria. 

 
5Department of Food Technology, Abia State Polytechnic, Aba, Abia State, Nigeria. 

 

 
Abstract 

The anti-nutritional and proximate composition of pods of Samanea saman were determined. 
About 600g of wholesome, cleaned and dried pods were divided into two portions: one portion 
was ground as whole pod meal while the other portion was deseeded and ground as seedless meal. 
The removed seeds were ground separately as a sample to obtain three samples namely: whole 
pod, pulp and seed samples. Triplicate data obtained were subjected to One-Way ANOVA using 
SPSS software of version 21. Mean values and Fisher’s least significant difference (LSD) were 
determined for the separation of the means at (p≤0.05). The whole pod had 1.49mg/100g oxalate, 
1.97mg/100g hydrogen cyanide, 0.71mg/100g tannins, 27.07mg/100g saponins, 0.51mg/100g 
flavonoids and 1.71mg/100g alkaloids, which were significantly(p≤0.05) different from that of 
pulp (1.89mg/100g oxalate, 1.51mg/100g hydrogen cyanide, 0.86mg/100g tannins, 
28.46mg/100g saponins, 1.00mg/100g flavonoids and 1.41mg/100g alkaloids) and seed samples 
(1.39mg/100g oxalate, 1.61mg100g hydrogen cyanide, 0.56mg/100g tannins, 26.51mg/100g 
saponins, 0.49mg/100g flavonoids and 1.86mg/100g alkaloids). The moisture contents were pulp 
(19.30%), whole pod (15.50%) and seed (9.20%). Ash content were whole pod (4.70%), pulp (2.90%) 
and seed (2.60%); Ether extract of whole pod, pulp and seed were 3.31%, 2.52% and 2.66% 
respectively. The crude proteins were whole pod (13.21%), pulp (10.98%) and seed (21.55%); crude 
fibre of the plant were whole pod (15.95%), pulp (6.77%) and seed (8.47%). The whole pod had 
47.33% carbohydrate which was significantly (p≤0.05) lower than the carbohydrate of pulp 
(57.53%) and seed (55.52%). The utilization of the pods of S. samana is a significant source of 
bioactive that if harnessed in the formulation of nutraceutical beverage could offer a whole lot of 
health benefit to the users. 

 
Keywords: Saponins, Tannin, Alkaloid, Oxalate, Flavonoid, Moisture, Proteins, Ash, Fibre, Carbohydrate. 

 
Citation | Uzoukwu, A. E; Ubbaonu, C. N; Nwosu, J. N.; Ogueke, C. 
C.; Chukwu, M. N. (2020). The Anti-Nutritional and Proximate 
Composition of Rain Tree (Samanea saman) Pod Samples. 
Agriculture and Food Sciences Research, 7(1): 28-37. 
History:  
Received: 2 January 2020 
Revised: 6 February 2020 
Accepted: 12 March 2020 
Published: 8 April 2020 
Licensed: This work is licensed under a Creative Commons 

Attribution 3.0 License  
Publisher:  Asian Online Journal Publishing Group 
 

Acknowledgement: All authors contributed to the conception and design of 
the study. 
Funding: This study received no specific financial support. 
Competing Interests: The authors declare that they have no conflict of 
interests. 
Transparency: The authors confirm that the manuscript is an honest, 
accurate, and transparent account of the study was reported; that no vital 
features of the study have been omitted; and that any discrepancies from the 
study as planned have been explained. 
Ethical: This study follows all ethical practices during writing.   

 

 

Contents 
1. Introduction ...................................................................................................................................................................................... 29 
2. Materials and Methods ................................................................................................................................................................... 29 
3. Results and Discussion ................................................................................................................................................................... 34 
4. Conclusion and Recommendation ................................................................................................................................................ 36 
References .............................................................................................................................................................................................. 36 
 

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Contribution of this paper to the literature 
This study has provided information for the utilization of the rain tree (Samanea saman) pod in food 
production. This study has also opened another area of research such as pod utilization in food 
formulation and animal feed development. 

 
1. Introduction 

The rain tree (Samanea saman Merr.) belongs to the family of leguminosae (pulse family) [1]. It is commonly 
called “Saman”, “Rain tree”, “monkey pod” and “Cow tarmarind” in English, while it is called “Acacia preta” and 
“Cenizaro” in Spain. The French call it “abre de Pluie” and the Philipines call it “Mimosa”. The name “Rain tree” 
was believed to have originated from the leaflets which are light-sensitive and close together on cloudy days or 
nights, allowing rain to fall through the leaves to the ground below [2]. It is native to Northern South America 
and has naturalized throughout the tropics. The plant is easily recognized by its characteristic umbrella-shaped 
canopy giving it a domed shape [3]. 

Samanea saman is a multipurpose tree and is the botanical name for rain tree, 5’O clock tree or monkey pod. S. 
saman is native to Northern South America but currently naturalized and distributed throughout the tropics. S. 
saman often planted in parks and pastures, vacant lots, churches and school grounds, along roadsides and planned 
landscape. S. saman is moderately fast growing with a growth rate of 0.75–1.5 m/year. Outdoors planting is done 
when the seedlings are 3–5 months old and 20–30 cm tall. It grows best in lowlands from sea level to 300 m high 
and rainfall of 600–3,000 mm [4, 5]. The pods of Samanea saman (Jack) Merr tree is straight, some-what fleshy, 
indehiscent, 15 to 20 centimeters long and 2 centimeter wide. When the pod ripens, the pulp is sweet and sugary 
with a flavor like licorice that is much relished by children. In South America, the sweet pulp is cooked like a 
tamarind. The seeds are oblong and reddish in color. Goats can eat the leaves and pods when feeds are scarce and 
the fruiting season of this tree coincides with the scarcity of good quality forages [6].  

The nutritional composition of Samanea saman pods and seeds as having crude protein values of 16.6% and 
31.6%, crude fibre values of 12.0% and 14.0%, ether extract values of 1.4% and 4.3%, ash values of 3.5% and 4.3%, 
carbohydrate values of 18.5% and 20.3% respectively. The ripe fallen pod is commonly eaten by children and 
animals [7] while the sweet, sticky pulp is used in the production of fruit drink similar to tamarindo in Latin 
America [2, 8].  

The pods are edible and eagerly eaten by human and livestock (cattle, goat) both domesticated and wildlife. 
The seed of S. saman is already a good source of protein and energy. It contains 13.57% protein, 89.25% dry matter, 
2.98%ether extract (EE), 2.19% crude fibre, 0.23% ash, and 6.44% nitrogen extract. Apart from its nutrient content 
which is almost comparable to corn, the pod is easily available making it a cheap source of feed for chickens Cruz 
[9]. Staples and Elevitch [2] reported that rain tree pods are nutritious with 12–18% protein, 40% digestibility 
and because of the sweet pulp are eagerly eaten by cattle, hogs, horses and goats. According to Barcelo and Barcelo 
[6] fresh Samanea saman (Jack) Merr pods had dry matter content of 84.82%, crude protein of 9.45%, crude fiber of 
8.34%, crude fat of 8.82% and ash of 5.12%. The Samanea saman (Jack) Merrpod is rich in carbo-hydrates as 
reflected in its nitrogen-free extract of 53.09%. Samanea saman (Jack) Merr pods are relished by livestock due to its 
sweet taste. The seeds are also surrounded by sticky, gummy substance. 

The whole pods are used as feed and nutrient supplements for mammals such as squirrels, cows, cattle and 
goats. Okonkwo [10] reported that its seed has been used in the production of a local condiment called Anyu, 
However, the few works done on this pod were directed towards determining their anti-microbial and 
phytochemical potentials, and the use of their seeds in the production of “Anyu” a local condiment [10-12]. 
Available literature shows there is little or no research work on the use of these pods in their utilization in other 
commercial food products such as bread, yet these pods are wasting in the gardens and road-sides where the trees 
are planted for ornamental purposes. Even in the campus of Federal University of Technology, Owerri (FUTO), 
the trees fruit heavily and animals (cows or others) at their will pick up the heavily littering pods within the 
fruiting season [6]. 

Anti-nutrients are natural or synthetic compounds that interfere with the absorption of nutrients [13]. Anti-
nutritional factors are present in different food substances in varying amounts, depending on the kind of food, mode 
of its propagation, chemicals used in growing the crop as well as those chemicals used in storage and preservation 
of the food substances [14, 15]. Some oil seeds and legumes used in the production of foods contain toxic 
substances and anti-nutritional factors; for example, tannin in castor oil bean, trypsin inhibitors in soybean, 
oxalates in locust beans, phytic acid in African oil bean, phytate in melon seeds [16, 17]. Despite the protein 
quality in leguminous seeds, it does not however reach the same level as in animal products. This is due to 
unbalanced amino acids, presence of anti-nutritional factors and low digestibility of protein. Processing methods, 
such as soaking, cooking or fermentation can improve the quality of legume protein [18]. During fermentation, 
micro flora may produce proteolytic enzymes which may be responsible for the increase in protein digestibility. 
Also, the elimination of phytic acid contributes to the improvement in protein digestibility of fermented products 
[19]. 

The main objective of this research is to investigate the anti-nutritional and proximate composition of rain tree 
pod. The specific objectives are to analyze the anti-nutrients and proximate composition of Samanea saman whole 
pods, pulp and seeds. The notable sweetness of the rain tree pulp when applied in food is hoped to provide an 
alternative source of sugar for food production, with a resultant reduction in the cost of production and 
consequently the price of food in the market. Besides, it’s distinct but attractive flavour may introduce a variety in 
food products. A good result from this study will certainly introduce the underutilized pods to the food industry 
and add economic value to it. 
 

2. Materials and Methods 
2.1. Collection of Materials 

Some ripe, fallen wholesome rain tree (Samanea saman) pods were picked from the tree sites around the School 
of Agriculture and Agricultural Technology (SAAT) of the Federal University of Technology, Owerri (FUTO), 



Agriculture and Food Sciences Research, 2020, 7(1): 28-37 

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Imo State, Nigeria. Some of the chemicals used for the analyses were obtained from the Department of Food 
Science and Technology laboratory, and School of Agriculture and Agricultural Technology laboratory in FUTO. 
Others were purchased from the Finlab Store Owerri, Imo State, Nigeria. Some of the analyses were conducted in 
the laboratories of the Department of Food Science and Technology and School of Agriculture and Agricultural 
Technology, FUTO, while others were carried out in the Department of Zoology laboratory of University of Jos 
and International Institute for Tropical Agriculture (IITA), Ibadan, Oyo State, Nigeria.  
 

2.2. Methods 
2.2.1. Preparation of Samples  

The pods were picked at various times from the tree site, sorted, washed and sun dried for about five days. Six 
hundred grams (600g) of wholesome, cleaned and dried pods were divided into two portions; one portion was 
ground as whole pod meal (sample 1) shown in Figure 1 and Figure 2, while the other portion was deseeded and 
ground as seedless meal (sample 2) as shown in Figure 2. Then the removed seeds Figure 3  were collected and 
ground separately as a sample (Sample 3) to obtain three samples for the desired raw material analyses, namely 
whole pod, pulp and seed samples [20]. 

 

 
Figure-1. A Rain Tree (Samanea saman) Plant in FUTO. 

 

 
Figure-2. Rain tree (Samanea saman) Pods. 

 



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Figure-3. Rain tree (Samanea saman) Seeds. 

 

2.2.2. Determination of Anti-Nutrients in the Pods of Rain Tree 
2.2.2.1. Saponin Content 

The method described by Okwu [21] was used for the analyses of the anti-nutrients. Twenty-five grams (25g) 
of the sample was placed in a two hundred and fifty milliliters (250ml) flask. One hundred milliliters (100ml) of 
20% (V/V) aqueous solution of ethanol was poured into the flask; then the flask with the content was placed in a 
hot water bath at 55OC for 4 hours with occasional stirring. The mixture was filtered and the residue re-extracted 
with a fresh 100ml of 20% (V/V) ethanol solution as before. Both extracts were combined and the volume reduced 
to about 50ml in a water bath set at 90OC. The mixture was allowed to cool, extracted with 20ml of diethyl ether in 
a 250ml separating funnel. The ether layer was discarded and 50ml of n-butanol was poured into the lower layer. 
The mixture was shaken and the two layers allowed separating. The organic layer was washed twice with 10ml of 
5%(V/V) sodium chloride solution in a separating funnel. This was poured into a pre-weighed beaker and 
evaporated to dryness on a boiling water bath. The beaker was cooled and weighed again [22]. The percentage 
saponin content was determined using Equation 1. 

%𝑆𝑎𝑝𝑜𝑛𝑖𝑛 =
𝑊3 − 𝑊2

𝑊1
×

100

1
                                            (1)              

Where 
W3 = Weight of beaker and residue after evaporation to dryness. 
W2 = Weight of beaker alone. 
W1 = Weight of sample. 
 

2.2.2.2. Flavonoid Content 
Two grams (2g) of the ground sample was placed in a 250ml Erlenmeyer flask and 100ml of 80% (V/V) 

methanol solution was added. The mixture was stirred with a magnetic stirrer for 3 hours and filtered with 
Whatman filter paper (No. 42). The residue was extracted with a fresh 100ml of 80% (V/V) methanol as before and 
the mixture filtered again. Both extracts were combined into a pre-weighed beaker and evaporated to dryness on a 
boiling water bath. This was cooled and weighed again [23]. The percentage flavonoid content was determined 
using Equation 2.  

%𝐹𝑙𝑎𝑣𝑜𝑛𝑜𝑖𝑑 =
𝑊3 − 𝑊2

𝑊1
×

100

1
                                    (2) 

Where: 
W3 = Weight of beaker and flavonoid. 
W2 = Weight of beaker. 
W1 = Weight of sample. 
 

2.2.2.3. Determination of Alkaloid Content 
Two grams (2g) of the ground sample was placed in a 250ml flask and 100ml of 20% (V/V) acetic acid in 

ethanol was added to the flask. The flask was covered and allowed to stand for 4 hours while stirring with a 
magnetic stirrer. The mixture was filtered and the residue re-extracted with a fresh 100ml of 20% (V/V) ethanolic 
acetic acid solution. Both extracts were combined in a beaker and concentrated on a water bath to about one 
quarter of the original volume. The concentrate was cooled and concentrated ammonium hydroxide added in drops 
until complete precipitation of the alkaloids had occurred. The resulting mixture was allowed to stand overnight. 
Thereafter, it was filtered using a pre-weighed filter paper. The filter paper with the content was dried in an oven 
set at 70OC for 6 hours and cooled to ambient temperature in a desiccator. The weight of the filter paper and its 
contents were taken [24, 25]. The percentage alkaloid content was determined using Equation 3. 

%𝐴𝑙𝑘𝑎𝑙𝑜𝑖𝑑𝑠 =
𝑊3 − 𝑊2

𝑊1
×

100

1
                                       (3)  

Where 
W3 = Weight of filter and alkaloids after drying. 
W2 = Weight of filter paper alone. 
W1 = Weight of sample. 



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2.2.2.4. Determination of Tannin Content  
About 0.5grams portion of the ground sample was placed in a 100ml flask and fifty milliliters (50ml) of distilled 

water was added and the mixture was stirred for about one hour with a magnetic stirrer. The sample was filtered 
with a moist filter paper into a fifty milliliters (50ml) volumetric flask and made up to the mark with distilled water. 
Five milliliters (5ml) of the filtrate was pipette into a tube and mixed with three milliliters (3ml) of 0.1ml of ferric-
chloride in 0.1N hydrochloric acid and 0.008M potassium ferrocyanide solution. The absorbance was measured in a 
spectrophotometer set at 120nm wavelength within ten (10) minutes of mixing. A blank sample was prepared and 
the colour was developed and read at the same wavelength. A standard was prepared using tannic acid to get 
100ppm and measured [26]. The tannin content was calculated using the Equation 4: 

𝑀𝑔 𝑚𝑙 𝑇𝑎𝑛𝑛𝑖𝑛 =⁄  𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑋
𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑

1
               (4) 

 

2.2.2.5. Determination of Cyanogenic Glycoside Content 
One gram (1g) of the ground sample was placed in a round bottom flask and 200ml of de-ionized distilled water 

was added and the flask allowed standing for 2 hours. A distillation unit was set up and about 150mls of the 
reaction mixture (sample and de-ionized distilled water) was distilled into a 250ml receiver flask containing 20ml of 
2.5% (W/V) sodium hydroxide solution. One hundred milliliters (100ml) of the mixture was measured into a 250ml 
flask and 8ml of 6M ammonium hydroxide solution was added followed by 2ml of 5% potassium iodide solution. 
The content of the flask was mixed and titrated with 0.02N silver nitrate solution till there was no further turbidity 
[27]. The percentage cyanogenic glycoside content was determined using Equation 5.  

%𝐶𝑦𝑎𝑛𝑜𝑔𝑒𝑛𝑖𝑐 𝑔𝑙𝑦𝑐𝑜𝑠𝑖𝑑𝑒 = 𝑇 × 108 ×
𝑁

1000
×

𝑉𝑒

𝑉𝑎
×

100

𝑊
       (5) 

Where 
T = Titre value. 
108 = Equivalent weight of silver nitrate. 
N = Normality of silver nitrate solution. 
Ve= Extract volume. 
Va= Aliquot volume titrated. 
W= Weight of sample in grams. 
1000 = Scaling factor to obtain silver nitrate content of 1.0ml of silver nitrate solution. 
 

2.2.2.6. Determination of Oxalate Content 
Five grams (5g) of the ground sample was placed in a 100ml flask and 20ml of 0.3N hydrochloric acid solution 

was poured into the flask. It was stirred with a magnetic stirrer at 50OC for I hour. The mixture was filtered with 
filter paper (No. 42) and the extract stored and the process was repeated. Both extracts were combined and made 
up to a volume of 100mlwith distilled water. Twenty milliliters (20ml) of the filtrate was poured into a 100ml 
beaker and three drops of phenolphthalein indicator was added. Then, 5N ammonium hydroxide solution was 
added in drops until the reaction mixture was alkaline (pinkish). Glacial ethanoic acid was added in drops until the 
pink coloration disappeared and a few more drops were added to make the mixture acidic. Next, 5ml of 5% calcium 
chloride solution was added to the solution and the mixture was allowed to stand for 3 hours. It was centrifuged at 
300r.p.m for 15 minutes and then the residue washed 3 times with hot water using a centrifugation technique. The 
residue was dissolved in 3ml of 3N tetraoxosulphate (VI) acid solution. The resulting solution was titrated with 
freshly prepared 0.01N potassium permanganate solution until permanent pink coloration that lasted for about 30 
seconds was obtained [28]. A blank titration was also conducted. The percentage oxalate content was determined 
using Equation 6. 

%𝑂𝑥𝑎𝑙𝑎𝑡𝑒 =
𝑋

1
×

1

1000
×

100

1
𝑎 ×

100

𝑆
𝑏                                (6) 

Where 
X = Weight of oxalate obtained by multiplying molar mass and molarity of oxalate. 
1000 = Reference volume for molar concentration. 
100a = Total volume of extract. 
100b = Scaling factor to convert to percentage. 
S = Weight of sample taken for analysis. 
 

2.2.3. Proximate Composition of the Samples of Pods of Rain Tree 
2.2.3.1. Determination of Moisture Content 

The AOAC [29] method was used. Two gramme-portions of each of the freshly ground samples (1, 2 and 3) 
were weighed into previously weighed dry crucibles. The crucibles with samples were dried in an oven at 105oC, 
cooled in desiccators for ten minutes, reweighed and returned into the oven until a constant weight was attained 
[26]. The moisture content was calculated with the following Equation 7: 

% Moisture content = 
𝑊1−𝑊2

𝑊1
×

100

1
                            (7) 

Where: 
W1 = the weight of sample before drying. 
W2 = the weight of sample after drying. 
Sample 1= Whole pod, Sample 2= pod without seed (pulp), Sample 3= seed. 
 
 
 



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2.2.3.2 Determination of Ash Content 
The Ash content was determined following the AOAC [29] method. Two grammes of the samples were 

weighed in triplicate into previously weighed silica crucibles. The samples inside the crucibles were charred on a 
heater inside a fume cupboard to drive off most of the smoke. The crucibles with the contents were transferred into 
a muffle furnace and heated for about four hours at 550OC. They were cooled in a desiccator and weighed. The 
heating was repeated until the samples turned greyish white and attained constant weight. The ash content was 
then calculated as shown in Equation 8: 

%Ash  =    
𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑎𝑠ℎ

𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑡𝑎𝑘𝑒𝑛
×

100

1
   (8) 

 

2.2.3.3. Determination of Crude Lipid (Ether - Extract) 
The total lipid content of the samples was determined using the AOAC [29] Soxhlet fat extraction method.  

Five grammes (5g) of the sample were weighed into a pre-weighed fat free extraction thimble which was plugged 
tightly with cotton wool. The thimble was placed in the Soxhlet extractor fitted up with reflux condenser, all 
connected to a boiling flask containing 200ml of petroleum ether (Boiling point 60oC) on a heating mantle. As the 
flask and petroleum ether were heated, the solvent evaporated and condensed into the thimble extracting oil from 
the sample and refluxed into the boiling flask with the extracted oil. This was done for 4 hours. At the end of 
extraction, the solvent (petroleum ether) was evaporated by heating at 70oC on a hot plate leaving the lipid extract 
in the flask. The flask with its contents were placed in an oven and dried at 110oC for one hour, cooled in a 
desiccator and re-weighed [26].  The percentage lipid was calculated as shown in Equation 9: 

%Ether extract 
𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑜𝑖𝑙

𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒
×

100

1
   (9) 

 
2.2.3.4. Determination of Crude Protein 

The crude protein contents of the samples were determined using the micro-Kjeldahl apparatus as described by 
AOAC [29]. Two grammes of each of the samples was placed in a Kjeldahl flask and 30ml concentrated sulphuric 
acid (H2SO4) added followed by the addition of 10g potassium sulphate and 1g copper sulphate. The mixture was 
gently heated for few minutes until frothing ceased; the heat was increased and the sample allowed digesting for 
three hours. The digest was allowed to cool, diluted with distilled water (washing the digestion flask) up to 100ml. 
Ten milliliters(10ml) of the dilute digest was pipetted into a distillation flask and 10ml of 40% (w/v) sodium 
hydroxide added. The mixture was distilled and the liberated ammonia collected in 10ml of 2% boric acid 
containing indicator. This was titrated with 0.01N hydrochloric acid to grey coloured end point. A blank was also 
prepared without a sample and treated as above [26]. The amount of crude protein was then calculated by 
multiplying percentage nitrogen in the digest by the conversion factor (6.25) as shown in Equation 10.  

 

%𝑁 =
(𝑎 − 𝑏) × 0.01 × 14 × 𝑣

𝑊 × 𝐶
×

100

1
        (10) 

Where 
a = the titre value of the digested sample. 
b = titre value of the blank sample. 
V = volume after dilution. 
W = weight of dried sample (mg). 
C = Aliquot of sample used. 
14 = Atomic weight of Nitrogen. 
Crude protein = 6.25 x %N. 
 

2.2.3.5. Determination of Crude Fibre 
AOAC [29] method was employed. Two grams of the ground samples were weighed in duplicate into a 600ml, 

long Pyrex beaker and 200ml of 1.25% H2SO4 solution was added. The beaker was covered with a watch glass and 
the content gently boiled on a hot plate for 30 minutes. The acid was removed by filtering through a muslin cloth 
over a Buckner funnel and the sample washed three times with 50ml of boiling water to free it of acid, before 
putting it back to the beaker. Then, 200ml of 1.25% NaOH solution was added to the residue in the beaker, which 
was covered with a watch glass and gently boiled on a hot plate for 30 minutes and then filtered. The residue was 
washed into a weighed No. 2 sintered glass crucible with 50ml of boiling water and later washed twice with 30ml 
portions of petroleum spirit. The crucible was dried in the oven at 80OC to a constant weight and then ignited in a 
muffle furnace at 600OC until a light gray coloured ash was obtained. The crucible and content were cooled to 
ambient temperature in a desiccator and then weighed [26]. The crude fibre content was calculated as shown in 
Equation 11: 

% crude fibre =
𝑙𝑜𝑠𝑠 𝑖𝑛 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑛 𝑖𝑔𝑛𝑖𝑡𝑖𝑜𝑛

𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒
×

100

1
                           (11) 

 

2.2.3.6. Determination of Carbohydrate Content 
The carbohydrate content was obtained by difference [26] as shown in Equation 12.  
 
%Carbohydrate = (100% - %Moisture, %Crude protein - %Fat-%Ash-%Crude fibre)     (12) 

 
2.2.3.7. Statistical Analysis 

Triplicate data obtained were subjected to statistical analysis using SPSS software of version 21.  Mean values 
were determined and One-Way ANOVA was done as well as Fisher’s Least Significant Difference [30] was used to 
determine for the separation of the means at (p≤0.05). 
 



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3. Results and Discussion 
3.1. Anti-Nutritional Composition (Mg/100g) of Rain Tree (Samanea saman) Pod Samples 

Oxalate, hydrogen cyanide, tannins, saponins, flavonoids and alkaloids were identified in all samples of Samanea 
saman pod Table 1.  

Oxalic acid as an anti-nutrient interfere with mineral availability particularly calcium. It binds with calcium 
and forms insoluble calcium oxalate which cannot be absorbed in the body. This may lead to death due to 
hypercalcimia in the renal tubules Murray, et al. [31]; Ifemeje [32]. Ladeji, et al. [33] reported that oxalates cause 
irritation and swelling in the month and throat. The oxalate of the pulp sample was 1.89mg/100g which is 
significantly (p<0.05) higher than the oxalate (1.49mg/100g) of whole pod and 1.39mg/100g oxalate of seed 
sample Table 1. This quantity of oxalate is within safe limit because 2% or more soluble oxalate can lead to 
toxicosis. They are therefore considered harmless when present in small amounts as shown in Table 1. The amount 
of oxalate in Samanea saman whole pod, pulp and seed samples is thus not harmful, more so, when cooking has been 
reported [34] to affect a significant reduction in total oxalate contents of plants. An unacceptably high value of 
27.34 mg/100 g oil was reported for tropical almond [35]. 

Cyanogenic glycosides are hydrolysed by β-glucosidase producing sugars and a cyanohydrin which 
spontaneously decompose to cyanohydric acid (HCN) and a ketone or aldehyde [36]. HCN is extremely toxic to a 
wide spectrum of organisms due to its ability of linking with metals (Fe2+, Mn2+ and Cu2+) that are functional 
groups of many enzymes, inhibiting processes like reduction of oxygen in the cytochrome respiratory chain, 
electron transport in the photosynthesis, and the activity of enzyme like catalase, oxidase [32]. Hydrogen cyanide 
is toxic when ingested by monogastric animals in large quantity [37]. The whole pod had HCN of 1.97mg/100g 
which is significantly (p<0.05) higher than the HCN (1.51mg/100g) of pulp and 1.61mg/100g of seed samples 
Table 1. The hydrogen cyanide contents of these samples were low when compared to the report (4.31-4.77mg/kg) 
of Ogueke, et al. [38] and maximum limit (10 mg/kg) recommended for garri [39]. Reduction in HCN might be 
due to the processing methods employed [40]. High level of hydrogen cyanide has been implicated for cerebral 
damage and lethargy in man and animals [34, 41]. 

Tannins are plant polyphenols, which have ability to form complexes with metal ions and with macro-
molecules such as proteins and polysaccharides [42]. Dietary tannins are said to reduce feed efficiency and weight 
gain in chicks [37]. Tannins are polyphenols and have been reported to exhibit anti-microbial actions. The 
presence of high level of tannins could confer on the users chemoprotective benefits [43]. The pulp had tannins of 
0.86mg/100g which is significantly (p<0.05) higher than the tannins (0.71mg/100g) of whole pod and 
0.56mg/100g of seed samples Table 1. Tannins may form a less digestible complex with dietary proteins. 
 

Table-1. Mean Values (mg/100g) of Anti-Nutrient Composition of Rain Tree (Samanea saman) Pod samples 

 SAMPLES (mg/100g) 

Anti-Nutrients Whole pod Pulp Seed LSD 

Oxalate 1.49±0.01b 1.89±0.01a 1.39±0.01c 0.02 
Hydrogen Cyanide 1.97±0.01a 1.51±0.01c 1.61±0.01b 0.02 

Tannins 0.71±0.01b 0.86±0.01a 0.56±0.01c 0.02 
Saponins 27.07±0.01b 28.46±0.01a 26.51±0.01c 0.02 

Flavonoids 0.51±0.01b 1.00±0.01a 0.49±0.01c 0.02 
Alkaloids 1.71±0.01c 1.41±0.01a 1.86±0.01b 0.02 

        Note:  Means with different superscripts on the same row are significantly (p< 0.05) different. 
 
These tannins-protein complexes are astringent and adversely affect feed intake. The pod contains 

0.86mg/100g of tannin due to this result taking the pod may not lower the availability of protein in the body or 
clotting with red blood cell as cause by excess tannin in human body. This suggests that this pod may be safe for 
consumption [44]. 

Saponins are glycosides, which include steroid saponins and triterpenoid saponins [42]. High levels of 
saponins in feed affect feed intake and growth rate in poultry. Reduction in feed intake has been ascribed to the 
bitter taste of saponins [37] and due to the irritating taste. Excess saponins causes hypocholestrolaemia because it 
binds cholesterol making it unavailable for absorption [45]. Saponin-protein complex formation can reduce protein 
digestibility [37]. Saponins were predominant in all the Samanea pod samples. The highest level of saponin was 
found in Samanea saman pulp (28.46mg/100g), followed by the whole pod (27.07mg/100g) and the seed 
(26.51mg/100g). High levels of saponins have haemolytic effect on red blood cells and also reduce growth rate in 
animals. Saponins also possess some beneficial properties such as cholesterol lowering, anti-tumor and as 
antioxidant [33]. The concentration of the value of saponins from the analysis was 28.46mg/100g which is within 
the WHO permissible limit of (48.50mg/100 g) as recommended in this study. Saponins possess a carbohydrate 
moiety attached to a triterpenoid or a steroidal aglycone [46]. Saponins reduce the uptake of glucose and 
cholesterol at the gut through intra-lumenal physico-chemical interactions. This could confer a chemo-protection 
against heart diseases to users [32]. 

Flavonoids, a group of natural substances with variable phenolic structures, are found in fruits, vegetables, 
grains, bark, roots, stems, flowers, tea and wine. The flavonoids are categorized in different classes as alkaloids, 
terpenoids and phenolics. Flavonoids carry out a number of protective functions in the human body. Many 
flavonoids have evolved as bioactive compounds that interfere with nucleic acid or proteins and show antimicrobial 
or insecticidal and pharmacological properties [47].The highest level of flavonoids was found in Samanea saman 
pulp (1.00mg/100g), followed by the whole pod (0.51mg/100g) and the seed (0.49mg/100g) as shown in Table 1. 
Flavonoids possess anti-inflammatory, anti-oxidant, anti-allergic, hepato-protective, anti-thrombic, anti-viral and 
anti-carcinogenic activities. These properties may explain the use of Samanea saman for prevention and treatment 
of diseases [47]. 

Alkaloids are beneficial chemicals to plants with predator and parasite repelling effects. However, they inhibit 
certain mammalian enzymic activities such as those of phosphor-diesterase, prolonging the action of cyclic AMP. 
They also affect glucagons and thyroid stimulating hormones, while some forms have been reported to be 



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carcinogenic [48]. Alkaloids were relatively lower in the Samanea saman pulp (1.41mg/100g) than the seed 
(1.86mg/100g) and whole pod (1.71mg/100g) as shown in Table 1. It is noteworthy that at the concentration of 
alkaloids in edible plants, they are usually non-toxic because steaming or boiling reduces their levels in plant 
extracts [32]. 
 

3.2. Proximate Composition of Rain Tree (Samanea saman) Pod Samples 
The fresh mature and ripe (brown) rain tree whole pod had a moisture content of 15.50% and understandably 

its pulp and seed had moisture contents of 19.30% and 9.20% respectively Table 2. The values were significantly 
(p≤0.05) different from each other. Esuoso [8] found that Samanea saman husks, whole pods and seeds had 
moisture contents of 4.11%, 58.42% and 6.19%respectively. The slight variation in moisture content could be 
attributed to the drying conditions which the samples were subjected to before the analysis. Factors such as 
environmental conditions could also affect the moisture content of food materials. Barcelo and Barcelo [6] showed 
that the Samanea saman pod had a moisture content of 15.18%. The moisture content of 15.50% for the whole pod 
obtained in this study is in line with the values 15.30% and 15.80% reported by Barcelo and Barcelo [6]. The 
moisture content of foods determines the water activity which influences the rates of chemical, microbial and 
enzymatic reactions in foods. By lowering the water activity through drying, microbial deterioration of foods can 
be delayed, reduced or eliminated. 

The whole pod had higher ash content (4.70%) than the pulp (2.90%) or the seed (2.60%), indicating that it has 
a higher concentration of minerals than pulp and seed. This is in line with Esuoso [8] who reported ash contents of 
4.16% and 2.10% for Samanea saman whole pod and seed respectively. Barcelo and Barcelo [6] reported a relatively 
higher ash content of 5.12% for Samanea saman whole pod. Though, there were slight variations in the ash contents 
of the samples, the values fall within a close range of 2.4% to 5.12%. Ash in food constitutes the residue remaining 
after all the moisture has been removed as well as the organic materials burnt away by igniting at temperature of 
about 500oC [49]. Thus, ash content represents the total mineral content in foods. 
 

Table-2. Mean values of the proximate composition of rain tree (samanea saman) pod samples. 

  Samples 

No. Proximate 
Composition (%) 

Whole pod Pulp Seed LSD 

1 Moisture 15.50±0.01b 19.30±0.01a 9.20±0.1c 0.02 
2 Ash 4.70±0.01a 2.90±0.01b 2.6±0.01c 0.02 
3 Ether Extract 3.31±0.01a 2.52±0.01c 2.66±0.01b 0.02 
4 Crude Protein 13.21±0.01b 10.98±0.01c 21.55±0.01 0.02 
5 Crude Fibre 15.95±0.01a 6.77±0.11c 8.47±0.01b 0.13 
6 Carbohydrate 47.33±0.01c 57.53±0.01a 55.52±0.01b 0.02 

Note:  Means with different superscripts on the same row are significantly (p< 0.05) different. 

 
The lipid content (Ether extract) results in Table 2 shows that Samanea saman pod had the highest lipid 

content(3.31%) among the samples with Samanea saman seed having 2.66% and the Samanea saman pulp having the 
lowest lipid content (2.52%).The lipid content of all the samples were significantly (p< 0.05) different from each 
other. Similarly, Esuoso [8] reported a lower lipid value of 0.82% for the whole pod and 1.20% lipid content for the 
seed. The variations observed in the lipid contents could be as a result of agronomical factors and analytical 
techniques. With ether extract (fat) values of the samples ranging from 2.52% (in pulp) to 3.31% (in whole pod), 
certainly the pod is not a good oil source, thus its seed could not be classified as an oil seed. 

Among the three areas of Samanea saman pod studied, the seed had the highest protein value (21.55%). The 
whole pod had protein value of 13.21% while its pulp had a value of 10.98%. The differences between these values 
were significant (p<0.05). The protein value of 13.21% obtained in this study for the whole pod fell within the 
range of 10-18% reported by Dihigo, et al. [50] and Staples and Elevitch [2]. Semae, et al. [51] reported that the 
pods of rain tree to be highly digestible, having protein (15.31–18.00%) content, is low in cost and is non-toxic [52, 
53]. Esuoso [8] reported a whole pod protein value of 28.48% and that was the highest value observed in 
accessible literature for the pod. There was no specific report on the pulp with regards to proximate composition, 
but reports on the protein content of seed had values ranging from 22-31.60% [8, 50]. Notwithstanding, the 
variations observed in the various reports, Samanea saman pods and seeds exhibited high protein contents which 
could be explored in animal feed as nutrient source to improve animal diet and possibly for humans. 

The Samanea saman whole pod had a mean crude fibre content of 15.95% as compared to values of 8.47% and 
6.77% for the seed and pulp respectively. The crude fibre content of the three samples were significantly (p<0.05) 
different. The crude fibre value of 15.95% obtained in this study for the whole pod is comparably much lower than a 
value of 33.4% reported by Esuoso [8] for the same material. Also, a crude fibre value of 8.47% seed obtained in 
this study is comparably lower than the value 16.15% reported by Esuoso [8] respectively for the same material. 
Though agronomical factors may have contributed to the variations in the results obtained, different analytical 
conditions may have also led to the variation. Crude fibres play important roles in digestion processes and the 
lowering of gastric cholesterol [21]. 

Table 2 showed that Samanea saman pulp had the highest carbohydrate content (57.53%) as compared to the 
seed with a value of 55.52% and the whole pod with the least carbohydrate content of 47.33%. There were 
significant (p<0.05) differences among the carbohydrate contents of the three samples. Since the carbohydrate 
content was obtained by difference, the lower carbohydrate content observed in the whole pod could be a reflection 
of the higher crude fibre content of the whole pod which was much higher than that of the Samanea saman pulp and 
seed. The result obtained in this research showed some similarities with those previously reported by other 
researchers, where the seed had higher carbohydrate than the pod. Esuoso [8] also reported lower carbohydrate 
values of 26.73% and 34.89% for the Samanea pod and seed, respectively. The relatively high carbohydrate content 
obtained in this research suggests that the various parts of Samanea saman pods could serve as energy source when 
included in animal feed. 



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4. Conclusion and Recommendation 
4.1. Conclusion 

The levels of hydrogen cyanide, oxalate, tannins and flavonoid were relatively low in all the samples of 
Samanea saman pods. The results show that all the anti-nutrients were significantly (p<0.05) different in all the 
samples. The relative low levels of these anti-nutrients confirm the safety of the fruit for industrial utilization, 
human and animal consumption. It was observed that saponins were the predominant anti-nutrient identified in all 
the pod samples ranging from 26.51mg/100g (seed) to 28.46mg/100g (pulp) while the rest were less than 
2.0mg/100g. The result of the anti-nutritional composition of Samanea saman pods is evidence that utilization of 
Samanea saman might not have any negative impact in the body. Though saponin was the most abundant anti-
nutrient in all the Samanea saman pod samples, the highest level (26.51mg/100g) identified in the pulp is still 
within the safe level of less than 146mg/100g. 

The result of proximate composition of pod of rain tree showed that pulp had higher moisture content and 
carbohydrate content than whole pod and seed samples. It was observed that the whole pod had higher ash, ether 
extract and crude fibre than the pulp and seed samples whereas the seed was richer in protein content than the 
whole pod and pulp samples.  The relative high protein (10.98% to 21.55%) and carbohydrate (47.33% to 57.53%) 
contents of the rain tree (Samanea saman) pod samples suggest that they can be used as nutrient supplement in 
animal feeds and the seeds can be utilized as a rich protein meal in human diets. 
 

4.2. Contributions to Knowledge 
Within the limits of this research work, the study has, no doubt, contributed immensely to knowledge, which is 

as follows: 

• This study has provided information for the utilization of the rain tree (Samanea saman) pod in food 
production.  

• This study has also opened another area of research such as pod utilization in food flavor and animal feed 
development. 

• The study has revealed the nutrient (protein, carbohydrate, lipid, ash, fiber and moisture) potential of these 
pods which are presently littering as waste. 

• This study has also opened another area of research such as pod utilization in food formulation and animal 
feed development. 

 
4.3. Recommendations 

This study leads to the need of further detailed search on the immense potentials of rain tree. The results of the 
present study are promising thus indicating the utilization of the pods of S. saman as a significant source of 
bioactive that if harnessed in the formulation of nutraceutical beverage could offer a whole lot of health benefit to 
the users. Investors should be encouraged to invest in machines that can facilitate the removal of seeds from the 
pods. 
 

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