Article Type : Research Article
Authors : Rajbongshi N, Khalek A, Rafia R, Miah S, Amin A, Ahmed S, Islam MS and Hosen A
Keywords : Moringa oleifera; Proximate analysis; Preservation
Moringa
oleifera leaves have
nutritional and medicinal values that helps to prevent disease. The objective
of present research was to investigate the nutritional value and
microbiological evaluation of Moringa oleifera leaves powder during storage
periods. In these research leaves were collected from Dhaka, savar, dhamrai,
nabinagar, kaliakoir respectively. Moringa
oleifera leaves were processed by using 3 different methods like as solar
dried, mechanical drier and oven drier. Nutritional values (proximate analysis)
including moisture, protein, ash, fat, carbohydrates and energy were measured
by utilizing distinct methods and devices. This study found highest moisture
content in solar drier at 4th month of storage Moringa oleifera powder. Moisture content of raw Moringa oleifera is 81.4% where
gradually increased the moisture content during the storage periods. Three
different methods including solar drier, mechanical drier and oven drier were
used for measuring nutritional values. The result showed that in solar drier,
moisture content was measured increasing during 4 month storage 3.93%, 4.06%,
4.11% and 4.52% where mechanical drier were 7.75%, 7.80%, 7.86%, 7.96% and oven
drier 6.15%, 6.28%, 6.75% and 6.82%. Other proximate analysis also measured
both three methods. Beside proximate analysis microbiological analysis also
conducted in the storage periods. Highest number of coliform bacteria and
fungus were identified in the last month of storage periods. Among three
methods oven drier gives best results during preservation periods. Moringa oleifera leave could be used as
antimicrobial medicine against commercial antibiotics and also cure different
diseases such as diarrhea, high blood pressure, food poisoning etc.
Moringa oleifera is a perennial tree that belongs to the Moringaceae
family and is yet underused. Drumstick, sahjan, and sohanjana are all names for
the same plant. All plant components have a wide spectrum of functional and
nutraceutical qualities, making this plant a versatile biomaterial for food and
other applications [1]. The leaves, blooms, and fruits of this plant are utilized
as a major food source to battle protein energy deficiency, which affects the
majority of the world's developing and underdeveloped countries. Because of the
presence of several types of antioxidant chemicals, this plant's leaves are
very good origin of nutraceuticals and functions [2]. Flavonoids from plants
are significant in the diet, and a high flavonoid consumption has been
associated to a lower risk of cardiovascular disease, osteoporosis, and are
other age related degenerative complication [3-5]. The World Health
Organization (WHO) has been studying M. oleifera's usage as a low-cost
supplement enhancer in the world's poorest areas for decades (WHO Readers
Forum, 1999). According to the United Nations Food and Agriculture
Organization, one out of every twelve people in the world is malnourished, with
160 million children under the age of five falling into this category (United
Nations Food and Agriculture Statistics, 2008). Herbal remedies are still
widely utilized in many parts of the globe, particularly in places where modern
treatments are unavailable [6]. Furthermore, scientific study to establish the
biological activity of medicinal plants is needed in utmost African nations
where verdant treatments are yet widely depended on due to the expense of chemotherapy
treatments. The findings of this research might lead to the validation of
historically used and medicinally important plants, allowing them to be fully
utilized [7]. Moringa is a fast-growing plant that is commonly accessible in
the tropics and subtropics and has a wide range of industrial and therapeutic
applications. One of the most extensively farmed species is Moringa oleifera, an essential medical
plant. Anti-tumour, antioxidant, anti-inflammatory/diuretic, antipathetic,
hypertensive, hypocholesterolaemia, and hypoglycaemic properties have long been
considered to exist in Moringa species [8].The roots, flowers, gum, and seeds
are commonly used as anti-diabetics and to treat inflammation, heart disease,
liver disease, and haematological, hepatic, and renal function. Moringa leaves,
fruits, and seeds have been shown to be high in protein, important minerals
(calcium, magnesium, potassium, and iron), and vitamins (vitamin A, C and E)
[9]. Plants have recently acquired popularity as a source of new chemicals for
treating microbial diseases. Because of the high cost, low effectiveness, and
developing resistance to conventional treatment, the need for plant-based
antimicrobial is becoming more urgent. Herbal medications are significant in
primary health care in developing countries, particularly where access to
health care is restricted. The moringa plant's natural medicine components have
been shown to decrease tumor incidence in experimental mice [10]. This research
helps to evaluate the nutritional value of Moringa
oleifera during storage periods and microbiological analysis also were
determined. In light of these considerations, the goal of this study was to
evaluate the characteristics of Moringa
oleifera leaf powder in order to expand its use as a functional food
ingredient in food and pharmaceutical products of interest, as well as to
promote this important but underutilized plant that is readily available in
certain parts of the country in a shelf-stable, easily usable form to the
general public.
This research work was
completed at vegetable technology and Food microbiology, Institute of Food
Science and Technology, Bangladesh Council of Scientific and Industrial
Research, Dhaka- 1205.
Sample collection area and processing
Fresh Moringa oleifera (sajna leaves) leaves were collected from the BCSIR campus, Savar, Dhamri and Fulbaria. The sajna leaves were then weighed and properly washed in clean water to remove any clinging soil or dead leaves, as well as to minimize the amount of infected microorganisms. Then, for up to 10 minutes, boil with 800°F water to destroy the enzymes in the food and to prevent unwanted texture changes. Then cooed and 0.1% sodium metabisulphite and were Kept into room temperature for 1 hours. Keep in polybag for further use (Figure 1).
Figure 1: Raw Moringa oleifera leaves.
Methods of nutrient analysis
The nutritional value of
raw and powder product was determined by the following methods.
Determination of moisture
content
Oven drying method
The moisture content of Moringa oleifera samples was measured
using an oven dryer (Wised, Model-msh-30A) that comprises of a chamber in which
trays of Moringa oleifera samples
were put at a controlled temperature (65-75?C) [11]. For three to four days,
drying was started in the oven dryer for 6-7 hours. The amount of Moisture
contents of samples can be calculated by the following procedure.
W1-W2 ×100
% of moisture content = ---------------
W0
W1 =Weight of (sample +
basin) before being dried
W2= Weight of (sample +
basin) after drying
W0 = Weight of sample
Mechanical drying method
The Moringa oleifera sample was dehydrated using a hot air flow
mechanical drier (Wessberg & Tolander Pte. Ltd. Sydney, N.S.W No. 3571).
The dryer is made up of a chamber where product trays are put. A fan blew air
over the trays of items to be dried at 55-600°C, passing through a heater on
the way. An anemometer recorded (600ft/min) air velocity. The amount of drying
was determined by weight loss. Finally make a smooth powder by using a grinder.
The moisture content measured by resulting of weight loss.
Solar drying method
In this research a direct
solar absorption method was applied. Moringa leaves were placed in the chamber.
The heat vaporized the moisture from the leaves. The moisture content measured
by resulting of weight loss.
Determination of Ash (Muffle Furnace)
Weight about 1-2 of
sample and place in a pre-weigh crucible. The sample in a crucible was burned
and transferred to a muffle furnace at 6000C for 4-6 hrs. Ash sample would be
white to slightly grey when ashing was complete. Cool the crucible in desiccators
to room temp and re-weight it. The ash content was calculated as:
W1-W2
×100
% Ash content=
--------------------
Wo
Where,
W1= weight of
the empty crucible
W2= weight of
the crucible + dried sample
Wo= weight of
sample
Protein determination
analysis (Micro-Kjeldahl method)
Digestion
About 0.5 gm of sample
was taken on a ash less filter paper. The sample was transferred to a long neck
kjeldhal flask. Small amount (0.2-0.25 gm) of digestion mixture and 20ml of
conc. H2SO4 was added to the flask and 2 chips were taken in each flask. Heat
the flask over a low flame until the first foaming stops and the liquid boils
vigorously at a moderate rate. Heat for 60 minutes or until the digest is pale
blue in colour. Then cool and transfer the digest to a volumetric flask
(100ml). Then rinse the digestion flask 2 or 3 times with water, cool and make
100 volumes with water.
Distillation and titration
Heated and wash the
kjeldhal apparatus with distilled water. 10ml of 2% boric acid solution was
taken in each 100 ml conical flask and add few drops of mixed indicator which
was placed at narrow tube of kjeldhal apparatus. The tube should be must dipped
into boric acid solution. Introduce 10ml of digestion sample, few drops of
phenolphthalein and 10ml of concentration NaOH through the funnel of kjeldhal
apparatus. Stirs up the digestion mixture and NaOH and librates ammonia which
passes through the condenser and into the boric acid solution as steam. The distilled solution was collected in 100ml
conical flask containing boric acid. After collecting 50ml, the machine was
stopped and the conical flask was removed from the apparatus. The collected
sample from the kjeldhal apparatus was titrated against 0.01N HCL until color
became pink.
Calculation: By using following equation % of nitrogen was
calculated
(S-B)×N×V×100
Nitrogen% = ------------------------
A×W×1000
Where,
S= Titration reading for sample
B= Titration reading for blank
N= Normality of HCL
V= Volume made up the digest
A= Aliquot of the digest taken
W= Weight of the sample
% of protein = % of N2×6.25
Note: 6.25 is the crude
protein factor
Determination of Fat (Soxhlet apparatus
method)
Soxhlet extraction method
was used for the determination of fat. Take 5 gm of sample in a filter paper
bag (Thimble). In the Soxhlet device, place the thimble in the fat extraction
tube. Then connect the extraction tubes bottom to a Soxhlet flask. Load the
correct amount of petroleum ether into the flask through the sample in the
tube. Then connect the condenser to the fat extraction tube and then place in a
water bath. Remove the thimble from the device and distil the majority of the
ether. Pour the ether into a small, dry (already weighed) beaker after it has
achieved a modest volume. Using numerous tiny portions of ether, rinse and filter
the flask completely. On a low heat steam bath, evaporate the ether. Dry at
1000C for 1hr, cool and weigh. The whole process need 8-9 hrs for complete
extraction.
Calculation:
W1-W2×100
% Fat content = -------------------------
Wo
Here
W2=Wt of
beaker+fat
W1=Wt of
beaker
Wo =Wt of
sample.
Determination of
Carbohydrate content (by difference method)
Calculation: Carbohydrate= 100-(moisture+ ash+ protein+ fat+ crude
fiber)
Determination of energy
Calculation: Energy= (protein×4.1) + (fat × 9.3) +
(carbohydrate×4.1)
Methods for microbiological analysis
Three methods such as
pour plate, spread plate and MPN method were used for enumeration of bacteria
and fungus in this research. Serial dilution preparation for pour plate and
spread plate was done followed by proper serial dilution method. 1gm sajna leaf
powder was mixed into 90 ml peptone water then 1 ml sample poured into
petrifies and poured media on it and spread. Then incubate overnight for
bacterial growth. A 0.2ml sample was put onto a solidified agar plate on a
potato dextrose agar (PDA) plate, and the sample was distributed over the agar
plate with the assistance of a sterilized bent glass rod (spreader). There were
5 petriplates for each sample in this study. Yeast and mould counts were
determined by using above two methods [12].
MPN method was used to
enumeration of total coliform bacteria. McCrady (1915), Halvorson, and Zieger
(1933) reported the first precise estimate of the number of live bacteria using
the MPN technique.
Results
Moringa oleifera leaves
were found to be high in proteins and carbohydrates, but low in crude fat,
fiber, and ash, according to these research.
Nutritional composition of raw moringa
olifera leaves
Nutritional composition of raw moringa olifera leaves are presented in the following (Table 1).
Table 1: The chemical compositon of raw Moringa oleifera.
Moisture (%) |
Protein (%) |
Fat (%) |
Fiber (%) |
Ash (%) |
Carbohydrate (%) |
Energy |
81.65 |
8.20 |
2.34 |
1.70 |
2.40 |
3.71 |
70.593 |
Table 2: Effect of storage on solar dried Moringa oleifera leaves powder.
Month |
Moisture |
Protein
|
Fat |
Fiber |
Ash |
Carbohydrate |
Energy |
1st |
3.93 |
15.96 |
9.63 |
8.23 |
2.40 |
59.85 |
400.38 |
2nd |
4.06 |
17.97 |
6.84 |
5.42 |
6.86 |
58.85 |
378.57 |
3rd |
4.11 |
22.90 |
7.23 |
8.16 |
8.69 |
48.91 |
361.66 |
4th |
4.52 |
19.49 |
9.69 |
9.73 |
7.78 |
48.79 |
370.06 |
Table 3: Effect of storage on Mechanical dried Moringa oleifera leaves powder.
Month |
Moisture |
Protein
|
Fat |
Fiber |
Ash |
Carbohydrate |
Energy |
1st
|
7.75 |
34.21 |
3.05 |
6.72 |
6.52 |
58.25 |
407.45 |
2nd |
7.19 |
36.11 |
6.99 |
5.40 |
6.81 |
37.50 |
366.80 |
3rd |
7.86 |
34.31 |
10.70 |
14.19 |
8.32 |
24.62 |
341.12 |
4th |
7.96 |
30.18 |
7.94 |
10.11 |
7.57 |
36.24 |
346.16 |
Table 4: Effect of storage on oven dried Moringa oleifera leaves powder.
Month |
Moisture |
Protein
|
Fat |
Fiber |
Ash |
Carbohydrate |
Energy |
1st |
6.15 |
27.12 |
3.33 |
6.63 |
7.53 |
49.24 |
344.04 |
2nd |
6.28 |
26.14 |
6.40 |
5.48 |
7.45 |
51.75 |
378.86 |
3rd |
6.75 |
24.96 |
9.57 |
11.30 |
7.45 |
39.97 |
355.20 |
4th |
6.82 |
24.17 |
10.35 |
11.89 |
7.45 |
39.32 |
356.56 |
Nutritional composition of powdered product
(Based on Different drying methods)
All data were analyzed
four times for four month to know the time of retention of nutrient of the
powder form of Moringa oleifera
leaves. The data for nutritional composition of solar drying, mechanical drying
and oven drying product were mentioned in table 2, 3 and 4. The observation of
powdered product in different month influenced some of the characteristics of
nutritional composition. It was the effect of dehydration which can vary in
month to month observation (Table 2).
Above table (2) shown
that the gradual increase of moisture ,with the decrease of protein, fat,
fiber, ash, carbohydrate, energy content of leaves powder. When it was freshly
prepared the moisture content was (81.65%). After month to month observation,
the moisture contents were decreased and nutrient contents were increased from
the observation of raw material. For the solar dried Moringa oleifera leaves powder, the moisture contents were
increased and the other nutrient contents were slightly decreased from the
month to month observation (Table 3,4).
From the four month
observation, we can see that some of the nutritional composition were increased
in month to month duration and some of the composition were decreased in month
to month duration and some of them were fluctuated in month to month duration.
This result can variate by the effect of dehydration on the nutritional
composition.
Result of the total bacterial count
In these research plate
count agar (PCA) media were used to calculate the bacterial count from dried
moringa leaves powder. This study represent different bacterial load in
preserved moringa leaves powder. Bacterial count of this study was done by two
categorized like as direct counting from dried sample and count from 10-3
dilution. In the first month, absence of growth in 10-3 dilution of all sample
and presence of growth in others plates where the sample was directly taken
from the dried leaves. In solar dried gradually increased the bacterial counts
and after 4 month TNTC found in 10-3 dilution. In mechanical drier bacterial
count found TNTC after 4th month where oven dried moringa leaves powder found
no growth up to 3rd month of preservation. The total viable bacterial count of
samples are shown in below (Table 5).
Result of the total fungal count
PDA agar media was used for the fungal count of this study by the spread plate methods. In the first and second month absence of growth of fungus in most of the sample and presence of growth of all sample in third and fourth month. Among 3 method oven dried method gives good result.The results are presented in the below Table 6 and Figure 2 (Table 6).
Figure 2: PCA plate with bacterial growth.
Figure 3: PDA plate with fungal growth.
Figure 4: LST media with coliform growth.
Result of the enumeration of total coliform
LST (Laurly Tryptose Sulfate Broth) media by MPN (Most probable Number) three tube method was used for total coliform count and EMB (Eiosin methylene blue) was used to see the fecal contamination. The total coliform count of the samples are shown in Table 7, Figure 3 and 4 .This total coliform counting sample was directly taken from the dried leaves. In the first month observation, absence of coliform and in the second, third and fourth month observation, presence of coliform in all sampling plate (Table 7) (Figure 2-5).
Figure 5: Growth of coliform
bacteria on EMB plate.
Moringa olifera dried powder is very much important now a days in heath sectors. There are so many medicinal value and nutritional value present in dried powder which is helpful for public health. Moringa leaves powder uses as natural products treatment against more than 300 diseases in recent time over the world. In this present study processed powder products were stored in room temperature (25-30?C). The nutritional and microbiological quality of the dried leaves powder were assessed after 6 months of storage. The quality of the processed product was found to be different from that of newly processed products even after one month of storage. The quality of the first month's product was superior to the previous items. It began to degrade after four months of storage and rapidly worsened after five months. The major cause of product degradation was excessive moisture and ascorbic acid levels. As a result, chemical preservatives that are not hazardous to human health should be applied at the required amount allowed by Bangladesh Standard and Testing Institute to extend the shelf-life of processed items. Leaves (84–86%), stems (14–15.5%), and miscellaneous materials were consistently present in Moringa samples (0.25% to 0.40 %).
Table 5: Bacterial count observation of moringa leaves powder on PCA plate.
Month |
Solar Drier |
Mechanical Drier |
Oven Drier | |||