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).