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How to Cite:
Tiwari, S., & Saha, S. N. (2022). Micro filtration of Moringa Oleifera juice using hallow fibre
membrane technology for clarification. International Journal of Health Sciences, 6(S1), 10743–10757.
https://doi.org/10.53730/ijhs.v6nS1.7585
International Journal of Health Sciences ISSN 2550-6978 E-ISSN 2550-696X © 2022.
Manuscript submitted: 27 March 2022, Manuscript revised: 18 April 2022, Accepted for publication: 9 May 2022
10743
Micro filtration of Moringa Oleifera juice using
hallow fibre membrane technology for
clarification
Soumitra Tiwari
Department of Food Processing and Technology, Atal Bihari Vajpayee
Viswavidyalaya, Bilaspur, C.G. PIN 495001
S. N. Saha
Department of Chemical Engineering School of IT, Guru Ghasidas (Central
University) Viswavidyalaya, Bilaspur C.G. PIN 495001
Abstract---Moringa Oleifera plant is growing many countries
including tropical and sub tropical region. It contain numerous
nutrition due to vitamins and proteins. The plant leaf extract having
phenolics and flavanoids such as gallic acids, chlorogenic acid, ferulic
acid, kaempferol, ellagic acid, vanillin and quercetin. The juice is
convenient to herbal health drinks combination of different medicinal
juices, read to use and serve beverages etc. Moringa Oleifera leave
juice has been extracted by grinder then juice were treated with
enzyme and centrifuged. The treated juice were clarified by using
hallow fibre micro membrane for cold pasteurization. The turbidity,
total soluble solid, pH, refractive index, viscosity colour and clarity
were analysed on different treatment. It was concluded that best
results were obtained in terms of Clarity (47.62%T), Turbidity (5.333
NTU), TSS (5.933°B), pH (5.2725%), Viscosity (1.136667 mPa) for
micro filtered juice. The optimum values for the operating condition of
micro filtration were 98.0665 kpa pressures with 0.45 µm pore size.
Keywords---Micro filtration, Moringa Oleifera, hallow fibre.
Introduction
Moringa Oleifera is a native tree plant from Asia and Africa. It is mostly grown in
India, Africa, South America, Mexico and Philippines (Paul & Didia 2012). Now
days the Moringa is used as a rich source of food and food products due to its
considerable inherent nutritional, antioxidant and phytochemical benefits; as well
as its ability to survive in diverse climatic conditions. It has multi-functional
utilities in agriculture, medicine, live stock, human and other biological systems
(Falowo et al 2018, Ndubuaku et al 2015). The trees many parts such as leaf,
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root, bark, seed, flower and pod have been used for boost nutrition and food
securities in developing countries (Popoola et al 2013). The applications of the
leaves are rich sources of vitamins, caroteonoids, polyphenols, phenolic acids,
flavonoids, alkaloids, glucosinolates, isothiocyanates, tannins and saponins
(Leone et al 2015, Sivasankari et al 2014). The leaves have good source vitamin A
(Ferreira et al 2008), Vitamin C (Ramachandran et al 1980), Vitamin E (Borel et al
2013; Efiong et al 2013) and antioxidants (Chambial et al 2013). It is also giving 7
times the Vitamin C of orange, 2 times the protein of yogurt, 4 times the vitamin
A of milk, 3 times the potassium of banana (Gopalakrishnan et al 2016). Moringa
juice is attracting the new food order due to its wide utility in health beverages
and pharmaceutical industry.
Moringa oleifera leaves juice raw juice (R) has a reliving effect in muscular pain
(Bhuvaneshwari et al 2013). The R has many pharmacological functions i.e,
antimicrobial, antifungal, antihypertensive, antihyperlipademic,
antihyperglycemic, antipyretic, wound healing, etc. (Mbikay, 2012; Toma, &
Deyno, 2014). The juice has been persevered mostly through thermal preservation
techniques such as pasteurization and sterilization. During the thermal
preservation the heat sensitive vitamins, polyphenols and others nutrition’s may
lost. So the alternate techniques like membrane processing may require for
preservation of the nutritional rich juices. Membrane processing is a non
thermal green process helpful to retain functional properties present in the juice
(Chemat et al 2017). As per the literature survey the microfiltration has been used
in clarification and purification of the juice for bitter gourd (Jain et al 2018),
banana (Sagu et al 2014), mosambi (Emani et al 2013) etc.
Microfiltration is suitable for separation of micro-organisms, particulates and
there is a free flow of protein, reduction of turbidity (Wagner, 2001), (Al‐Farsi et al
2003) Microfiltration has some benefits on juice clarification without any changes
in the temperature and the pH of the juice and there is no changes in the
chemical constituents of the juice during filtration, improvement in juice quality,
and solves the waste water treatment problem, decreases production and labour
costs (Urošević et al 2017). In this work the fresh Moringa oleifera leaves were
collected from branches, grinded in mixer and convert into viscous paste and then
the viscous paste squeeze in a muslin cloth. The juice obtained is sticky and dark
in colour. The juice was clarified by cellulosic enzyme, centrifuged and the micro
filtrated. The effect of permeate flux and physic-chemical studies such as Total
Soluble Solid (TSS), pH, Viscosity, Turbidity, Colour, Refractive index and Clarity
of the different juice samples observations were recorded.
Materials & Methods:
Preparation of Juice
Fresh and tender leaves are purchased from local market. Then the leaves were
washed with water and then grinded in a mixer grinder (Bajaj Mixer, Bajaj Rex
500-Watt Mixer Grinder, Hyderabad, Telengana, India). The grinded leaves treated
with cellulase enzyme based on the preliminary work with the concentration of
0.1% for 55 min. at 30 C. The obtained juice (R) was used for further treatment.
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Centrifugation
Centrifugation of R juice was performed in a lab scale refrigerated centrifuge (2–
16KL; Sigma-Aldrich). The operation was done on batch mode method with 50 mL
capacity tubes at 25°C. The independent variables for centrifugation operation
were rotational speed (rpm) and time (min). The rotational speed was varied from
6,000 to 8,000 rpm for 20 min and on the basis of preliminary trails. The
supernatant was collected for further physicochemical analysis (total soluble
sugar [TSS], pH, colour value, turbidity, clarity, refractive index, viscosity) after
centrifugation as primary clarified juice (C).
Microfiltration
The R juice performed in a laboratory scale filtration attached with hallow fibre
microfiltration supplied by CleanSep (Mumbai, India). Clarification was carried
out on a tubular microfiltration module using 0.45µm pore size membrane with
0.052m² of permeation area. The membrane was made up of polyether sulfone.
The pressure used was 98.0665 kpa at 30°C. The pressure was kept constant and
the permeate flux was calculated. When the feed passes through the membrane,
filtrate was collected and recirculation of retentate takes place. The filtrate was
measured in measuring cylinder. The permeate flux was calculated by
𝒑𝒆𝒓𝒎𝒆𝒂𝒕𝒆 𝒇𝒍𝒖𝒙 = 𝑽𝒑
𝒕 × 𝑨
Where VP (L) was permeate collected at a certain time period t (h) for the effective
surface area a (m²) of the membrane. The permeate (ECM-M) and retentate (ECM-
R) are evaluated for further physiochemical analysis.
Physiochemical Analysis
Moringa oleifera leaves juice, before and after pre-treatments and microfiltration
processes, were analysed for turbidity, colour, total soluble solids (TSS), viscosity,
pH, clarity, and refractive index.
Viscosity
The viscosity of the juice was dictated by a U-Tub slim viscometer (zenith
glassware, Kolkata, India) at room temperature (28 ± 10°C). The unit of measured
viscosity was represent as mPa.s.
Turbidity
Turbidity was measured with a Digital Turbidity-meter (Model 335; Deluxe
Company, Delhi, India), and the unit of turbidity was nephelometric turbidity
units (NTU).
Colour
Colour was measured as transmittance at 660 nm by a Colorimeter (ColorFlex EZ;
Hunter Lab,eston, Virginia, USA).
∆E= √(∆L ∗)+(∆a ∗)+ (∆b ∗)
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TSS, Refractive index & pH
Abbe-type digital Refractometer was used for TSS and REFERACTIVE INDEX and
expressed in degree Brix (°B).
Clarity
Clarity of the moringaoleifera leaves juice was evaluated by transmittance (%T) at
660 nm utilizing the spectrophotometer (Model: AU 2701; Systronics India, Ltd.,
Ahmedabad, Gujarat, India).
Result &Discussion
Permeate flux:
Clarification was carried out on a tubular microfiltration module using 0.45µm
pore size membrane with 0.052m² of permeation area. The permeate flux was
calculated at constant pressure and temperature were 98.0665 kpa and 300C
respectively. The variation of permeate flux observed with respect time. The initial
filtrate volume was 53 ml in 15 min duration. The filtrate volume was gradually
reduced due to blockage of membrane pore or sedimentation of the solid of high
molecular weight cut off. The permeate flux ranges between 40.76 L/m2.h to
4.616 L/m2.h in 360 minutes. The study state permeates flux achieved after 150
min. The figure 0i; shows the steady state flux increases with respect to time and
after 150 min there are gradual variation on permeate flux. The similar graph
patterns were observed by the Jain et al 2018 in the bitter guard juice during
membrane filtration.
Figure i: Membrane permeate flux variation with respect to time
0
5
10
15
20
25
30
35
40
45
050 100 150 200 250 300 350 400
Permeate Flux
Permeate flux L/m2.h
Time (min)
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TSS (Total Soluble Solids):
The TSS of MLJ ranges from 3.7 to 6.1 ⁰Brix. TSS of enzyme treated (E) and raw
centrifuge juice (C) is higher as compared to raw juice (R). The minimum amount
of soluble solids was present in micro filtrate juice (ECM-F) as compared to raw
juice (R) followed by enzyme treated juice (E) and enzyme treated centrifuge (EC).
TSS is more value due to enzyme concentration was reported by Schobingeret al
1981 in various enzyme treated juices. R has less TSS than enzyme treated juice.
ECM-M has higher TSS as compared to ECM-R. It is observed that juice
undergoing the enzymatic treatment demonstrates higher values of reducing
sugar and TSS during enzymatic hydrolysis. Enzyme treatment is intensified the
degradation of protein and carbohydrates. The same finding is observed by the
Sagu et al 2014 in banana juice. During microfiltration, the turbidity of the
filtrate reduced & reduction of turbidity is also the reason for decrease in TSS.The
reduction of TSS due to microfiltration has been observedby Laorko Aet al 2010 in
pineapple juice and Mirsaeedghazi, H. et al 2010 reported in pomegranate juice.
The TSS of MLJ was estimated after extraction as well as after treatment as
indicated in materials and methods. TSS of MLJ juices after treatment were found
to be significantly lower as compared to raw MLJ. Both the centrifugation and
enzymatic treatment were found to decrease the TSS level in raw MLJ (R).
Centrifugation was found decrease the TSS more as compared to enzymatic
treatment, however, there was no statistical significantdifference in both of them.
Interestingly, their combined treatment (EC) was found to cause a further a highly
significant decrease in TSS level in MLJ. When microfiltration was applied to EC
MLJ, a further significant decrease in TSS level in filtrate was observed. TSS level
of retentate (ECM-R) was estimated and found to be significantly higher as
compared to R.
Figure ii: Total Soluble Solid
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pH :
The MLJ raw juice (R) is acidic in nature. The overall pH value of MLJ different
treated juices ranged between5.25 to 5.57 as described in figure ii. The R has less
pH as compared to E followed by R&C, E&EC. The ECM-R has the lowest pH
content as compared to ECM-F. The pH value of E was higher than R&C&EC. The
growth of microorganisms inhibit due to the low pH. Due to the release of
carboxyl groups and galaturonic acid from pectin, there is decrease of pH values
notify by the Adubofuor, J. et.al 2010. That is desirable changes in juice for long
time preservation. The pH value has a significant role in taste & odour.
Figure iii: pH values of R, E, C, EC, ECM-F & ECM-R
pH level of MLJ before and after vaious treatments was estimated using standard
pH probe method. It was found that enzymatic treatment of MLJ significantly
increases the pH level while centrifugation was found to significantly decrease the
pH level. MLJ after combined EC treatment was found to restores the pH level
deviated by enzymatic or centrifugation treatment and had no significant
difference as compared to R. After using microfiltration of EC treated MLJ, filtrate
and retentate pH level was observed to further decrease in a statistically
significant manner. The pH value has a significant role in taste & odour. Due to
the release of carboxyl groups and galaturonic acid from pectin, there is decrease
of pH values.
Turbidity:
MLJ turbidity was highest of R as compared to E followed by EC, RC. The EC has
less turbidity than R followed by RC&E described in figure 3. The ECM-R has
more turbidity as compared to ECM-F because the juice was clarified by the micro
filtration the same observation was recorded by Mirsaeedghazi, H. et al, 2010 for
10749
pomegranate juice. After micro filtration the clarified juice has low turbidity value.
Low turbidity value gives proper clarified juice. The use of enzyme reduces
turbidity. Less turbidity shows proper clarification of juice. The less turbidity of
juice has more market acceptability.
Figure iv: The MLJ’s R, E, C, EC, ECM-F & ECM-R treated juice turbidity values
in NephelometricTurbidity Units (NTU)
Treatments of MLJ with various methods were found to decrease the turbidity of
MLJ. Turbidity of MLJ after centrifugation or enzymatic treatment was found to
significantly decrease as compared to raw juice. There was a further significant
decrease was observed in the turbidity of MLJ after combined EC treatment. After
microfiltration of EC treated MLJ, turbidity was found to be significantly
decreased in the filtrate. ECM-R retentate was found to have turbidity comparable
with raw MLJ.
Clarity:
Clarity is the one of the prominent characteristics of the juice. The R has the least
clarity than E followed by C, EC, ECM-F & ECM-R. Clarity was highest in micro
filtered juice because after micro filtration the juice becomesclearer. Clarity is the
significant indicator of clarified juice. The clarified juice shows minimum
absorbance as reported by Sin, H. N. et al, 2006 forclarification of sapodilla juice.
10750
Figure v: Clarity in transmittance (%T) of R, E, C, EC, ECM-F & ECM-R
Claritiy of MLJ were estimated and presented in % T. Clarity of raw MLJ was
found to be very low which was found to be significantly augmented after
enzymatic treatment. Centrifugation was found to have a high degree of
clarifyingeffect on MLJ. Combined EC treatmentwas also found to favour the
clarity of MLJ as compared to R or E treatment alone. However, clarity of EC-
treated MLJ was found significantly lower as compared to centrifugation only.
Microfiltration of EC treated MLJ was observed to have the highest clarity among
all the treatment regimens. Interestingly, the retentate after ECM also had a
significantly higher level of clarity as compared to raw MLJ.
Viscosity:
The R juice has higher viscosity as compared to E followed by EC & C. The E&C
have lowest viscosity as compared to R due to high enzyme concentration
decreases the viscosity values are shows in figure V. It is observed that juice
undergoing the enzymatic treatment demonstrates higher value of values of
reducing sugar and TSS during enzymatic hydrolysis. Enzyme treatment is
intensified the degradation of protein and carbohydrates. The findings are similar
to guava juice enzymatic treatment done by Ninga et al 2018.The viscosity value
is highest in ECM-R as compared to ECM-F. Lowest viscosity of juice is more
suitable in enzymatic clarification of juice due to the high viscosity of juice leads
to problem during filtration. It has also observed by the Sin et al 2006 in sapodilla
juice.
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Figure vi: Viscosityin mPaS of R, E, C, EC, ECM-F & ECM-R
Treatments were found to decrease the viscosity of MLJ. Both the enzymatic and
centrifugation treatment was found to significantly decrease the viscosity of MLJ
as compared to R. Among both of these, viscosity level was lower in MLJ after
centrifugation. When combined treatment (EC) was applied, viscosity level of
treated MLJ was found even significantly lower as compared to single enzymatic
or centrifugation treatment. Additional application of microfiltration of EC-treated
MLJ was found to cause statistically significant decrease in viscosity. Viscosity of
ECM-F was found to be least among any of the treatment either alone or in
combination.
Refractive index:
The ECM-R has maximum refractive index 1.3430Brixand the R has minimum
1.330Brix values compared to all types of sample. The sugar content of ECM-R
increases due to microfiltration. The similar finding was observed by Contreras et
al 1992 in the sugar content in various juices. The RI of E,C, EC & ECM-F were
slightly increases in compare to R due to enzymatic treatment on juices. The
figure vi shows the variation of RI on various samples were recorded. There is no
significant variation found on E, C, EC& ECM-F on RI values.
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
R E C EC ECM-F ECM-R
Viscosity (mPAS)
***
***
***
p<0.001
p<0.001
p<0.001
***
**
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Figure vii: Refractive Index in0Brix of R, E, C, EC, ECM-F & ECM-R
RI of raw MLJ and after treatments were estimated and presented in Fig.6. As
indicated, enzymatic and centrifugation treatment were found to augment the RI
of MLJ as compared to raw juice. MLJ obtained after combined EC treatment also
had a significantly higher RI as compared to those of raw juice. Interestingly, MLJ
after combined treatment had a lower RI as compared to single treatment either
enzymatic or centrifugation. However, the difference between RI of MLJ after
combined treatment and singlecentrifugation treatment was found to statistically
significant. The combined EC treatment and enzymatic treatment alone were not
found to significantly differ in their RI level of MLJ. Moreover, additional
microfiltration of EC-treated MLJ was not found to significantly alter the RI level.
Nevertheless, retentate afterECM treatment was found to have significantly higher
level of RI as compared to untreated (raw) or treated MLJ.
1.3300
1.3320
1.3340
1.3360
1.3380
1.3400
1.3420
1.3440
R E C EC ECM-F ECM-R
RI (Brix ˚B)
*** *** **
***
p=0.032
ns
ns
***
10753
Colour
∆E (colour difference) was increased for E as compared to R followed by C, &EC
the observations were shown in figure 7 (a). ∆E (color difference) was increased for
ECM-F as compared to ECM-R. As the observations shown in Fig.7 (b), Chroma
value for E& ECM-F was maximum. As shown in Fig.7(c), Hue value was
maximum negative for R as compared to E followed by EC&C. ECM-R has positive
hue value & negative for ECM-F. ∆E has a qualitative value in a three dimensional
space, Chroma signifies the quantitative attribute characteristics; Hue is the
qualitative representation of the chromatic nature of the colour. The similar
colorvariations were recorded by Ghosh et al 2018 in jamun juice.
Colour differences of MLJ at raw level and after treatment measured through
various method described in materials and methods. As indicated in fig.7(a) and
7(b), colour (L*) and Colour (b) had a similar trend after treatments. Enzymatic
treatment was found to increase both of these colour parameters while
centrifugation had a highly significant but reverse effect. A combined EC
treatment also found to have a decreasing effect on both of these colour
parameters. However, colour (L*) and colour (b) levels of combined treated MLJ
was significantly higher as compared to centrifugation only treatment.
Microfiltration after EC treatment (ECM) was found to further significantly
decrease the colour (L*) and colour (b) level in filtrate. Interestingly, retentate
(ECM-R) had a highercolour (L*) and colour (b) level compared to filtrate (ECM-F),
however, it was found be significantly lower than those of raw MLJ.
Interesting results were obtained for the Hue of the MLJ after treatments (fig.7c).
Raw and enzymatic treated MLJ had negative values. The enzymatic treatment
was found to have significantly less negative value for hue as compared to raw
MLJ. Centrifugation was found to increase the hue of MLJ which was recorded
with positive values. After combined EC treatment of MLJ, hue value was found to
significantly higher as compared to any of the single treatments. However,
additional application of microfiltration was observed to significantly decrease the
hue of MLJ as compared to those of EC treated. Retentate after ECM was found to
have the highest level of hue among any of the treated or raw MLJ.
10754
Figure viii (a): Variations in colour (L*) transmittance in nm of R, E, C, EC, ECM-F
& ECM-R
Figure viii (b): Variations in colour (b) transmittance in nm of R, E, C, EC, ECM-F
& ECM-R
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Figure viii (c): Variations in colour (a) transmittance in nm of R, E, C, EC, ECM-F
& ECM-R
Conclusion
Moringa is thought to have the potential of providing vital nutrition as well as
health and well being to consumers. In this work, fresh Moringa leaves juice
extract was envisaged as a good vehicle of spreadin g its nutraceutical benefits.
Clarification of MLF juice was carried out using both centrifugation and MF.
Comparison of physicochemical properties. It was concluded that best results
were obtained in terms of Clarity (47.62%T), Turbidity (5.333 NTU), Tss(5.933°B),
pH(5.2725%), Viscosity (1.136667mPa)for micro filtered juice. The optimum
values for the operating condition of micro filtration were 98.0665 kpa pressure
with 0.45 µm pore size. Good restoration of the membrane was achieved by
cleaning the membrane with acid and alkali. Fouling was mainly due to the
presence of fibres and macromolecules. Considering quality and quantity, MF is
found to be the most appropriate method for the clarification of MLE juice.
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