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International Journal of Engineering Research and Development
ISSN: 2278-067X, Volume 2, Issue 1 (July 2012), PP. 14-21
www.ijerd.com

          Comparative Study of Different Forms of Moringa Oleifera
                      Extracts for Turbidity Removal
                   Milind R. Gidde1, Anand R Bhalerao2, Chetan N.Malusare3
   1,2
         Professor,Dept. of Civil Engineering, Bharati Vidyapeeth Deemed University College of Engineering, Pune.-411043
             3
               Institute of Environment Education and Research, Bharati Vidyapeeth Deemed University, Pune-411043.



Abstract––Moringa oleifera (M.O) seeds are used as a primary coagulant in drinking water clarification due to presence
of a water–soluble cationic coagulant protein, which is able to reduce the turbidity of treated water. Seeds are powdered
and added to the water directly or after preparing crude extract. The seed kernels contain significant quantities of series
of low molecular weight and water soluble protein, which carries positive charge to the solution. The protein is
considered to act similar to a synthetic and positively charged polymer coagulant. When this protein is added to raw
water, it binds with the predominantly negatively charged particulate making the raw water turbid. Under proper
agitation these bound particulates then grow in size to form the flocks, which may be left to settle by gravity or removed
by sedimentation. The study was carried out to observe the effectiveness of purification M. O. coagulant. The three forms
of coagulants were used viz. Shelled blended, Deoiled powder and Protein powder. At optimum dosage, the percentage
turbidity removal is found to increase with initial turbidity in case of all three coagulants. Among the three forms, protein
powder has more removal efficiency than Deoiled and Shelled blended powder respectively. However, the amount of
protein powder required for coagulation is least, followed by Deoiled and Shelled blended coagulants. Economics of all
these three coagulants is compared with alum (chemical coagulant).

Ke y words––M o ri n g a ol e if e r a, c o ag u l a n t, de o il e d , tu r b i d it y

                                                 I.            INTRODUCTION
          Turbidity removal is one of the important steps in water treatment process and generally is achieved by
coagulation – flocculation – sedimentation process. Common coagulants like alum and iron salts have been widely used in
conventional water treatment processes. Recent studies have pointed out many serious drawbacks of using these coagulants.
Production of large sludge volume, Alzheimer’s disease, reduction in pH and low efficiency of coagulation in low
temperature water are some of the problems faced with these coagulants. Also because of high cost and low availability,
their use is difficult in many developing countries 1. Many researchers have worked on natural coagulants produced and
extracted from plants, animals and microorganisms 2,3. Moringa oleifera is a tropical multipurpose tree that naturally grows
in India, South Africa and South America4. The traditional use of Moringa oleifera seed for domestic household water
treatment is limited to rural areas, where surface water is used for drinking purposes. It has been reported that MO has good
coagulation1,4 and antimicrobial6,8 properties.

           Moringa oleifera seeds are used as a primary coagulant in drinking water clarification and wastewater treatment
due to presence of a water- soluble cationic coagulant protein, which is able to reduce turbidity of treated water. Seeds are
added to the water straight or after preparing crude extract. Seed contains 43% oil, 41% protein and 9% carbohydrates 7.
However, the presence of oil and carbohydrates increases the content of organic matter in the treated water. This fact
represents a disadvantage for its application at full scale water treatment and highly recommends purification of crude
extracts. Extraction of seed oil before crude extract preparation can be a suitable purification option allowing oil recovery for
industrial and food procedures and add value to defatted residues8.

          The seed kernel contains significant quantities of series of low molecular weight and water soluble proteins, which
impart positive charge to solution. The protein is considered to act similar to a synthetic positively charged polymer
coagulant. When this protein is added to raw water, it binds with the predominantly negatively charged particulates which
make raw water turbid. Under proper agitation, the bound particulates then grow in size to form the floc, which may be left
to settle by gravity or may be removed by sedimentation9. MO coagulant protein has been has been identified as low
molecular mass peptide10.

         In the present study shelled blended, deoiled extracts and purified protein powder of Moringa oleifera were used as
coagulant to remove the turbidity. The effects of these different forms of coagulant extracts were studied.

Objective
    1: To study the effect of shelled blended M.O. extract for turbidity removal
    2: To study the effectiveness of deoiled and purified protein powder coagulant for turbidity removal.
    3: To study the cost benefit analysis.



                                                                     14
Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal

                                    II.            MATERIALS AND METHODS
Setting
        This research was carried out in the laboratories of the Department of Civil Engineering at Bharati Vidyapeeth
Deemed University College of Engineering, Pune, India.

Study design
           The Moringa seeds were collected and three forms of these seeds were used as coagulant. We analyzed residual
turbidity after treatment and the optimum dose was determined for varied turbid water samples of 50 NTU, 150 NTU and
450 NTU turbidity.

Preparation of turbid water samples:-
          5 gm of Bentonite clay was mixed to 500 ml distilled water. Mixed clay sample was allowed to soak for 24 hrs.
Suspension was then stirred in the rapid stirrer so as to achieve uniform and homogeneous sample. Resulting suspension was
found to be colloidal and used as stock solution for preparation of turbid water samples. Everyday stock sample of bentonite
clay was diluted by tap water to desired turbidity.

                                           III.           PLANT MATERIAL
Moringa oleifera seeds were collected from surrounding of Pune and Satara districts of Maharashtra. The seeds were
analysed to know the presence of carbohydrates, proteins and oil and other elements (Table No.1).

                                               Table No. 1: Results of Seed analysis
 Sr. No              Sample Name                  Parameters             Results               Units     Test Methods
     1                                               Protein              36.90                 %       AOAC 920.152
     2                                                 Fat                37.25                 %          Ranganna
     3                                           Carbohydrates            16.38                 %        IS: 1656-1997
     4          Moringa oleifera Seeds            Crude Fiber             12.85                 %      SP-18 (P-IX) 1984
     5                                              Moisture              6.41                  %          Ranganna
     6                                                Ash                 3.06                  %        AOAC 940.26

           Pod shells were removed manually and kernel were ground in a domestic blender and sieved through 600µm
stainless steel sieve. The three forms of coagulants were prepared in the following manner.

Coagulant 1: Shelled blended extract
          Tree dried good quality Moringa Oleifera seeds were selected, shelled and the kernel crushed to fine powder in a
blender. Two grams of the powder was added to 100 ml distilled water and blended for 10 minutes at high speed. This
volume was made up to 200 ml resulting in the stock solution with an approximate concentration of 10,000 mg/l (1%).
Fresh stock solution was prepared every day for each day’s experimental run. All experiments were performed with
1% concentration of coagulant.

Coa g u l a n t 2 : De oi l e d p owd e r
           To the crushed shelled blended Moringa oleifera powder, ethanol (95%) was added in 1:10 ratio (1gm of seed
powder and 10ml ethanol) to form a suspension. Then it was mixed with the help of magnetic stirrer for 10 minutes. The
resulting supernatant was separated by centrifugation (300 rpm, 45 min) and the settled material was dried at room
temperature for 24 hours.

Coa g u l a n t 3 : P r ot e i n p owd e r
To ob t a i n t h e pr ot e i n p owd e r fol l owi n g t wo s t e p s wer e p er for me d .

A) Ex t r a ct i o n o f p o lym er
          Dried de-oiled Moringa Oleifera powder was used for extraction of polymer. This powder was then added to 3%
sodium chloride solution. This suspension was continuously agitated for 12 hours in an orbital shaker. The resulting extract
was then filtered through Whatman filter No.44 & brown colored sodium chloride filtrate was collected. Now the filtrate was
further heated in such a way that no white precipitation was formed at the bottom of solution.

B) Pu r if i c at i o n o f Po l ym e r
          Heated crude protein extract solution was further poured into a dialysis tube (Himedia, Mumbai) and kept for 12
hours in a beaker containing cold water which was kept in an ice bath. During dialysis, salts were removed into the
surrounding water solution and white protein remained inside the tube. The protein was removed out from the tube by
rinsing with deionised water. This separated protein was homogenized with cold acetone for delipidization in a homogenizer
to remove lipids. After delipidization this protein was then dried at room temperature.

Coa g u l a n t a c t i vi t y t es t
           Jar tests were performed to determine the effective dosage of coagulant required to reduce the turbidity of the
sample. Jar test was performed using 1 Litre capacity circular baffled jars. The standard procedure consisted 1 minute of
rapid mixing (120 rpm) followed by 15 minutes of slow mixing (30 rpm) for flocculation and 15 minutes of settling.

                                                                  15
Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal

Supernatant was collected from each sample and residual turbidities were used as a base for comparing the efficiency of
coagulation. Jar tests were performed with synthetic turbid water samples of 50 NTU, 150 NTU and 450 NTU turbidity. In
order to ensure the validity of the experiments, all the tests were carefully performed in triplicate and the arithmetic average
values were used for further calculation. The average percentage error of the triplicate samples was 1.2%, with a minimum
of 0.0%, a maximum of 9.5%, and a standard deviation of 2.1% of the goal turbidities of 50 NTU, 150 NTU and 450 NTU.
The actual laboratory initial turbidity values were, on average, within 3.1% of the goal turbidities of 50 NTU, 150 NTU, and
450 NTU. The minimum error from intended initial turbidity was 0.0%, the maximum error was 20.0%, and the standard
deviation was 4.2%. Single factor method of Optimization is used for the study. In this method the experimentation is carried
out by keeping one parameter varying and other parameters are kept constant.The experiments were performed according to
Bureau of Indian Standards IS 3025(PART 50): 2001

                                                               IV.      COST ECONOMICS
          Each tree can produce approximately 400-1000 pods per year i.e. 15000 to 25000 seeds per year11. The average
weight of non shelled seed is 300 mg. The kernel to hull ratio is 75.25%.

         To compare the cost of Moringa seeds with that of alum, a case study of one village was studied. For one village
with a population of 5000, water requirement is 200,000 lit/day.For daily water treatment, required quantity of Alum and
Moringa coagulants and correspondingly plantation to produce required Moringa seeds for three coagulants are given in
Table No. 2. The area required for plantation of one M.O. tree is considered to be 9 sq. m.

                                                          V.         RESULTS AND DISCUSSION
         Jar test experiments with different Moringa oleifera coagulant forms were performed by using three forms of
coagulant extracts. These coagulants were prepared by using a standard preparation method as discussed earlier. Initially the
optimum dosage was determined. This was the dosage of coagulant corresponding to the lowest residual turbidity.At
optimum dosage, the percent turbidity removal rate was found to increase with initial turbidity in case of all three coagulants.

1) Shelled blended Moringa oleifera coagulant was able to achieve 76.45% turbidity removal at an optimum dosage of 70
mg/L for 50 NTU whilst for 150 and 450 NTU turbidity the dosage were 120 mg/L and 240 mg/L respectively and
percentage turbidity removal was 92.33% and 97.7% respectively.(Graph-1,2,3)


                                                          30
                                                                                       70 mg/l Optimum…
                                                          25
                               Residual terbidity (NTU)




                                                                         24       23 21
                                                          20
                                                                                         20
                                                          15
                                                                                                           12.7
                                                                                              11.8
                                                          10
                                                           5
                                                           0
                                                                0                50              100
                                                                M O Shelled blended Dose (mg/L)
                                                               Graph No.1 - Optimum dose for 50…
                                                          35
                                                                           30
                                                          30
                               Residual terbidity (NTU)




                                                                                  27
                                                          25                            22.1
                                                          20
                                                                                               19          16
                                                          15
                                                          10
                                                                      120 mg/l                      11.2
                                                           5          Optimum …
                                                           0
                                                               50    70      90    110     130    150
                                                                 M O Shelled blended Dose (mg/L)
                                                                Graph No.2- Optimum dose for 150…


                                                                                16
Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal


                                               35
                                                                                                          33          30
                                               30


                    Residual terbidity (NTU)
                                                                                                                           27
                                               25
                                               20                                                                                21
                                                                                                                                             17.1
                                               15
                                                                                                                                      12.7
                                               10
                                                                                          240 mg/l
                                               5                                          Optimum
                                                                                          dose
                                               0
                                                    150    170     190     210     230     250     270
                                                           M O Shelled blended Dose (mg/L)
                                                    Graph No.3- Optimum dose for 450 NTU turbidity


2) Deoiled powder of Moringa oleifera was able to achieve 77.8% turbidity removal at an optimum dosage of 50 mg/L for
50 NTU whilst for 150 and 450 NTU turbidity the dosage were 90 mg/L and 210 mg/L respectively and percentage turbidity
removal was 95.86% and 98.11% respectively (Graph-4,5,6).
                                                                           30
                                                                                                                            50 mg/l
                                                                           25                                               Optimum dose
                                                                                             24
                                                      Residual terbidity (NTU)




                                                                                                     20
                                                                           20
                                                                                                            17
                                                                           15
                                                                                                                13.1              14.5
                                                                           10
                                                                                                                           11
                                                                                 5

                                                                                 0
                                                                                      0           20        40      60        80
                                                                                              M O Deoiled powder Dose (mg/L)
                                                                                           Graph No.4 - Optimum dose for 50 NTU
                                                                                                         turbidity
                                                                                 12
                                                                                                     11.4
                                                                                 10
                                                                                                               8.2
                                                    Residual terbidity (NTU)




                                                                                                                                 9.4 9.8
                                                                                 8
                                                                                                                           7.4
                                                                                 6
                                                                                                                     6.2
                                                                                 4

                                                                                 2                                               90 mg/l
                                                                                                                                 Optimum …
                                                                                 0
                                                                                      50         70         90     110       130
                                                                                             M O Deoiled powder Dose (mg/L)
                                                                                          Graph No.5 - Optimum dose for 150 NTU
                                                                                                         turbidity




                                                                                                           17
Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal


                                                 12
                                                                                                    11.4
                                                 10
                                                                                                           8.2


                      Residual terbidity (NTU)
                                                                                                                                       9.4     9.8
                                                 8
                                                                                                                               7.4
                                                 6
                                                                                                                   6.2
                                                 4
                                                                                                                                    90 mg/l
                                                 2                                                                                  Optimum dose
                                                 0
                                                       50                                      70                 90                110              130
                                                                                     M O Deoiled powder Dose (mg/L)
                                                                                  Graph No.5 - Optimum dose for 150 NTU
                                                                                                 turbidity

3) Protein powder extract of Moringa oleifera was able to achieve 84.4% turbidity removal at an optimum dosage of 20
mg/L for 50 NTU whilst for 150 and 450 NTU turbidity the dosage were 40 mg/L and 100 mg/L respectively and percentage
turbidity removal was 95.86% and 98.11% respectively.(Graph-7,8,9)
                                                                    14
                                                                                                                                12.7
                                                                    12
                                                                                                                 10.7
                                                      Residual terbidity (NTU)




                                                                    10
                                                                                             9.1
                                                                                  8
                                                                                                           7.8
                                                                                  6
                                                                                  4
                                                                                                                                     20 mg/l
                                                                                  2                                                  Optimum
                                                                                                                                     dose
                                                                                  0
                                                                                      0            20             40         60
                                                                                            M O Protein powder Dose (mg/L)
                                                                                          Graph No.7- Optimum dose for 50 NTU
                                                                                                        turbidity
                                                                     14
                                                                     12                                      12.5
                                                                                                                                     10.2
                                                       Residual terbidity (NTU)




                                                                     10
                                                                                                                              9.7
                                                                                  8
                                                                                  6
                                                                                                                        5.5
                                                                                  4
                                                                                                                                     40 mg/l
                                                                                  2                                                  Optimum
                                                                                                                                     dose
                                                                                  0
                                                                                      0         20         40      60       80
                                                                                           M O Protein powder Dose (mg/L)
                                                                                          Graph No.8 - Optimum dose for 150
                                                                                                    NTU turbidity



                                                                                                            18
Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal

                                                      16
                                                                                                            14.1
                                                      14
                                                      12




                           Residual terbidity (NTU)
                                                      10
                                                                                                                                              9.2
                                                           8
                                                                                                                     6.7                7.7
                                                           6
                                                           4
                                                                                                                                          100 mg/l
                                                           2                                                                              Optimum
                                                           0                                                                              dose

                                                                             50                70       90     110     130                           150
                                                                                            M O Protein powder Dose (mg/L)
                                                                                          Graph No. 9- Optimum dose for 450…

          The oil content in the seed forms an emulsion or film coating, which may inhibit the contact with surface of
reaction and thus reduce flocculation. This can be a possible explanation for maximum percentage of turbidity reduction
when coagulant has no oil content.

Percentage reduction in dose
          At 50 NTU turbidity, 28.57% dose reduction in de-oiled powder and 71.42% reduction in protein powder were
observed whereas for 150 NTU turbidity, the percentage reduction in dose for de-oiled powder was observed to be 25% and
for protein powder was 66.66%. Whereas for 450 NTU turbidity, the percentage reduction in dose for de-oiled powder was
observed to be 12.5% and for protein powder was 58.38 % (Graph-10, 11, 12).

                                                                                                Initial turbidity 50 NTU percentage
                                                                   percentage reduction




                                                                                                          reduction in dose
                                                                                          70                                58.38
                                                                                          60
                                                                                          50
                                                                                          40
                                                                                          30            12.5
                                                                                          20
                                                                                          10
                                                                                           0
                                                                                                  Shelled blended-     Shelled blended-
                                                                                                  Deoiled powder        protein powder
                                                                                                Graph No.10-M.O.powder


                                                                                               Initial turbidity 150 NTU percentage
                                                      percentage reduction




                                                                                                          reduction in dose
                                                                             70                                       66.66
                                                                             60
                                                                             50
                                                                             40                      25
                                                                             30
                                                                             20
                                                                             10
                                                                              0
                                                                                               Shelled blended-      Shelled blended-
                                                                                               Deoiled powder         protein powder
                                                                                                 Graph No.11- M.O.powder




                                                                                                                  19
Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal


                                                                  Initial turbidity 450 NTU percentage




                                      percentage reduction
                                                                             reduction in dose
                                                             80                           71
                                                             70
                                                             60
                                                             50
                                                             40         28.57
                                                             30
                                                             20
                                                             10
                                                              0
                                                                  Shelled blended-    Shelled blended-
                                                                  Deoiled powder       protein powder

                                                                   Graph No.12- M.O.powder


          Shelled blended, de-oiled powder & protein powder, all three are being used as coagulants. Among them, protein
powder has more removal efficiency than de-oiled & shelled blended powder respectively, but the amount of protein powder
as coagulant requirement is less followed by de-oiled & Shelled blended.

Cost economics
          For cost economics studies one village having population 5000 was considered. Water requirement for this village
@ 40 lpcd is 2 lakh litres per day. For daily water treatment the amount of coagulant required depends upon the turbidity of
water in that area. If a village receives turbid water of 50 NTU, then optimum dose of alum is 30 mg/lit. So the quantity of
alum required will be 6 kg per day, which will cost nearly Rs.96 per day at the rate of Rs.16/kg. Whereas in case of different
forms of Moringa coagulant - shelled blended, deoiled powder, protein powder - the optimum dose is 70 mg/lit, 50 mg/lit &
30 mg/lit respectively. The total quantity of these coagulants required per day will be 14 kg of shelled blended, 10 kg of
deoiled powder & 2.7 kg of protein powder. It will cost around Rs.2800 for shelled blended & Rs.1200 for deoiled powder
per day. Then the area required for cultivation of Moringa plantation was determined. Then to obtain required quantity of
coagulant (shelled blended, deoiled powder, protein powder), the number of trees is 637, 775 and 342 respectively and the
land requirement for cultivation is 1.42 acre, 1.72 acre & 0.76 acre respectively (for 150 NTU and 450 NTU turbidity water,
the required coagulant cost and number of trees are given in the Table No.2.

                                                                  Ta b l e No .2 : Cos t Ec on omi c s
 Form of Coagulant              Initial                           Optimum             Seed       Rate        Cost         No of         Area
                               Turbidity                          dose (mg/l)      Quantity      (Rs)      (Rs/day)       trees        (acre)
                                                                                                                          (182. day)
                                (NTU)                                               (kg/day)
 Shelled blended                  50                                  70               14         Rs.       2800            637        1.42
                                                                                                200/kg
                                   150                               120               24                   4800           1092        2.43
                                   450                                  240               48                9600           2184        4.86
 Deoiled powder                    50                                   50                10       Rs.      1200            775        1.72
                                                                                                 120/kg.
                                   150                                  90                18                2160           1368        3.04
                                   450                                  210               42                5040           3193         7.1
 Protein powder                    50                                   20                2.7              Data not         342        0.76
                                                                                                           available
                                   150                                  40                5.4                               664         1.4
                                   450                                  100           13.5                                 1642        3.65
 Alum                              50                                   30                6        Rs.                 Not required
                                                                                                  16/kg.
                                   150                                  50                10
                                   450                                  90                18
( S ou r ce s - * Ri mp l e a l oe ve r a p r od u ct s , Gu j a ra t * Rahe j a ch e mi ca l s , P u n e , * Nat i on a l r u r al dr i n ki n g
wa t e r pr ogr a m, * J a h n , 1 9 8 8, * P a r a g S a d d gi r, 2 0 0 7 )

                                                                  VI.           CONCLUSION
          Three coagulants shelled blended powder; deoiled powder and protein powder of the Moringa oleifera coagulant
were used in the present study. Among them, protein powder has maximum removal efficiency compared to deoiled and
shelled blended powder. The Shelled blended powder contains carbohydrates, oil and protein. The protein acts as a
coagulant. The oil and carbohydrates present increase the organic matter contained in the treated water. This makes water
unsuitable for drinking after storing it for some time. The deoiled powder contains protein and carbohydrates. The removal

                                                                                     20
Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal

of oil reduces the organic matter content of the water thus making water more suitable for drinking. The purified proteins
reduce the organic matter content to the least. The reduction in the optimum dose required for shelled blended powder to
deoiled powder to protein powder is because of the removal of oil and the carbohydrates, as only protein acts as a coagulant.

          In many villages of India, hundreds of acres of barren land are available. These lands can be utilized for cultivation
of Moringa plants. As this plant does not require any specific soil type to grow, it can grow even in semi-arid regions as
well. This plantation ultimately reduces the cost of chemical coagulants and also different byproducts of Moringa like leaf
powder, Moringa oil and flowers are available. It can be of better economic benefits to locals.

                                     VII.             ACKNOWLEDGEMENT
         The authors are grateful to Bharati Vidyapeeth Institute of Environment Education & Research Pune, and
B.V.D.U. College of Engineering, Pune and Rajiv Gandhi Institute of I.T. & B.T. Pune.

                                                        REFERENCES
  [1].      Ndabigengesere Anselme, Narasiah K.S Quality of water treated by coagulation using Moringa Oleifera seeds. Water Res. 32(3),
            781-791(1998)
  [2].      Kawamura S. Effectiveness of natural polyelectrolytes in water treatment. JAWWA, 83(10), 88-91 (1991)
  [3].      Lee S.H., Lee S. O., Jang K.L., Lee T. H.,Microbial flocculant from arcuadendron SP – 49. Biotechnol. Lett. 17(1), 95 – 100
            (2001)
  [4].      Jahn S.A.A. Using Moringa seeds as coagulants in developing countries. Journal of American Water Works Association, 80:43–
            50 (1988).
  [5].      Muyibi S.A. and Evison L.M Optimising Physical Parameters affecting Coagulation of Turbid water with Moringa Oleifera
            seeds. Water Res. 29 (12), 2689-2695 (1995)
  [6].      Olsen Annette Low technology water purification by Bentonite clay and Moringa oleifera seed flocculation as performed in
            Sudanese villages: Effects on Schistosoma mansoni cercariae. Water Res. 21 (5) 517-522(1987)
  [7].      Anhwange B.A., Ajibola V.O., Oniye S.J. Chemical studies of the seeds of Moringa oleifera (Lam) and Detarium microcarpum
            (Guill and Spurr) J. Of Biological Sciences 4(6) 711-715 (2004)
  [8].      Ghebremichael K.A, Gunaratna K.R, Henriksson H,Brumer H, Dalhammar G. A. Simple purification and activity assay of the
            coagulant protein from Moringa oleifera Seed. Water Research 39: 2338-2344 (2005).
  [9].      Geoff Folkard, John Sutherland, Rod Shaw .water clarification using Moringa oleifera seed as coagulant. Val-3, (2007)
  [10]. Gassenschmidt U, Jany K K, et al. Isolation and characterization of a flocculation protein from MO lam. BBA Biochem Biophys
            Acta 1243: 477-481 (1995)
  [ 1 1 ] . Amelia Pilar Rauter .Natural products in new millennium prospects and Indusry, page no-405-416, (2002).




                                                                 21

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Moringa extracts study turbidity removal

  • 1. International Journal of Engineering Research and Development ISSN: 2278-067X, Volume 2, Issue 1 (July 2012), PP. 14-21 www.ijerd.com Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal Milind R. Gidde1, Anand R Bhalerao2, Chetan N.Malusare3 1,2 Professor,Dept. of Civil Engineering, Bharati Vidyapeeth Deemed University College of Engineering, Pune.-411043 3 Institute of Environment Education and Research, Bharati Vidyapeeth Deemed University, Pune-411043. Abstract––Moringa oleifera (M.O) seeds are used as a primary coagulant in drinking water clarification due to presence of a water–soluble cationic coagulant protein, which is able to reduce the turbidity of treated water. Seeds are powdered and added to the water directly or after preparing crude extract. The seed kernels contain significant quantities of series of low molecular weight and water soluble protein, which carries positive charge to the solution. The protein is considered to act similar to a synthetic and positively charged polymer coagulant. When this protein is added to raw water, it binds with the predominantly negatively charged particulate making the raw water turbid. Under proper agitation these bound particulates then grow in size to form the flocks, which may be left to settle by gravity or removed by sedimentation. The study was carried out to observe the effectiveness of purification M. O. coagulant. The three forms of coagulants were used viz. Shelled blended, Deoiled powder and Protein powder. At optimum dosage, the percentage turbidity removal is found to increase with initial turbidity in case of all three coagulants. Among the three forms, protein powder has more removal efficiency than Deoiled and Shelled blended powder respectively. However, the amount of protein powder required for coagulation is least, followed by Deoiled and Shelled blended coagulants. Economics of all these three coagulants is compared with alum (chemical coagulant). Ke y words––M o ri n g a ol e if e r a, c o ag u l a n t, de o il e d , tu r b i d it y I. INTRODUCTION Turbidity removal is one of the important steps in water treatment process and generally is achieved by coagulation – flocculation – sedimentation process. Common coagulants like alum and iron salts have been widely used in conventional water treatment processes. Recent studies have pointed out many serious drawbacks of using these coagulants. Production of large sludge volume, Alzheimer’s disease, reduction in pH and low efficiency of coagulation in low temperature water are some of the problems faced with these coagulants. Also because of high cost and low availability, their use is difficult in many developing countries 1. Many researchers have worked on natural coagulants produced and extracted from plants, animals and microorganisms 2,3. Moringa oleifera is a tropical multipurpose tree that naturally grows in India, South Africa and South America4. The traditional use of Moringa oleifera seed for domestic household water treatment is limited to rural areas, where surface water is used for drinking purposes. It has been reported that MO has good coagulation1,4 and antimicrobial6,8 properties. Moringa oleifera seeds are used as a primary coagulant in drinking water clarification and wastewater treatment due to presence of a water- soluble cationic coagulant protein, which is able to reduce turbidity of treated water. Seeds are added to the water straight or after preparing crude extract. Seed contains 43% oil, 41% protein and 9% carbohydrates 7. However, the presence of oil and carbohydrates increases the content of organic matter in the treated water. This fact represents a disadvantage for its application at full scale water treatment and highly recommends purification of crude extracts. Extraction of seed oil before crude extract preparation can be a suitable purification option allowing oil recovery for industrial and food procedures and add value to defatted residues8. The seed kernel contains significant quantities of series of low molecular weight and water soluble proteins, which impart positive charge to solution. The protein is considered to act similar to a synthetic positively charged polymer coagulant. When this protein is added to raw water, it binds with the predominantly negatively charged particulates which make raw water turbid. Under proper agitation, the bound particulates then grow in size to form the floc, which may be left to settle by gravity or may be removed by sedimentation9. MO coagulant protein has been has been identified as low molecular mass peptide10. In the present study shelled blended, deoiled extracts and purified protein powder of Moringa oleifera were used as coagulant to remove the turbidity. The effects of these different forms of coagulant extracts were studied. Objective 1: To study the effect of shelled blended M.O. extract for turbidity removal 2: To study the effectiveness of deoiled and purified protein powder coagulant for turbidity removal. 3: To study the cost benefit analysis. 14
  • 2. Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal II. MATERIALS AND METHODS Setting This research was carried out in the laboratories of the Department of Civil Engineering at Bharati Vidyapeeth Deemed University College of Engineering, Pune, India. Study design The Moringa seeds were collected and three forms of these seeds were used as coagulant. We analyzed residual turbidity after treatment and the optimum dose was determined for varied turbid water samples of 50 NTU, 150 NTU and 450 NTU turbidity. Preparation of turbid water samples:- 5 gm of Bentonite clay was mixed to 500 ml distilled water. Mixed clay sample was allowed to soak for 24 hrs. Suspension was then stirred in the rapid stirrer so as to achieve uniform and homogeneous sample. Resulting suspension was found to be colloidal and used as stock solution for preparation of turbid water samples. Everyday stock sample of bentonite clay was diluted by tap water to desired turbidity. III. PLANT MATERIAL Moringa oleifera seeds were collected from surrounding of Pune and Satara districts of Maharashtra. The seeds were analysed to know the presence of carbohydrates, proteins and oil and other elements (Table No.1). Table No. 1: Results of Seed analysis Sr. No Sample Name Parameters Results Units Test Methods 1 Protein 36.90 % AOAC 920.152 2 Fat 37.25 % Ranganna 3 Carbohydrates 16.38 % IS: 1656-1997 4 Moringa oleifera Seeds Crude Fiber 12.85 % SP-18 (P-IX) 1984 5 Moisture 6.41 % Ranganna 6 Ash 3.06 % AOAC 940.26 Pod shells were removed manually and kernel were ground in a domestic blender and sieved through 600µm stainless steel sieve. The three forms of coagulants were prepared in the following manner. Coagulant 1: Shelled blended extract Tree dried good quality Moringa Oleifera seeds were selected, shelled and the kernel crushed to fine powder in a blender. Two grams of the powder was added to 100 ml distilled water and blended for 10 minutes at high speed. This volume was made up to 200 ml resulting in the stock solution with an approximate concentration of 10,000 mg/l (1%). Fresh stock solution was prepared every day for each day’s experimental run. All experiments were performed with 1% concentration of coagulant. Coa g u l a n t 2 : De oi l e d p owd e r To the crushed shelled blended Moringa oleifera powder, ethanol (95%) was added in 1:10 ratio (1gm of seed powder and 10ml ethanol) to form a suspension. Then it was mixed with the help of magnetic stirrer for 10 minutes. The resulting supernatant was separated by centrifugation (300 rpm, 45 min) and the settled material was dried at room temperature for 24 hours. Coa g u l a n t 3 : P r ot e i n p owd e r To ob t a i n t h e pr ot e i n p owd e r fol l owi n g t wo s t e p s wer e p er for me d . A) Ex t r a ct i o n o f p o lym er Dried de-oiled Moringa Oleifera powder was used for extraction of polymer. This powder was then added to 3% sodium chloride solution. This suspension was continuously agitated for 12 hours in an orbital shaker. The resulting extract was then filtered through Whatman filter No.44 & brown colored sodium chloride filtrate was collected. Now the filtrate was further heated in such a way that no white precipitation was formed at the bottom of solution. B) Pu r if i c at i o n o f Po l ym e r Heated crude protein extract solution was further poured into a dialysis tube (Himedia, Mumbai) and kept for 12 hours in a beaker containing cold water which was kept in an ice bath. During dialysis, salts were removed into the surrounding water solution and white protein remained inside the tube. The protein was removed out from the tube by rinsing with deionised water. This separated protein was homogenized with cold acetone for delipidization in a homogenizer to remove lipids. After delipidization this protein was then dried at room temperature. Coa g u l a n t a c t i vi t y t es t Jar tests were performed to determine the effective dosage of coagulant required to reduce the turbidity of the sample. Jar test was performed using 1 Litre capacity circular baffled jars. The standard procedure consisted 1 minute of rapid mixing (120 rpm) followed by 15 minutes of slow mixing (30 rpm) for flocculation and 15 minutes of settling. 15
  • 3. Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal Supernatant was collected from each sample and residual turbidities were used as a base for comparing the efficiency of coagulation. Jar tests were performed with synthetic turbid water samples of 50 NTU, 150 NTU and 450 NTU turbidity. In order to ensure the validity of the experiments, all the tests were carefully performed in triplicate and the arithmetic average values were used for further calculation. The average percentage error of the triplicate samples was 1.2%, with a minimum of 0.0%, a maximum of 9.5%, and a standard deviation of 2.1% of the goal turbidities of 50 NTU, 150 NTU and 450 NTU. The actual laboratory initial turbidity values were, on average, within 3.1% of the goal turbidities of 50 NTU, 150 NTU, and 450 NTU. The minimum error from intended initial turbidity was 0.0%, the maximum error was 20.0%, and the standard deviation was 4.2%. Single factor method of Optimization is used for the study. In this method the experimentation is carried out by keeping one parameter varying and other parameters are kept constant.The experiments were performed according to Bureau of Indian Standards IS 3025(PART 50): 2001 IV. COST ECONOMICS Each tree can produce approximately 400-1000 pods per year i.e. 15000 to 25000 seeds per year11. The average weight of non shelled seed is 300 mg. The kernel to hull ratio is 75.25%. To compare the cost of Moringa seeds with that of alum, a case study of one village was studied. For one village with a population of 5000, water requirement is 200,000 lit/day.For daily water treatment, required quantity of Alum and Moringa coagulants and correspondingly plantation to produce required Moringa seeds for three coagulants are given in Table No. 2. The area required for plantation of one M.O. tree is considered to be 9 sq. m. V. RESULTS AND DISCUSSION Jar test experiments with different Moringa oleifera coagulant forms were performed by using three forms of coagulant extracts. These coagulants were prepared by using a standard preparation method as discussed earlier. Initially the optimum dosage was determined. This was the dosage of coagulant corresponding to the lowest residual turbidity.At optimum dosage, the percent turbidity removal rate was found to increase with initial turbidity in case of all three coagulants. 1) Shelled blended Moringa oleifera coagulant was able to achieve 76.45% turbidity removal at an optimum dosage of 70 mg/L for 50 NTU whilst for 150 and 450 NTU turbidity the dosage were 120 mg/L and 240 mg/L respectively and percentage turbidity removal was 92.33% and 97.7% respectively.(Graph-1,2,3) 30 70 mg/l Optimum… 25 Residual terbidity (NTU) 24 23 21 20 20 15 12.7 11.8 10 5 0 0 50 100 M O Shelled blended Dose (mg/L) Graph No.1 - Optimum dose for 50… 35 30 30 Residual terbidity (NTU) 27 25 22.1 20 19 16 15 10 120 mg/l 11.2 5 Optimum … 0 50 70 90 110 130 150 M O Shelled blended Dose (mg/L) Graph No.2- Optimum dose for 150… 16
  • 4. Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal 35 33 30 30 Residual terbidity (NTU) 27 25 20 21 17.1 15 12.7 10 240 mg/l 5 Optimum dose 0 150 170 190 210 230 250 270 M O Shelled blended Dose (mg/L) Graph No.3- Optimum dose for 450 NTU turbidity 2) Deoiled powder of Moringa oleifera was able to achieve 77.8% turbidity removal at an optimum dosage of 50 mg/L for 50 NTU whilst for 150 and 450 NTU turbidity the dosage were 90 mg/L and 210 mg/L respectively and percentage turbidity removal was 95.86% and 98.11% respectively (Graph-4,5,6). 30 50 mg/l 25 Optimum dose 24 Residual terbidity (NTU) 20 20 17 15 13.1 14.5 10 11 5 0 0 20 40 60 80 M O Deoiled powder Dose (mg/L) Graph No.4 - Optimum dose for 50 NTU turbidity 12 11.4 10 8.2 Residual terbidity (NTU) 9.4 9.8 8 7.4 6 6.2 4 2 90 mg/l Optimum … 0 50 70 90 110 130 M O Deoiled powder Dose (mg/L) Graph No.5 - Optimum dose for 150 NTU turbidity 17
  • 5. Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal 12 11.4 10 8.2 Residual terbidity (NTU) 9.4 9.8 8 7.4 6 6.2 4 90 mg/l 2 Optimum dose 0 50 70 90 110 130 M O Deoiled powder Dose (mg/L) Graph No.5 - Optimum dose for 150 NTU turbidity 3) Protein powder extract of Moringa oleifera was able to achieve 84.4% turbidity removal at an optimum dosage of 20 mg/L for 50 NTU whilst for 150 and 450 NTU turbidity the dosage were 40 mg/L and 100 mg/L respectively and percentage turbidity removal was 95.86% and 98.11% respectively.(Graph-7,8,9) 14 12.7 12 10.7 Residual terbidity (NTU) 10 9.1 8 7.8 6 4 20 mg/l 2 Optimum dose 0 0 20 40 60 M O Protein powder Dose (mg/L) Graph No.7- Optimum dose for 50 NTU turbidity 14 12 12.5 10.2 Residual terbidity (NTU) 10 9.7 8 6 5.5 4 40 mg/l 2 Optimum dose 0 0 20 40 60 80 M O Protein powder Dose (mg/L) Graph No.8 - Optimum dose for 150 NTU turbidity 18
  • 6. Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal 16 14.1 14 12 Residual terbidity (NTU) 10 9.2 8 6.7 7.7 6 4 100 mg/l 2 Optimum 0 dose 50 70 90 110 130 150 M O Protein powder Dose (mg/L) Graph No. 9- Optimum dose for 450… The oil content in the seed forms an emulsion or film coating, which may inhibit the contact with surface of reaction and thus reduce flocculation. This can be a possible explanation for maximum percentage of turbidity reduction when coagulant has no oil content. Percentage reduction in dose At 50 NTU turbidity, 28.57% dose reduction in de-oiled powder and 71.42% reduction in protein powder were observed whereas for 150 NTU turbidity, the percentage reduction in dose for de-oiled powder was observed to be 25% and for protein powder was 66.66%. Whereas for 450 NTU turbidity, the percentage reduction in dose for de-oiled powder was observed to be 12.5% and for protein powder was 58.38 % (Graph-10, 11, 12). Initial turbidity 50 NTU percentage percentage reduction reduction in dose 70 58.38 60 50 40 30 12.5 20 10 0 Shelled blended- Shelled blended- Deoiled powder protein powder Graph No.10-M.O.powder Initial turbidity 150 NTU percentage percentage reduction reduction in dose 70 66.66 60 50 40 25 30 20 10 0 Shelled blended- Shelled blended- Deoiled powder protein powder Graph No.11- M.O.powder 19
  • 7. Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal Initial turbidity 450 NTU percentage percentage reduction reduction in dose 80 71 70 60 50 40 28.57 30 20 10 0 Shelled blended- Shelled blended- Deoiled powder protein powder Graph No.12- M.O.powder Shelled blended, de-oiled powder & protein powder, all three are being used as coagulants. Among them, protein powder has more removal efficiency than de-oiled & shelled blended powder respectively, but the amount of protein powder as coagulant requirement is less followed by de-oiled & Shelled blended. Cost economics For cost economics studies one village having population 5000 was considered. Water requirement for this village @ 40 lpcd is 2 lakh litres per day. For daily water treatment the amount of coagulant required depends upon the turbidity of water in that area. If a village receives turbid water of 50 NTU, then optimum dose of alum is 30 mg/lit. So the quantity of alum required will be 6 kg per day, which will cost nearly Rs.96 per day at the rate of Rs.16/kg. Whereas in case of different forms of Moringa coagulant - shelled blended, deoiled powder, protein powder - the optimum dose is 70 mg/lit, 50 mg/lit & 30 mg/lit respectively. The total quantity of these coagulants required per day will be 14 kg of shelled blended, 10 kg of deoiled powder & 2.7 kg of protein powder. It will cost around Rs.2800 for shelled blended & Rs.1200 for deoiled powder per day. Then the area required for cultivation of Moringa plantation was determined. Then to obtain required quantity of coagulant (shelled blended, deoiled powder, protein powder), the number of trees is 637, 775 and 342 respectively and the land requirement for cultivation is 1.42 acre, 1.72 acre & 0.76 acre respectively (for 150 NTU and 450 NTU turbidity water, the required coagulant cost and number of trees are given in the Table No.2. Ta b l e No .2 : Cos t Ec on omi c s Form of Coagulant Initial Optimum Seed Rate Cost No of Area Turbidity dose (mg/l) Quantity (Rs) (Rs/day) trees (acre) (182. day) (NTU) (kg/day) Shelled blended 50 70 14 Rs. 2800 637 1.42 200/kg 150 120 24 4800 1092 2.43 450 240 48 9600 2184 4.86 Deoiled powder 50 50 10 Rs. 1200 775 1.72 120/kg. 150 90 18 2160 1368 3.04 450 210 42 5040 3193 7.1 Protein powder 50 20 2.7 Data not 342 0.76 available 150 40 5.4 664 1.4 450 100 13.5 1642 3.65 Alum 50 30 6 Rs. Not required 16/kg. 150 50 10 450 90 18 ( S ou r ce s - * Ri mp l e a l oe ve r a p r od u ct s , Gu j a ra t * Rahe j a ch e mi ca l s , P u n e , * Nat i on a l r u r al dr i n ki n g wa t e r pr ogr a m, * J a h n , 1 9 8 8, * P a r a g S a d d gi r, 2 0 0 7 ) VI. CONCLUSION Three coagulants shelled blended powder; deoiled powder and protein powder of the Moringa oleifera coagulant were used in the present study. Among them, protein powder has maximum removal efficiency compared to deoiled and shelled blended powder. The Shelled blended powder contains carbohydrates, oil and protein. The protein acts as a coagulant. The oil and carbohydrates present increase the organic matter contained in the treated water. This makes water unsuitable for drinking after storing it for some time. The deoiled powder contains protein and carbohydrates. The removal 20
  • 8. Comparative Study of Different Forms of Moringa Oleifera Extracts for Turbidity Removal of oil reduces the organic matter content of the water thus making water more suitable for drinking. The purified proteins reduce the organic matter content to the least. The reduction in the optimum dose required for shelled blended powder to deoiled powder to protein powder is because of the removal of oil and the carbohydrates, as only protein acts as a coagulant. In many villages of India, hundreds of acres of barren land are available. These lands can be utilized for cultivation of Moringa plants. As this plant does not require any specific soil type to grow, it can grow even in semi-arid regions as well. This plantation ultimately reduces the cost of chemical coagulants and also different byproducts of Moringa like leaf powder, Moringa oil and flowers are available. It can be of better economic benefits to locals. VII. ACKNOWLEDGEMENT The authors are grateful to Bharati Vidyapeeth Institute of Environment Education & Research Pune, and B.V.D.U. College of Engineering, Pune and Rajiv Gandhi Institute of I.T. & B.T. Pune. REFERENCES [1]. Ndabigengesere Anselme, Narasiah K.S Quality of water treated by coagulation using Moringa Oleifera seeds. Water Res. 32(3), 781-791(1998) [2]. Kawamura S. Effectiveness of natural polyelectrolytes in water treatment. JAWWA, 83(10), 88-91 (1991) [3]. Lee S.H., Lee S. O., Jang K.L., Lee T. H.,Microbial flocculant from arcuadendron SP – 49. Biotechnol. Lett. 17(1), 95 – 100 (2001) [4]. Jahn S.A.A. Using Moringa seeds as coagulants in developing countries. Journal of American Water Works Association, 80:43– 50 (1988). [5]. Muyibi S.A. and Evison L.M Optimising Physical Parameters affecting Coagulation of Turbid water with Moringa Oleifera seeds. Water Res. 29 (12), 2689-2695 (1995) [6]. Olsen Annette Low technology water purification by Bentonite clay and Moringa oleifera seed flocculation as performed in Sudanese villages: Effects on Schistosoma mansoni cercariae. Water Res. 21 (5) 517-522(1987) [7]. Anhwange B.A., Ajibola V.O., Oniye S.J. Chemical studies of the seeds of Moringa oleifera (Lam) and Detarium microcarpum (Guill and Spurr) J. Of Biological Sciences 4(6) 711-715 (2004) [8]. Ghebremichael K.A, Gunaratna K.R, Henriksson H,Brumer H, Dalhammar G. A. Simple purification and activity assay of the coagulant protein from Moringa oleifera Seed. Water Research 39: 2338-2344 (2005). [9]. Geoff Folkard, John Sutherland, Rod Shaw .water clarification using Moringa oleifera seed as coagulant. Val-3, (2007) [10]. Gassenschmidt U, Jany K K, et al. Isolation and characterization of a flocculation protein from MO lam. BBA Biochem Biophys Acta 1243: 477-481 (1995) [ 1 1 ] . Amelia Pilar Rauter .Natural products in new millennium prospects and Indusry, page no-405-416, (2002). 21