Accepted Manuscript

Title: A Contemporary Review on Plant-Based Coagulants for

Applications in Water Treatment

Authors: Mussarat Saleem, Robert Thomas Bachmann

PII: S1226-086X(18)31496-5
DOI: https://doi.org/10.1016/j.jiec.2018.12.029
Reference: JIEC 4322

To appear in:

Received date: 19 March 2018

Revised date: 3 December 2018
Accepted date: 15 December 2018

Please cite this article as: Mussarat Saleem, Robert Thomas

Bachmann, A Contemporary Review on Plant-Based Coagulants for
Applications in Water Treatment, Journal of Industrial and Engineering
Chemistry https://doi.org/10.1016/j.jiec.2018.12.029

This is a PDF file of an unedited manuscript that has been accepted for publication.
As a service to our customers we are providing this early version of the manuscript.
The manuscript will undergo copyediting, typesetting, and review of the resulting proof
before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that
apply to the journal pertain.
Full Title: A Contemporary Review on Plant-Based Coagulants for Applications in Water

Treatment

Short Title: Plant-Based Coagulants for Potable Water Treatment: Contemporary Review

T
IP
Mussarat Saleem1* and Robert Thomas Bachmann2

R
SC
Malaysian Institute of Chemical and Bioengineering Technology, Universiti Kuala Lumpur,

78000 Alor Gajah, Malaysia. U

N
A
Emails: mussarat.micet@gmail.com1&bachmann@unikl.edu.my2
M

*Corresponding author
D
TE

Graphical abstract
EP
CC
A

1
 T
R IP
SC
U
N
Abstract (150 words max.)
A

Conventional coagulants, like aluminium sulphate and ferric chloride, are used in potable
M

water treatment and involve non-sustainable mining and transformation of raw materials
D

for their production with costly sludge disposal. Natural coagulants are mostly obtained
TE

from bacteria, fungi, animals and plants and are classified as polysaccharide, amino-

polysaccharide, poly-phenols and proteins- based substances. Plant-based coagulants

EP

extracted from Moringa oleifera, Strychnos potatorum Linn, Plantago ovate, Trigonella
CC

foenum graecum and Opuntia ficus indica are potential substitutes to chemicals mostly

based on bench-scale testing. These are organic polymers and polyelectrolytes that are
A

classified as cationic, anionic and non-ionic coagulants. This paper provides a historical and

contemporary review of plant-based coagulants, their notable milestones achieved,

chemistry involved, as well as bench, pilot and full scale trials, highlighting the effects of

2
plant-based coagulants on physico-bio-chemical properties of raw water. Commercialisation

constrains are also included and discussed.

Keywords (max. 5): Potable water treatment; plant-based coagulants; organic polymers;

polyelectrolytes; chemistry.

T
R IP
SC
1. Introduction

Due to increasingly stringent requirements in quality, reliability, economics and

U
sustainability, water clarification using conventional coagulants has become a formidable
N
challenge. Many shortcomings are identified for such chemicals in general (Bates et al. [1];
A
Kleinman et al. [2]; King & Noss [3]; Letterman & Pero [4]; Packham [5];Radford [6]; Van
M

Hullebusch et al. [7]), and for alum, in particular, such as;

D

 High carbon footprint associated with production of coagulant. According to He et al.

TE

[8], an estimated 35 % of a water treatment plant’s carbon footprint is due to use of

EP

coagulant alum.

 Detrimental human health effects linked with presence of residual alum in treated
CC

waters (Bondy & Campbell [9]; Martyn et al. [10]; Rogers & Simon [11]).
A

 Environmental pollution due to improper disposal of raw alum sludge (Muisa et al.

[12]; Kaggwa et al. [13]; Mortula et al. [14]; Gutierrez et al. [15]).

3
  Costly practices of sludge disposal as hazardous waste in sanitary landfills, or as

simple spread on land (Maiden et al. [16]; Boaventura et al. [17]; Evuti & Lawal [18]).

In Australia, for instance, alum sludge disposal cost was estimated to be $130 per ton

(Maiden et al. [16])

 Health and environmental impacts linked with bauxite mining activities (Abdullah et

T
al. [19]).

IP
Plant-based coagulants represent a renewable, non-hazardous, degradable, potentially

R
carbon-neutral option, and are receiving increased attention for replacing conventional

SC
coagulants (Figure 1).The term ‘plant-based coagulants’ refers to natural, water-soluble,

U
organic, ionic (cationic, anionic, or poly-ionic) and non-ionic polymers of diverse molecular
N
weights, derived from various plant components (Bulusu et al. [20]; Jahn & Dirar[21]; Diaz et
A
al. [22]; Marobhe et al. [23];Mahungu & Meyland[24];Abidin et al. [25]; Muthuraman &
M

Sasikala [26]; Fatombi et al. [27];Bodlund et al. [28]; Dezfooli et al. [29]). They acquire
D

somewhat random configurations in colloid- free aqueous environments, and restricted

TE

irreversible arrangement of loops, tails and trains onto adsorbed colloid particles in

solutions (Bolto & Gregory [30]). Plant-based coagulants help destabilise colloidal
EP

suspensions and form micro- and macro- flocs through charge neutralisation

(Ndabigengesere et al. [31]; Ghebremichael et al. [32]; Marobhe et al. [23]) and inter-
CC

particle bridging (Miller et al. [33]; Antov et al. [34]; Hameed et al. [35]). Macro-sized flocs
A

are readily removed by sedimentation while micro-flocs may be subjected to either

flocculant aid or filtration (Bratby, [36]).

Charge neutralisation refers to adsorption of charged coagulants onto oppositely charged

colloidal surfaces with subsequent neutralisation; whilst bridging refers to adsorption of

4
coagulant segments onto adjacent colloidal surfaces (in the presence of divalent metal ions

usually), thereby binding them together (Bolto & Gregory [30]). In bridging, both coagulant

and colloid carry same nature of charges (Bratby [36]). Some plant-based substances

however may act as flocculant aid by strengthening the flocs and increasing their

settleability (Awang & Aziz [37];Al-Hamadani et al. [38]; Bhole [39]). In addition to plant-

T
based coagulants, other sources containing natural coagulants include bacteria (Toeda &

IP
Kurane [40];Yokoi et al. [41];Kumar et al. [42];Xiong et al. [43]), fungi (Abdel-Fattah &

R
Saleh[44]; Farshad et al. [45]), algae (Fattom & Shilo[46]), and animals (Kawamura[47];

SC
Roussy et al. [48]; Wang et al. [49]; Fast & Gude [50]). Therefore, the active compounds of

natural coagulants can be polysaccharides (Kebaili et al. [51]; Shamsnejati et al. [52]; Miller

U
et al. [33]; Prasertsan et al. [53]; Suh et al. [54]; Kurane & Nohata[55]; Toeda & Kurane[40]),
N
aminopolysaccharides (Kawamura et al. [47]), poly-phenolic substances (Özacar &
A

Şengil[56]; Graham et al. [57]; Sánchez-Martín et al. [58]; Sánchez-Martín et al. [59]),
M

functional proteins (Gassenschmidt et al. [60]; Ndabigenesere et al. [61]; Ghebremichael et

D

al. [32]), proteolytic enzymes (Horne & Banks[62]) or glycoproteins (Santos et al. [63];
TE

Ferreira et al. [64]).

EP

Research on plant-based coagulants has been carried out so far in at least 38 countries

originating from Asia, America, Africa and Europe. Most publications are from Malaysia,
CC

India and Brazil (Figure S1, SI), which are emerging economies in a climate zone that favours

the year-round growth of plants. The frequency of scientific publications on plant-based

A

coagulants alone has seen an exponential increase over the past five decades,

demonstrating the growing interest in this field (Figure1). The findings reported in figure 1

5
are based on both journal articles and conference proceedings which were extracted from

various repositories.

Several reviews on natural coagulants have been published (Kristianto[65]; Nandini & Sheba

[66];Oladoja [67]; Oladoja [68];Choy et al. [69]; Kansal & Kumari [70]; Choy et al. [71];

Theodoro et al. [72]; Bichi[73]; Yin[74]; Yongabi[75]; Bolto & Gregory [30]),the salient

T
features of which are summarised in Table 1. Major areas of interest of published reviews

IP
include process efficacy and mechanism; and coagulant properties and their applications.

R
The aim of this paper is to present a historic and contemporary review on landmarks,

SC
comprehensive chemical and structural attributes, marketing constrains, and process

outcomes of plant-based coagulants to close or highlight the existing knowledge gaps in

U
N
these areas. It is also important to remember that before the onset of modern water
A
treatment, plant-based coagulants have been used by various civilisations and communities
M

for at least 4000 years. The reported ancient use of plant-based coagulant inspired a new

generation of scientists seeking to develop sustainable alternatives to current water

D

treatment practices.
TE

2. Plant-based coagulants
EP

Plant-based knowledge for nutrition and medicine is observed in history during the 9000
CC

B.C. Earliest records on coagulation with plant-based substances (Baker [76]), on the

contrary, dates back to 2000 B.C. This suggests previous settlements to use multiple
A

rudimentary water clarification methods. The water quality of ancient times are well

illustrated in Bachmann and Edyvean [77] where other methods (such as cisterns (Vitruvius,

100B.C. Book VIII, 6:14 [78]) and use of wine (1 Timonthy 5:23) and gold or silver vessels

6
(Chattopadhyay [79]) are also stated that were used to improve the quality of drinking

water.

Numerous plant-based polymers and polyelectrolytes have been studied and tested for use

in coagulation since the late 1970s. Majority of tested coagulants belong to the family

Fabaceae, and are extracted from seeds (Sciban et al. [80]; Menkiti et al. [81]; Marobhe et

T
al. [23]) and leaves (Obuzor et al. [82]; Aweng et al. [83]). The most widely investigated

IP
plant-based coagulant, on the other hand, is M. oleifera seed proteins (Baptista et al. [84];

R
Camacho et al. [85]; Dezfooli et al. [29]) that belongs to the family Moringaceae.

SC
In laboratory studies, it has been demonstrated that plant-based coagulants are ionic and

U
non-ionic in nature. These polymers and polyelectrolytes are tested directly as coagulants
N
(Adinolfi et al. [86]; Miller et al. [33]; Bodlund et al. [28]) as well as chemically modified to
A
produce effective coagulants (Beltrán-Heredia et al. [87]; Ziolkowska et al. [88]).
M

On the basis of their proven performances, plant-based coagulants are workable substitutes
D

to current chemicals as coagulants and flocculant aids (Ghebremichael et al. [32]; Santos et
TE

al. [63], Özacar & Şengil [56]). A classical example is M. oleifera seed extracts that can be

used in clarifying high turbid water at 75-250mgL-1 dosages or at 100mgL-1 with 40mgL-1
EP

alum coagulant (Sutherland et al. [89]; Muyibi & Alfugara [90]). The M. oleifera working
CC

doses are within the typical range of alum that is used in a water treatment plant (Alley

[91]).
A

Plants with active coagulant ingredients have a potential to be commercialised like current

industrial crops. They offer wide range of active ingredients that can be harvested and used

in a conventional water treatment plant. An estimated 6.06 metric tonnes of M. oleifera

7
seeds, for instance, is successfully harvested in a hectare land area in Western Australia

(Biswas & John [92]). Six constraints in commercialisation are realised and reviewed, starting

from restricted supply of coagulants to low acceptance by investors.

Plant-based coagulants are discussed as follows;

2.1. Historic use and preparations

T
In ancient times, the fertile lands within the Tigris–Euphrates river system (Mesopotamia)

IP
and Eastern Mediterranean Sea (Levant) contained wild varieties of crop plants (such as wild

R
barley and wheat) and offered clean freshwater supplies in the form of springs, lakes and

SC
streams. As early as 11,000 B.C. nomadic humans began to settle down and form numerous

U
towns using mud bricks. Such primitive towns were eroded in heavy downpours and hence
N
rebuilt from time to time, up until they were abandoned (Frazee [93]). It is conceivable that
A
the earliest forms of freshwater contaminations included clay-based surface runoffs that
M

started with such settlements and it reflects the earliest challenge of ancient people to

tackle water-related challenges. After the advent of farming in 9000 B.C., ancient humans
D
TE

gained substantial plant-based knowledge that spread to near-by countries (Egypt, the Indus

Valley, China and Europe) (Frazee [93]). It seems reasonable to assume that through trade
EP

relations, many small communities (e.g. African, Asian, Aegean and other Aborigines) in

relative close proximity to the major civilisations of Mesopotamia, Egypt, Mohenjo-Daro,

CC

China, and Greece assimilated and developed the knowledge of plants for nutrition and
A

medicine purpose. Use of fermented drinks like beers in ancient Mesopotamia (Sumerians in

particular) (Frazee [93]) and ancient Egypt (Bunson[94]), caffeinated drinks in ancient Africa

and China (Weinberg & Bealer[95]; Campbell [96]) as well as rose water in medieval Europe

(Mower [97]) depict alternative ways adapted by ancient people in overcoming potential

8
thirst. Fermented milk (Greek Tarkhana) using plant materials in ancient Greece (Hills &

Bryer [98]) shows prospects of plants in altering aqueous chemistry.

Use of plant-based coagulants in water clarification can be traced back to 2000 B.C. Ancient

Egyptian inscriptions of the Middle Kingdom era reveal the use of almonds (Prunus dulcis) as

a clarifying agent. Empty clay vessels were smeared with crushed P. dulcis biomass followed

T
by subsequent addition of raw water. Rousing the mixture by hand for some time and a

IP
three hour standing period produced clean water, that was decanted to smaller jars for

R
consumption(Baker[76]). Ancient Egyptians, like primitive settlers presumed, used stone

SC
rolling pins as pestles and flat surfaces as mortars for grinding of plant material (Frazee

U
[93]); and coarse siltware vessels of variable sizes for short-term storage and cooling of
N
drinking water (Frazee [93]; Nicholson & Shaw[99]). Evidences from literary compositions
A
like Sanskrit writing (Suśruta sũtra 45.13) and other Indian manuscripts (Iyer [100]) provide
M

further examples on the use of plants as coagulants in ancient and medieval India. Such

writings conclude Strychnos potatorum (referred to as clearing nuts), Luffa cylindrical and
D

Phyllantus emblica as principal purifying substances, whilst other plants such as Cyperus
TE

rofundus, Andropogan muricatus and Emblic myrobalan were reported to gave taste and
EP

fragrance to clear water.

CC

Evidence of plant-based water treatment can also be found in other continents. Peruvian

texts from the 16thand 17thcentury stated the use of powdered roasted grains of Zea mays in
A

water by sailors during their voyages (Jahn[101]). Chilean folklore texts from the 19th

century refer to water clarification with Opuntia fiscus indica mucilages (Beckman & Lorenzo

[102]). Alfred Edmund Brehm, during his 1847 trip to Sudan, observed the use of P. dulcis

and Faba fona as coagulating agents by nomadic tribes (Brehm[103]).

9
Modern literature reports that rural communities of Tanzania and Sudan (descendants from

ancient Africa) as well as Peru and Mexico (descendants from ancient Columbia) used a wide

range of plant-based materials to cause coagulation of turbid water (Amin [104]; Kirchmer

et al. [105]; Jahn [106]; Jahn & Dirar [107]; Marobhe et al. [108]). Examples of plants that

appear to provide a good source of coagulants include roots of Maerua pseudopetalosa;

T
rhizome of Cyperus rotundus; seeds of Adansonia digitata, Arachis hypogaea, Moringa

IP
oleifera, M. peregrine, Lablab niger, Lupinus termis, Parkinsonia aculeata, Phoenix

R
dactylifera, Trigonella foenum graecum,Vigna unguiculata, Vicia faba, Voandzeia

SC
subterranean; the bark of Balanites aegyptiaca; branches of Caparis deciduas and Tamarix

nilotica; and leaves of Bergia suffruticosa, Opuntia ficus-indica, O. tuna and Salix suhserrata.

U
The traditional ways of using such plant materials in coagulation varies, depending on the
N
nature of the raw material. Usually materials containing bodily fluids (mostly roots and
A

leaves) are pounded to expose fleshy biomass causing the release of active coagulant
M

ingredients. Harvested seed, on the other hand, are dried, deshelled and milled. Hence, to
D

use Maerua pseudopetalosa for coagulation, roots are either crushed or chopped, and
TE

added to turbid water (Marobhe et al. [108]). Moringa oleifera seeds are known to be

mostly applied as powder to turbid water. Powders are usually obtained from deshelled
EP

seeds, which are ground with a grinding stone. After introducing the powder to turbid
CC

water(contained in large burnt clay containers), stirring by means of large wooden spoons

or bare hands is performed for some time, and clarified water is decanted carefully and
A

stored in traditional clay jars (Marobhe et al. [108]; Jahn [101]). In Sudan use of crushed

powder placed in cloth bag with a thread attached for stirring has been reported (Jahn &

Dirar[107]). This technique treats coagulant powder in a manner similar to crushed tea

leaves of tea bags. In addition to the use of seed powder, a small amount of water was also

10
used to extract coagulant in a small bowl with stirring encouraged for 10 min, followed by

addition of the extract to the turbid water(Ali et al. [109]; Jahn[101]). Recent scientific

studies have confirmed that the use of a short extraction time enhances the coagulation

efficacy, owing to release of active ingredients (Jung et al. [110]). In traditional use of O.

ficus-indica, leaves are broken and mucilaginous saps allowed to drop into turbid water

T
(Kirchmer et al. [105]). It is noteworthy to highlight that M. oleifera, legumes and various

IP
cacti, used by our ancestors, are still the most investigated plant species in 21st century lab-

R
based coagulation experiments (Figure S2, SI).

SC
2.2. Research milestones

Milestones in the field of water clarifying plant-based materials include i) the successful

U
isolation and characterisation of cationic proteins from M. oleifera seeds (Gassenschmidt et
N
al. [60]; Ndabigengesere et al. [61]; Ghebremichael et al. [32]), ii) the development of
A
M

simplified purification protocols for M. oleifera coagulant (Ghebremichael et al. [32];

Sánchez-Martín et al. [111]; Dezfooli et al. [29]), iii) the derivatisation of starch (Bratskaya,
D

[112]; Ziólkowska et al. [88]) and tannin molecules (Reed and Finck[113]; Sánchez-Martı ́n &
TE

Beltrán-Heredia [114]) to synthesise effective coagulants, iv) the over-expression of 6.5 kDa
EP

cationic protein from M. oleifera in Escherichia coli (Broin et al. [115]; Pavankumar et al.

[116]), and v) the use of bioreactors for large scale production of recombinant coagulant
CC

proteins (Pavankumar et al. [116]). In the latter milestone, the production of M. oleifera

recombinant protein is scaled up using 7 litre bioreactors, with 42mg of coagulant protein
A

purified per litre of over-expressed E. coli cell cultures, using super-paramagnetic iron oxide

nano-particles (Pavankumar et al. [116]).

11
Since most of the 400,000 plant species currently known (BGCI [117]) have not been

screened for coagulation activity, there is still a lot of research required. Patwardhan and

Gautam [118] proposed two main approaches for bio-prospection of natural products; i) the

classical approach, that relies on phytochemical factors, serendipity, and random screening;

and ii) selection of plants based on ancient literature, traditional knowledge, and practices.

T
The latter approach has so far been adopted by most research teams (Jahn & Dirar [107];

IP
Bhole [39]; Gunaratna et al. [119]; Mbogo[120]; Bodlund et al. [28]), confirming with

R
modern technology the coagulation activity of extracts from known plants (Table 2).In most

SC
cases, however, the actual compound(s) responsible for coagulation still remains unknown.

Bio-prospecting of natural coagulants may further be narrowed down by identifying the

U
order, the family and the part of plant that is most likely to contain target molecules. Most
N
plant-based coagulants are found in the seeds of Fabaceae (in the order of Fabales),
A

followed by the Malvaceae (order of Malvales), and various families under the order of
M

Sapindales (Figure S3, SI). From Table 3, seeds from the majority of plants are found to
D

constitute coagulant-active biomolecules. These seeds are relatively easy to harvest, handle
TE

and store, thereby making them convenient candidates for laboratory experimentations.
EP

In order to discover novel plant-based coagulants, modern approaches such as the

randomised screening (Patwardhan & Gautam [118]) should be adopted. Generally, four
CC

stages are distinguished in a randomised screen; pre-screens, screens, monitor and

secondary testings (Suffness & Pezzuto[121]). The most popular method used in laboratory
A

research with plant-based coagulants is jar testing. With the help of a simple apparatus and

stirring device, the coagulation process of a water treatment plant is simulated in a 1-2 litre

jar (Engelhardt [122]). However, jar tests are time and material intensive techniques

12
characterised primarily by their low throughputs, comparatively large quantities of reactants

required and waste produced. This makes the jar test a less convenient approach for

identifying and quantifying coagulation activities of a large number of plant samples for

randomised screens. In light of these challenges, assays involving 4 mL cuvettes coupled

with manual micropipettes (Ghebremichael et al. [32]), conical flasks coupled with rotary

T
shaker (Kurane & Nohata[123]; Okaiyeto et al. [124]), and optical microscopy (Broin et al.

IP
[115]) were developed. The cuvette-based approach, in particular, is increasingly

R
applied(Marobhe et al. [23], Santos et al. [63] and Ferreira et al. [64]). Advances in the use

SC
of micro-titre plate (MTP) based coagulation assays are reported to further reduce the

amount of coagulants and other reagents (Nordmark et al. [125]; Dezfooli et al. [29]). This is

U
equivalent to purified fractions obtainable from separation technologies such as HPLC and
N
SDS page. Use of MTPs to study coagulation activity of test samples offers advantages like
A

i)downscaling from 4 mL to 250 µL (Dezfooli et al. [29]) sample volumes; ii) high throughputs
M

(Nordmark et al. [125]; Dezfooli et al. [29]); and iii) availability of multiple mixing options for
D

inducing coagulation, like shaking (Nordmark et al. [125]; Dezfooli et al. [29]) or pipetting
TE

(Saleem et al. [126]). In addition, time and cost per sample is also further reduced, because

of automated monitoring systems in MTP readers, and the involvements of lesser quantities
EP

of materials.
CC

2.3. Chemistry

Once plant extracts with coagulation activity have been identified, it is important to purify,
A

identify and characterise the compound(s) responsible for coagulation. Based on our

understanding of coagulant chemistry in general (Bratby[36]), and known plant coagulants

in particular(Bolto & Gregory[30]), plant-based coagulants are classified into four types, i.e.

13
cationic, anionic, poly-ionic (ampholytes or amphoteros), and non- ionic (neutral)

coagulants.

At present, approximately 26% of active compounds have been isolated and identified from

various plant extracts (Table 2 and 3) calling for more concerted effort to close this

knowledge gap about plant extracts. Keeping in mind such limitations, this section reviews

T
the structure, reactivity and properties of reported plant based coagulants and also propose

IP
the identification of active compounds in uncharacterised plant extracts with coagulation

R
activity whenever possible.

SC
2.3.1. Cationic coagulants

U
Cationic coagulants are referred to as polymeric molecules that possess net positive
N
charges, typically at investigated coagulation pH of 7 or less (O’Brien & Novak[127]).
A
Examples include industrial cationic starches (Ziolkowska et al. [88]), seed proteins from M.
M

oleifera (Ndabigengesere et al. [61]; Ghebremichael et al. [32]), Cocos nucifera (Fatombi et
D

al. [128]), and Brassica sp. (Bodlund et al. [28]). Seed proteins from Vigna unguiculata
TE

(Marobhe et al. [23]) and Parkinsonia aculeate (Marobhe et al. [23]) can be categorised as

cationic, based upon the cationic medium used in chromatographic separation.

EP

2.3.1.1. Industrial cationic starches

CC

Industrial cationic starches, sometimes referred to as starch derivatives, are synthesized by

modifying starch with N-(3-chloro-2-hydroxypropyl) trimethylammonium chloride

A

(Harrington & Engelhardt[129]). Cationic functional groups such as CHR`N+ (CH3)3 are

attached to -OH group of starch molecule via ether linkage (-O-) (Harrington & Engelhardt

[129]). Industrial cationic starches are generally of medium molecular weight and medium

charge density (Bolto & Gregory[30]). The structure of an industrial cationic starch is shown

14
in Figure2. Reactivity of such coagulants involves the tertiary amine group. Since non-

bonded pair of electrons (NBEs) on nitrogen is shared with the fourth alkyl group, resultant

molecule has a good basicity. The degree of reactivity of industrial cationic starches depends

on the degree of cationic group ((CHR`N+ (CH3)3)) substitutions. A well-known example of

industrial cationic starch includes potato starch BORCET SZ 2000. This coagulant is

T
characterised as a highly substituted cationic coagulant with substitution degree values (SD)

IP
of 0.18-0.20. Since BORCET SZ 2000 has a higher degree of tertiary amine substituents, the

R
coagulant has lower water solubility (20%) (Ziolkowska et al. [88]).

SC
2.3.1.2. M. oleifera seed proteins

Seeds of M. oleifera are a rich source of cationic functional proteins reported as low
U
N
molecular weight cationic proteins designated as MO2X or MOCP (Gassenschmidt et al. [60];
A
Ndabigengesere et al. [61]; Ghebremichael et al. [32]; Shebek et al. [130]) or
M

hemagglutinating proteins called lectins (designated as cMoL) (Luz et al. [131]). Such

protein-based coagulants are commonly obtained through aqueous extraction of crushed

D

defatted or raw seeds (Ndabigengesere et al. [31]; Okuda et al. [132]; Ghebremichael et al.
TE

[133]; Santos et al. [63]; Sánchez-Martín et al. [134]) and exhibit impressive coagulation
EP

activity either individually (Broin et al. [135]) or in combination with multiple proteins

(Ghebremichael et al. [133]). Aqueous extracts are routinely purified based upon differences
CC

in solubility (e.g. ammonium sulphate precipitation) (Santos et al. [63]; Dezfooli et al. [29]),

charge (ion exchange chromatography) (Ghebremichael et al. [32]; Marobhe et al. [23];
A

Nordmark et al. [125]), size (SDS-PAGE) (Bodlund et al. [28]; Dezfooli et al. [29]), specific

binding (affinity chromatography) (Santos et al. [63]), hydrophobicity (reverse phase

chromatography) (Luz et al. [131]) and heat stability (Dezfooli et al. [29]) to obtain suitable

15
protein fractions. Gassenschmidt et al. [60] obtained an isolate designated as MO2.1 which is

often taken as a reference molecule for comparing other isoforms of MO2x, MOCP or cMoL

by other research groups.

2.3.1.2.1. MO2X proteins

MO2X proteins represent M. oleifera coagulant proteins, MO2.1 and MO2.2, which are

T
considered natural variants of ~6.5kDa peptide of 60 amino acid residues (Gassenschmidt et

IP
al. [60]; Ndabigengesere et al. [61]). The 6.5 kDa monomers are found to associate into a

R
dimers of ~ 13 kDa via disulfide bonds (Ndabigengesere et al. [61]) due to distinct presence

SC
of Cys residues located at positions - 12 and -25 (Gassenschmidt et al. [60]). Shebek et al.

[130], in addition, reported MO2.1 and MO2.2 proteins to even exist as tetramers of molecular
U
N
weight ~26 kDa. The MO2X proteins are also referred to as 2S albumin proteins, as they
A
share close structural homology to parts of napin B, a 2S albumin protein (Suarez et al.
M

[136]). Ghebremichael et al. [32] identified a mixture of peptides of molecular weight less

than 6.5 kDa in M. oleifera seed coagulant. All 5 peptides of molecular weights in between
D

1.3-2.13 kDa showed similarities with parts of MO2.1 sequence (SwissProt P24303).
TE

The structure of MO2.1coagulant predicted by using SSpro8 predictor is shown in Figure3.

EP

SSpro8 prediction results are, in fact, in conformity with Suarez et al. [136]. As apparent
CC

MO2X constitute of α-helices in 3 regions. Reactivity of MO2.1 coagulant involves positively

charged (7 Arg and 1 His), and polar amino acids (15Gln,4 Ser, 2Cys, 2Asn, 2Thr and 1Tyr),
A

respectively. Since MO2X constitutes of large fractions of Arg and His (11.7% Arg moles and

1.67%His moles) compared to Asp (1.67% moles), the isoelectric point (pI) reported for

MO2X in literature is >10 (Gassenschmidt et al. [60]; Ndabigengesere et al. [61]), thus

rendering MO2X a basic coagulant. MO2X were also found to be thermally stable proteins

16
(Ghebremichael et al. [32]). This behaviour of thermal stability can be used as a means of

purifying MO coagulant protein as deployed by Dezfooli et al. [29]who obtained a protein

purity of 94% at 121°C. Dezfooli et al. [29] for the first time classified MO coagulant polymer

as an intrinsically disordered protein (IDP) also known as “natively unfolded protein” based

on findings from computational tools.

T
2.3.1.2.2. cMoL proteins

IP
cMoL proteins represent M. oleifera lectins ( Luz et al. [131]). cMoL coagulant is a trimer of

R
~36 kDa molecular weight, has 101 amino acids, and is characterised as a heat-stable and pH

SC
resistant protein (Luz et al. [131]). Alignment of cMoL sequences 3 to 57, with MO2.1

sequences 5 to 59 shows distinct similarity between both the molecules (Figure 4). With

U
regard to structural details, cMoL contains 46% α-helix, 12% β-sheets, 17% β-turns and 25%
N
A
unordered structures (Luz et al. [131]).Due to the presence of positively charged amino
M

acids (16 Arg and 2 His), the computed pI found for cMoL coagulant is 11.7 (ExPASy), making

cMoL a basic coagulant. Like MO2X, reactivity of cMoL involves both positively charged and
D

polar amino acids.

TE

2.3.1.3. Cocos nucifera seed protein

EP

Seeds of C. nucifera contain a protein based coagulant called casein (Fatombi et al. [128]).

Purified casein, designated as CaSMG, is found to comprise of 5.6 kDa amino residues with
CC

an isoelectric point and charge density of 7.5 and 1.09 meq/g (Fatombi et al. [128]),
A

respectively. The presence of characteristic infra-red (IR) spectral peaks at 1649 cm-1, 1536

cm-1, and 1237 cm-1 suggesting CaSMG to contain a rich source of primary, secondary and

tertiary amide reactive groups (Fatombi et al. [128]). In addition, CaSMG is also found to

contain active -OH and -NH sites within its molecule due to the distinct presence of IR peaks

17
at 3411 cm-1 and 3290 cm-1, respectively (Fatombi et al. [128]). Due to dominance of

cationic amide functional groups in general, CaSMG is treated as a cationic coagulant.

2.3.1.4. Brassica sp. seed protein

Seeds of Brassica sp. of mustard large varieties (designated as MusL) were found to contain

a protein-based cationic coagulant (Bodlund et al. [28]). The MusL coagulant comprises of

T
19 amino acid residues, has a molecular weight of 6.5 kDa, is thermally stable (at 95°C), and

IP
has an amino acid sequence coverage similarity of 37 % with cationic M. oleifera, MO2.1

R
(SwissProt:P24303) (Bodlund et al. [28]). The structure of MusL coagulant, predicted using

SC
SSpro8, is shown in Figure 5. The results of SSpro8 predictor show MusL as a secondary

protein. The computed pI is 9.76 (ExPASy), hence making it a strong cationic coagulant.
U
N
Reactivity of MusL coagulant involves positively charged amino acids (1 Arg and 1 His), and
A
also polar amino acids (8Gln and 1Thr), respectively.
M

2.3.1.5. Vigna unguiculata and Parkinsonia aculeata seed proteins

D

Seeds of V. unguiculata and P. aculeate were found to constitute of proteins with

TE

coagulation activity (Marobhe et al. [23]). These coagulant proteins are indirectly

categorised as cationic due to involvement of CM sepharose matrix in obtaining pure

EP

fractions. The purified V. unguiculata and P. aculeate coagulants are found to share same

molecular mass of 6 kDa, exhibit remarkable heat tolerance and show significant
CC

coagulation activity at pH 5.5-8.5 (Marobhe et al. [23]), respectively.

A

2.3.2. Anionic coagulants

Anionic coagulants are referred to as polymeric molecules that possess net negative

charges, typically at investigated coagulation pH of 6.5- 8.5 or above (O’Brien &

Novak[127]). Examples include natural tannins (Özacar & Şengil [56]; Özacar & Şengil [137];

18
Sánchez-Martín et al. [58]), poly-galacturonic acid extracted from Opuntia mucilage (Miller

et al. [33]),and water-soluble seed proteins of Phaseolus vulgaris (Antov et al. [138]).

2.3.2.1. Natural tannins

Natural tannin with coagulant activity are commonly extracted from the bark or wood of

Acacia, Castanea and Schinopsis plants, as well as, the corn cup of Quercus macrolepis

T
(Özacar & Şengil[56]; Sánchez-Martín et al. [58]). Furthermore, coagulant extracts prepared

IP
from Castanea sativa and Mangifera indica seeds are proposed as anionic coagulant due to

R
the presence of natural tannins (Živković et al. [139]; Arogba[140]). Natural tannins are

SC
referred to as poly-phenolic compounds (Khanbabaee & Ree[141]), and classified as

condensed tannins (Kelly et al. [142]). Chemically, condensed tannins are a complex mixture
U
N
of poly-flavonoid with an average molecular weight of 1250 gmol-1 (Reed & Finck[113]). The
A
exemplary structure of natural tannin as a mono-flavonoid unit is shown in Figure6.
M

Reactivity of natural tannins involves phenolic -OH groups, present on resorcinol (labelled as

‘A’), and pyrogallol (labelled as ‘B’) rings. Because phenolic -OH groups exist as acidic or
D

phenolate anions due to resonance (Vermerris & Nicoholson[143]), natural tannins behave
TE

as anionic coagulants. Gaugler et al. [144] investigated the thermal stability of condensed
EP

tannins extracts from Radiata pine bark and established that pine bark extract fractions can

be processed at temperatures < 200°C if the co-extracted polysaccharides content is < 5

CC

%.Similar findings were reported by Lisperguer et al. [145] for Acacia bark suggesting that

tannins are even more heat stable than protein-based coagulants; thus, potentially widening
A

their applications to high-temperature processes in which an undesirable reaction by-

product is to be removed, for example. The pI of natural tannins from Acacia, Castanea and

Schinopsis plants has not been reported so far.

19
 2.3.2.2. Poly-galacturonic acid

Poly-galacturonic acid is a polysaccharide-based polymer that is an important constituent of

water-soluble extract of Opuntia mucilage (Miller et al. [33]). Poly-galacturonic acid is also

referred to as pectic acid (Aspinall & Cañas-Rodriguez[146]), or as poly 1, 4-α-D-

galacturonate (IUPAC). The structure of poly-galacturonic acid is shown in Figure 7.

T
Reactivity of poly-galacturonic acid involves –OH groups that represent acidic sites available

IP
for de-protonation.

R
2.3.2.3. P. vulgaris seed proteins

SC
Seeds of P. vulgaris constitute of an anionic protein-type coagulant fraction(Antov et al.

[138]). Chromatographic separation of the water-soluble extract of plant seeds using

U
N
AmberliteTM IRA 900 Cl was carried out by Antov et al. [138] that show the coagulant
A
fraction to comprise of predominately negatively charged amino acids. Although the studies
M

on P. vulgaris are still ongoing, the reactivity of this coagulant can be attributed due to

presence of negatively charged and/or polar amino acids.

D
TE

2.3.3. Poly-ionic coagulants

Poly-ionic coagulants are substances that act as either ampholytic (neutral) coagulants when
EP

used at coagulation pH of 7, or as amphoteric coagulants when used at acidic and basic pHs.
CC

Poly-ionic coagulants as neutrals are usually insoluble, whilst as amphoters are easily

dissolved in water. A well-known example of poly-ionic coagulant includes commercial

A

tannins (Reed & Finck[113]; Beltrán-Heredia et al. [87]). Commercial tannins, also referred

to as tannin Mannich polymers, are synthesized by modifying condensed tannins using

Mannich reactions (Reed & Finck[113]; Roux et al. [147]). The structure of commercial

tannin involving primary amine substitution is shown in Figure 8. Reactivity of commercial

20
tannins involves both cationic amines and anionic phenols. When used at investigated

coagulation pH of 7, the non-ionic character becomes dominant causing the solubility of

molecule to decrease and form flocs (Mitchell et al. [148]). However, when used at acidic

pH, positive charges become more accentuated due to protonation of amine groups, while

at basic pH, negative charges become more accentuated due to deprotonation of phenolic –

T
OH groups. For effective use in coagulation, commercial tannins must also contain some

IP
degree of polymerization within its structure; hence, primary amine substitutions are

R
preferred over secondary amines substitutions (Mitchell et al. [148]). Primary amines

SC
polymerise the tannins through methylene (-CH2-) bridges at active nucleophilic tannin sites

(which are 6- and 8- positions of the "A" resorcinol ring) (Mitchell et al. [148]). A list of

U
commercial tannins that are used in coagulation of turbid waters is provided in Table 4.
N
A
2.3.4. Non-ionic coagulants
M

Non-ionic coagulants are referred to as polymeric molecules that possess net zero charges,

typically at investigated coagulation pH of 6.5- 8.5 (O’Brien & Novak [127]). Examples of
D

non-ionic coagulants of plant origin include polysaccharide compounds like gums and native
TE

starches (Foster[149]; Levine[150]; Adinolfi et al. [151]; Otegui [152]; Kumar &
EP

Bhattacharya[153]; Choy et al. [154]). Native starches are complex polysaccharides

composed mainly of amylopectin molecules (Figure 9) (Schwartz & Whistler[155]). A well-

CC

known example of starch-based coagulant is Zea may starch (Mandloi et al. [156]; Schwartz

& Whistler[155]). Due to amylopectin composition, such coagulants are waxy in nature
A

(Schwartz & Whistler[155]), and adhesive towards turbid impurities. This following section

discusses gum-based natural coagulants in more detail.

21
 2.3.4.1. Plant gums

Polysaccharide-based plant gums are non-toxic and non-irritable hydrophilic polymers

composed of different mono-saccharides linked together through glycoside bonds (Kibbe

[157]). Aqueous solutions are highly viscous, pH stable (Kibbe [157]) and exhibit high

potential of synergistic interactions with other biomolecules due to active –OH sites.

T
Based on the nature of chemical constituents, plant gums are broadly classified into

IP
galactomannans (mostly heterogeneous); and non-galactomannans (Mirhosseini & Amid

R
[109]). Plant gums reported as non-ionic coagulants include Cyamopsis tetragonoloba and S.

SC
potatorum seed gums. Based on our understanding of gum chemistry (Mirhosseini & Amid

[109]) many plant gums (mostly originating from seeds, Table 3) are classified as non-ionic
U
N
coagulants. Such gums are safe to use as coagulants since they are biocompatible and
A
commonly used in foods and pharmaceutical formulations (Attama et al. [158];
M

Gowthamrajan et al. [159]; Ogaji [160]; Singh et al. [161]; Somboonpanyakul et al. [162];

Noorlaila et al. [163]). Examples of galactomannans include seed gums of C. tetragonoloba,

D

Ceratonia siliqu, Prosopis juliflora, Prosopis laevigata, T.foenum graecum and S. potatorum.
TE

Examples of non-galactomannans, on the other hand, include Hibiscus esculentus and O.

EP

fiscus indica pod gums and Dolichos lablab, Scaphium scaphigerum and Tamarindus indica

seed gums.
CC

2.3.4.1.1. Galactomannan- based gums

A

Galactomannans, which are also referred to as manno-galactan gums (Foster[149]),consist

of a mannose backbone with galactose side groups (Mathur[164]). Extracted gums with just

mannose and galactose constituents are categorised as homogeneous galactomannans,

whilst those with supplementary monosaccharide-based residues are called heterogeneous

22
or simply ‘complex galactomannans’. Gums are mostly isolated as complex galactomannans

rather than as homogeneous galactomannans.

Galactomannans, in general, are most abundant in legume seeds (Anderson[165]; Dea and

Morrison[166]). They differ from one another on the basis of mannose-to-galactose ratio

(Mathur[164]), and presence/ absence of other monosaccharide constituents (Mirhosseini &

T
Amid [109]). A simplified structure of a typical galactomannan is shown in Figure 10.

IP
Reactivity of manno-galactan polymers, in general, is due to –OH functional groups residing

R
on mannose, galactose and other neutral –ose (sugar) molecules. Such sites offer high

SC
capacity to establish H-bond networks with surrounding molecules.

2.3.4.1.1.1. Homogeneous galactomannans

U
N
2.3.4.1.1.1.1. T. foenum graecum seed gums
A
T. foenum graecum seed gums (or fenugreek gum) contain a 1.1:1 molar ratio of mannose-
M

to-galactose (Mathur[164]) that constitutes of 15% of total seed composition (Palanuvej et

D

al. [167]). The structure of T. foenum graecum seed gum offers 60 % of galactose associated
TE

-OH reactive sites, while the remaining 40% of -OH sites are mannose related. Fenugreek

gums are characterized as 32.3 kDa viscous polymers (Jiang et al. [168]).
EP

2.3.4.1.1.2. Complex galactomannans

CC

2.3.4.1.1.2.1. Cyamopsis tetragonoloba seed gums

Cyamopsis tetragonoloba seed gums (or guar gum) are large polymers (50-800kDa)
A

composed primarily of mannose and galactose following a 2:1 molar ratio (Mahungu &

Meyland [24]). Compared with galactose, mannose constituent offers 57% of –OH reactive

sites. Xylose and arabinose, on the other hand, are identified as supplementary residues

(Wade & Weller [169]).

23
 2.3.4.1.1.2.2. Ceratonia siliqua seed gums

Ceratonia siliqua seed gums (popularly known as carob bean gum or locust bean gum) are

found to contain a 4:1 molar ratio of mannose to galactose with an average molecular

weight of 5-8kDa (Haddarah et al. [170]). It is considered a complex galactomannan

primarily due to significant amounts of arabinose that is either co-precipitated with seed

T
gum or simply attached to galactose units as arabino-galactan (Kok [171]). Majority (73%) of

IP
-OH reactive groups are provided by mannose constituent.

R
2.3.4.1.1.2.3. Prosopis spp. seed gums

SC
Prosopis spp. seed gums (or mesquite seed gum) are complex branched galactomannans

with a molecular weight of 62kDa (Vernon-Carter et al. [172]). Mannose and galactose (in
U
N
the mole ratio of 2.8:1) form the major part of the gum composition, while the
A
supplementary residues arabinose, rhamnose, glucuronate and methyl-glucuronate are
M

found existing as individual monosaccharide or as simple oligosaccharide side chains

(Vernon-Carter & Sherman [173]).

D
TE

2.3.4.1.1.2.4. Strychnos potatorum seed gums

Strychnos potatorum seed gums were the first plant-based coagulant solution investigated
EP

at bench scale for treatment of surface turbid water (Bulusu et al. [174]) and found to
CC

constitute of 2.5 part mannose to 1 part galactose (Adinolfi et al. [86]). It is regarded as a

complex galactomannan due to presence of 1.7 part galactan polysaccharide with one part
A

galactomannan.

2.3.4.1.2. Non-galactomannan based gums

Non-galactomannan based gums are complex heterogeneous polysaccharide-based

polymers that lack presence of mannose monosaccharides. Major chemical constituents of

24
non-galactomannans are neutral sugars (usually arabinose, rhamnose, galactose and xylose

monosaccharides) and acidic monosaccharides (mainly uronic acids) linked together through

glycoside bonds (Mirhosseini & Amid [175]). Like manno-galactan polymers, reactivity of

non-galactomannans is due to –OH functional groups residing on neutral sugars. Such –OH

groups are involved in forming H-bond networks with surrounding molecules.

T
2.3.4.1.2.1. Dolichos lablab seed gums

IP
Seeds of D.lablab contain a rich source of water-soluble neutral non-galactomannans

R
SC
(Salimath & Tharanathan [176]). The gums contain galactose and arabinose in the mole ratio

of 2:1 and about 6 % of uronic acid contents (Salimath & Tharanathan [176]). Therefore, the

U
major constituent of gum is arabino-galactan (i.e. galactose and arabinose molecules linked
N
via glycoside bonds) with uronic acid as minor residue. Both neutral sugars are in equal
A
share of –OH reactive groups.
M

2.3.4.1.2.2. Hibiscus esculentus pod gums

D

Hibiscus esculentus pod gums (or okra gums) are classified as non-galactomannans and
TE

contain galactose, rhamnose and galacturonic acid as main constituents (in the mole ratio of
EP

1:1:1) (Zaharuddin et al. [177]). The relative–OH reactive groups in various constituents is of

order; galactose (50%)> galacturonic acid (33%)> rhamnose (17%).

CC

2.3.4.1.2.3. Opuntia ficus-indica mucilages

A

The cladodes (modified stems) of O. ficus-indica contain complex non-galactomannans

composed of galactose, xylose, arabinose, rhamnose residues, and galacturonic acid

respectively (McGarvie & Parolis [178]). The gum backbone contains galacturonic acid,

25
rhamnose and galactose; and xylose and arabinose are attached as side chains via glycoside

bonds (McGarvie & Parolis [178]).Galacturonic acid constitutes 23.4% of gum, where as

arabinose, rhamnose, xylose and galactose are found in the molar ratios of 1.0:1.7:2.5:4.1

(Matsuhiro et al. [179]).

2.3.4.1.2.4. Scaphium scaphigerum seed gums

T
IP
Scaphium scaphigerum seed gums (or malva nut gum) are complex non-galactomannans

composed of 31.9% arabinose, 29.2% galactose, 29.5% rhamnose, 6.4% uronic acid and

R
SC
small content of glucose, xylose and mannose (Somboonpanyakul et al. [162]). Hence the

backbone of malva nut gum is composed of galactose and arabinose with rhamnose as side

U
chains (in a mole ratio of 1.00:1.67:1.01) (Somboonpanyakul et al. [162]).
N
2.3.4.1.2.5. Tamarindus indica seed gums
A
M

Seeds of T. indica were found to constitute of neutral gummy polysaccharide (65%),

commonly referred to as TSP (Saettone et al. [180]). The chemical constituents of TSP are
D

neutral sugars (glucose, xylose and galactose), present in a molar ratio of 2.8:2.2:1.0 (Goyal
TE

et al. [181]). Reactivity of TSP involves –OH functional groups (43% by glucose; 29% each by
EP

xylose and galactose). TSP is characterised as a hydrophilic polymer with a molecular weight

of 700–880 kDa (Kaur et al. [182]).

CC

2.4. Process scale-up trials

A

Although research on plant-based coagulants has advanced tremendously, process

developments beyond lab-scale are still at a preliminary stage. Keeping in view process

26
development limitations, all 3 levels of testings, i.e. bench-, pilot-, semi-/ full- process scales

are reviewed in the following.

2.4.1. Bench scale coagulation

Bench scale coagulation testing of plant- based substances has shown to generate WHO

acceptable drinking waters (Table 5). Water-soluble coagulant extracts of plants exhibit

T
impressive coagulation capacity towards turbid particles and microbes, with some effects

IP
observed on physico-chemical parameters of water like alkalinity, hardness and COD (Table

R
5). Against total coliforms, plant extracts show a remarkable removal capacity of at least 96

SC
% (Sánchez Martín et al. [183];Ramamurthy et al. [184]). With respect to softening of hard

water, M. oleifera seed extracts were found effective, though at the expense of relatively
U
N
high dose applications (Muyibi & Evison[185]).
A
From bench-scale trials (exhibit Table 5), extracts of M. oleifera, S. potatorum, P. ovate and
M

T. faenum graecum seeds are proposed as suitable candidates for process-scale ups. On the
D

other hand, natural tannins and extracts of Corchorus tridens leaves and H. sabdariffa calyx
TE

are found to act as effective coagulant aids instead (Table 5).

Besides the benefits of low dosage and negligible effects on pH, plant extracts were also
EP

shown to generate less volumes of sludge with potential use as biosolid. For instance, M.
CC

oleifera seed extracts produce a minimum sludge volume of 1.5 mL/L, contrary to 7.5 mL/L

produced by alum under same dosage conditions (1mL/L) (Ndabigengesere &

A

Narasiah[186]). However, one setback identified includes the increase of COD in treated

waters if crude extracts are used (Sánchez Martín et al. [183]). An increase in COD is likely to

promote microbial growth while increasing the turnover of adsorbents used in the filtration

stage of potable water treatment. Using crude M. oleifera coagulant extracts in rural

27
communities to treat turbid water for instant consumption can be considered safe as no

detrimental effects have been found for Moringa seeds (McBurney et al. [187]). A high COD

might become an inconvenience if treated water is stored for longer durations, or is

intended for treatments with chlorine (Sánchez Martín et al. [183]). Adaptation of solar/UV

disinfection techniques in rural communities can resolve this issue (Dawney et al. [188]; Lea

T
[189]). However, formation of disinfection by-product should be investigated to confirm its

IP
safe use as drinking water.

R
2.4.2. Pilot scale coagulation

SC
Pilot-scale coagulation trials of plant-based substances in treatment plants are summarised

in Table 6. Results show practicality of M. oleifera seed extracts in reducing turbidity, total

U
coliforms, faecal coliforms and faecal streptococci counts. Coagulation treatments using
N
A
other plant substances proved effective after integrating necessary membrane- based
M

technology (Table 6).

D

2.4.3. Semi- and full process scale coagulation

TE

In literature, semi- and full process scale coagulation using plant-based coagulants was

found for M. oleifera seed extracts only to the best of our knowledge. Such coagulation
EP

trials of M. oleifera seed extracts were shown to be robust, comparatively cheap to install

and modular(Yongabi et al. [190];Sutherland et al. [89]). The outcomes of trials revealed M.
CC

oleifera as excellent source of removing both turbidity and microbial counts (Yongabi et al.
A

[190]) confirming findings from lab- and pilot-scale studies. In addition, full scale trial at

16m3/hr rate has achieved equivalent turbidity removal to alum (Sutherland et al.

[89]).However, to maintain continuous supply of water such treatment units require

abundant supply and large-scale processing of plants materials (Sutherland et al. [89]). For

28
convenience, however, Moringa extracts as pellets can also be used, and stored at room

temperature for at least 78 weeks as suggested by Garcia-Fayos et al. [191].

2.4.4. Organic loads- a limitation of plant-based substances in process scale-ups

As stated previously, the fundamental drawback of plant-based coagulants for their

application in process-scale-up is the increase of organic loads (as COD/BOD) in treated

T
water particularly if non-purified extracts are used. According to Baptista et al. [192], a

IP
direct correlation exists between the nature of coagulant extract prepared and the amounts

R
of organic loads in treated water. In literature, the most conventional method of preparing

SC
the coagulant extract is through the use of distilled water (DW) as solvent. Such extracts

show a high relative turbidity removal, for DW-M. oleifera extract in particular, 80-99
U
N
%turbidity removal was found for both surface water and synthetic water of high turbidity
A
(Muyibi &Evison[193]; Ndabigengesere et al. [61] Lea et al. [189]). The DW- M. oleifera
M

extract (1%) was found to contain approximately 630 mgL-1 of total proteins and 888 mgL-1

of DOC (Baptista et al. [192]). According to Baptista et al. [192], 0.06mL of such extract gives
D

an average 25.3% increase of DOC in treated waters. Contrary to this, concentrated DW

TE

extracts (0.04 mL) produces an average 7.6% increase of DOC in treated water (70% less
EP

than that of non- concentrated DW extract).

CC

The coagulation activity of plant-based coagulants, in general, are known to increase by

using salt solution as an extracting media in place of distilled water particularly when used
A

against low turbid water (Okuda et al. [194]; Baptista et al. [192]). This improvement in

coagulation is found due to the salting- in mechanisms of proteins, reflecting an increase in

the solubility of the active component upon increased ionic strength of a solution. The most

commonly used salt solution in making of the coagulant extract is sodium chloride (NaCl).

29
NaCl- M. oleifera extracts (1%) contain an approx. 4282 mgL-1 of total proteins and 1746

mgL-1 of DOC (Baptista et al. [192]). According to Baptista et al. [192], 1.29 mL of such

extract gives an average 25 % increase of DOC, it’s concentrated counterpart (at 0.57 mL)

does not add DOC in treated water. Hence use of NaCl- coagulant extract concentrates or

pellets can be proposed as a convincing solution to control coagulant assisted organic loads

T
in treated waters.

IP
Another promising solution to control coagulant assisted organic loads in treated waters is

R
production of purified coagulant from MO defatted biomass. MO seeds are found to contain

SC
about 4.4% coagulant polymer of high thermal tolerance (Dezfooli et al. [29]) which can be

proved useful in this respect.

U
N
2.5. Commercialisation constraints
A

The commercialisation of plant-based coagulants faces six major constraints. The first major
M

constraint is maintaining a steady supply of raw materials that meets the minimum quality
D

requirement. Some plants like M. oleifera produce mature, dried seeds only twice a year
TE

(Radovich [195]) and require processing and storage which can add to the cost. The second

challenge is to produce sufficient amounts to replace conventional coagulants. Data on

EP

aluminium sulphate consumption by the drinking water industry is difficult to obtain;

CC

however, the amount of drinking water produced is often reported in national statistics

publication. Assuming that 0.08 kg alum are required to treat 1 m 3 of surface water (Bonton
A

et al. [196]) it is then possible to estimate the quantity of alum that needs to be substituted.

For Malaysia as an example, the average volume of drinking water supplied per day in 2015

was reported to be 16,159 million litres, requiring an estimated 1,293 MT of alum per year.

In order to determine the amount of plant-based coagulant required to substitute alum

30
while maintaining similar treatment performance, an equivalent dosage ratio of 10:1 (w/w)

is assumed based on M. oleifera data (Valverde et al. [197]). The annual average yield of M.

oleifera seeds is reported to range between 3.03 to 6.06MT /ha (Biswas & John [92]) of

which 75 % represent dehulled M. oleifera seeds. Thus, the total M. oleifera plantation area

required in Malaysia to produce a minimum amount of 12,930 MT of seeds is approximately

T
49,257 ha. To put this into perspective this area is about 1 % of the area occupied by oil

IP
palm plantation in Malaysia(Kong et al. [198]). At present, the standing stock of M. oleifera

R
in commercial plantations worldwide is unknown. In India, M. oleifera trees are reported to

SC
be cultivated on an area of 61,600 ha (Boopathi [199]). If the fruits of current industrial

crops such as oil palm contain substances with coagulation activity, the near-future

U
substitution of alum is a real possibility in countries such as Malaysia. It should be
N
remembered however, that the seeds of M. oleifera themselves also contain 35 - 42 wt% oil
A

(Anwar & Bhanger[200]; Anwar & Rashid [201]), while other tree components can be
M

commercially utilised too. When grown on irrigated land, MO seeds are produced twice the
D

amount (6.06 MT/ha) compared to when harvested on the dryland (3.03 MT/ha) (Biswas &
TE

John [92]). It is noteworthy to mention that the land usage information for growing MO

intensively is directly correlated to tree to tree spacing for particular harvest. Generally for
EP

pod harvest, the recommended tree to tree spacing is 1.2 and 5 m between rows, so as to
CC

obtain the desirable population of 1666 trees/ha (Zaku et al. [202]).

The third challenge is related to the cost- effective preparation of coagulants. Use of crude
A

coagulant preparations that are prepared from pre-processed materials have been reported

to release unwanted carbon loads into treated waters, resulting in undesirable microbial

growth (Abaliwano et al. [203]; Jahn and Dirar[107]). Also the presence of such organic

31
impurities in treated waters can give rise to bio-fouling of treatment membranes and

distribution pipelines by encouraging microbial population (Wilbert[204]; LeChevallier et al.

[205]; Bachmann & Edyvean [77]). So far, the most reliable method for generating pure

coagulant preparations was provided by Ghebremichael et al. [206] and Sánchez-Martín et

al. [111]. However, use of ion-exchangers (specifically cation exchanger) is a cost-intensive

T
technology; putting constrains towards commercialisation of natural coagulants. Recently,

IP
an effective use of ammonium sulphate precipitation coupled with temperature treatment

R
proposed by Dezfooli et al. [29] illustrates a straightforward isolation of a sufficiently pure

SC
coagulant fraction. This holds a promising future in terms of effective coagulant

preparations at lower cost.

U
N
Lack of regulatory approvals on plant-based coagulants is another challenge faced in
A
commercialising this technology as toxicological studies are lacking for purified coagulant
M

fractions. Approval of 10 mg/L dosage of chitosan (an animal-based natural coagulant) for

use in potable water treatment (Kawamura[47]), can be taken as an encouraging sign to get
D

regulatory approvals for natural coagulants.

TE

Breakthrough in acceptability by potential investors for plant-based coagulants as a new

EP

product is another challenge. With economy constantly fluctuating, coupled with absence of

data on clear cut risks of such coagulant utilities, investing money in this new technology
CC

can be proven difficult. The only extensive economic analysis of plant based coagulants is
A

provided by Jahn[101] using M. oleifera as coagulant aid at Kanhan Water Works in Nagpur,

India. According to Jahn [101], for treating 1.74 million m3 water, a total of US$ 2370

(equivalent of 20,856 INR) was saved by reduction in alum use. This early study serves as an

example of practical-based scientific contributions towards commercialising natural

32
coagulants. Future cost evaluations should be carried out based on total cost assessment to

capture hidden costs such as hazardous sludge waste handling which will improve the

economical performance of natural coagulants substantially.

In light of these challenges, it thus becomes necessary to not only conduct significant

practical-based research, but also review the knowledge gaps of plant based coagulants

T
contributed by scientific communities (some of which are presented in Figure S4,SI). It is

IP
also critical that life cycle assessment studies (LCA) in this regard should be carried out to

R
evaluate socioeconomics impacts of plant-based coagulant utilities.

SC
3. Conclusions

U
Early historic recordings and literary texts of 2nd millennium contain information on plant-
N
based materials that were used to produce clean drinkable water. These sources suggest
A

Prunus dulcis, Strychnos potatorum, Luffa cylindrical, Phyllantus emblica and Optunia fiscus
M

indica to contain coagulant ingredients. Modern literature also identifies multiple valuable
D

plants that were applied by indigenous populations of Tanzania, Sudan, Peru and Mexico in
TE

water clarification. Nearly 43% of these have been verified through lab experiments and

showed positive coagulation activity. Microtitre plate based coagulation assays is a

EP

promising means for rapid multi-factorial screening of natural coagulants in near future.
CC

Reviewing of research publications show that most of the active ingredients are found in
A

plants belonging to Fabaceae. Overall, forty five (45) plants of diverse families are found to

contain coagulant biomolecules and nineteen (19) of them are successfully classified here

into cationic-, anionic-, polyionic- and non-ionic coagulants. The investigated coagulants are

33
popularly used in clarifying muddy water (both in crude and purified forms) at bench scale

levels.

Plant-based coagulants, mostly tested in extract forms, are excellent substitutes for

conventional chemicals and produce WHO acceptable drinking waters. Among them, M.

oleifera seed extracts, a representative of cationic coagulants, have been widely tested as

T
prime coagulant in pilot-, semi- and full- process scale tests demonstrating the technical

IP
feasibility to substitute alum. Natural coagulants are promising industrial crops whose

R
products are workable substitutes of chemicals. In Malaysia, for instance, an estimated

SC
49,257 hectare of M. oleifera plantation is required to produce the quantities of coagulant

U
that will show equivalent turbidity removal to alum. As industrial crops, plant-based
N
coagulants may also offer economic development activities for rural areas thereby
A
increasing GDP of countries.
M

4. Funding
D

This work was partially funded by grant 02-02-13-SF0014 from the Ministry of Science,
TE

Technology and Innovation of Malaysia (MOSTI).

EP

5. Acknowledgement
CC

We would like to thank Mr. Khalid Saleem for proof-reading the manuscript. The

constructive comments from two anonymous reviewers are gratefully acknowledged.

A

6. Declaration of interest

The authors declare there is no conflict of interest.

34
References

[1] A. L. Bates, W. H. Orem, J. W. Harvey, and E. C. Spiker, “Tracing sources of sulfur in the Florida
Everglades,” J. Environ. Qual., vol. 31, pp. 287–299, 2002.
[2] M. T. Kleinman et al., “Human exposure to ferric sulfate aerosol: Effects on pulmonary
function and respiratory symptoms,” Am. Ind. Hyg. Assoc. J., vol. 42, no. 4, pp. 298–304,
1981.
[3] D. J. King and R. R. Noss, “Toxicity of Polyacrylamide and Acrylamide Monome,” Rev. Environ.
Health, vol. 8, no. 1–4, 1989.
[4] R. D. Letterman and R. W. Pero, “Contaminants in polyelectrolytes used in water treatment,”
J. Am. Water Work. Assoc., vol. 82, no. 11, pp. 87–97, 1990.

T
[5] R. F. Packham, “Polyelectrolytes in water clarification,” Proc. Soc.Water Treat. Exam., vol. 16,
pp. 88–111, 1967.

IP
[6] N. P. Radford, Ecotoxocological impact of ferric sulphate on chironomid cultures and
profundal reservoir communities, 1st ed. National Rivers Authority, 1994.
[7] E. Van Hullebusch, V. Deluchat, P. M. Chazal, and M. Baudu, “Environmental impact of two

R
successive chemical treatments in a small shallow eutrophied lake: Part I. Case of aluminium

SC
sulphate,” Env. Pollut, vol. 120, no. 3, pp. 617–26, 2002.
[8] C. He, Z. Liu, and M. Hodgins, “Using life cycle assessment for quantifying embedded water
and energy in a water treatment system,” 2013.
[9] S. C. Bondy and A. Campbell, “Aluminum and Neurodegenerative Diseases,” in Advances in

U
Neurotoxicology: Environmental Factors in Neurodegenerative Diseases, vol. 1, M. Aschner
and L. G. Costa, Eds. Elsevier, 2017, pp. 131–156.
N
[10] C. N. Martyn, D. J. Barker, C. Osmond, E. C. Harris, J. A. Edwardson, and R. F. Lacey,
“Geographical relation between Alzheimer’s disease and aluminum in drinking water,”
A
Lancet, vol. 1, pp. 59–62, 1989.
[11] M. A. Rogers and D. G. Simon, “A preliminary study of dietary aluminium intake and risk of
M

Alzheimer’s disease,” Age Ageing, vol. 28, pp. 205 –209, 1999.
[12] N. Muisa, Z. Hoko, and P. Chifamba, “Impacts of alum residues from Morton Jaffray Water
Works on water quality and fish, Harare, Zimbabwe,” Phys. Chem. Earth, Parts A/B/C, vol. 36,
D

no. 14–15, pp. 853–864, 2011.

[13] R. C. Kaggwa, C. I. Mulaleto, P. Denny, and T. O. Okurut, “The impact of alum discharges on a
TE

natural tropical wetland in uganda,” Water Res., vol. 35, no. 3, pp. 795–807, 2001.
[14] M. Mortula, S. M. Bard, M. E. Walsh, and G. A. Gagnon, “Aluminum toxicity and ecological risk
assessment of dried alum residual into surface water disposal 1,” vol. 136, pp. 127–136, 2009.
[15] R. Gutierrez, S. Delvasto, and M. E., “Problem of alum sludge disposal in Cali, Colombia,” in
EP

14th International conference on solid waste technology and management, 1998, pp. 1–4.
[16] P. Maiden, M. T. W. Hearn, R. I. Boysen, P. Chier, M. Warnecke, and W. R. Jackson, “Alum
sludge re-use investigation (10OS-42),” 2015.
CC

[17] R. A. R. Boaventura, A. A. L. S. Duarte, and M. F. Almeida, “Aluminum recovery from water

treatment sludges.,” in Proceedings of International conference on water supply and water
quality, 2000.
[18] A. M. Evuti and M. Lawal, “Recovery of coagulants from water works sludge : A review,” Adv.
A

Appl. Sci. Res., vol. 2, no. 6, pp. 410–417, 2011.

[19] N. H. Abdullah, N. Mohamed, L. H. Sulaiman, T. A. Zakaria, and D. A. Rahim, “Potential Health
Impacts of Bauxite Mining in Kuantan,” Malaysian J. Med. Sci., vol. 23, no. 3, pp. 1–8, 2016.
[20] K. R. Bulusu, V. P. Thergaonker, D. N. Kulkarni, and B. N. Pathak, “Natural poly-electrolytes as
coagulant aid,” Environ. Heal., vol. 10, pp. 239–264, 1968.
[21] S. A. . Jahn and H. Dirar, “Studies on natural water coagulants in the Sudan with special
reference to Moringa oleifera seeds,” Water SA, vol. 5, pp. 90–97, 1979.
[22] A. Diaz, N. Rincon, A. Escorihuela, N. Fernandez, E. Chacin, and C. F. Forster, “A preliminary

35
 evaluation of turbidity removal by natural coagulants indigenous to Venezuela,” Process
Biochem., vol. 35, pp. 391–395, 1999.
[23] N. J. Marobhe, G. Dalhammar, and K. R. Gunaratna, “Simple and rapid methods for
purification and characterization of active coagulants from the seeds of Vigna unguiculata
and Parkinsonia aculeata,” Environ. Technol., vol. 28, no. 6, pp. 671–681, 2007.
[24] S. M. Mahungu and I. Meyland, “Cassia gum, chemical and technical assessment (CTA),”
2008.
[25] Z. Z. Abidin, N. S. Mohd Shamsudin, N. Madehi, and S. Sobri, “Optimisation of a method to
extract the active coagulant agent from Jatropha curcas seeds for use in turbidity removal,”
Ind. Crops Prod., vol. 41, pp. 319–323, Jan. 2013.
[26] G. Muthuraman and S. Sasikala, “Removal of turbidity from drinking water using natural

T
coagulants,” J. Ind. Eng. Chem., vol. 20, pp. 1727–1731, 2014.
[27] J. K. Fatombi, B. Lartiges, T. Aminou, O. Barres, and C. Caillet, “A natural coagulant protein

IP
from copra (Cocos nucifera): Isolation, characterization, and potential for water purification,”
Sep. Purif. Technol., vol. 116, pp. 35–40, 2013.
[28] I. Bodlund, P. A. R., R. Chelliah, S. Kasi, K. Sankaran, and G. K. Rajarao, “Coagulant proteins

R
identified in Mustard: a potential water treatment agent,” Int. J. Environ. Sci. Technol., vol.
11, pp. 873–880, 2014.

SC
[29] S. M. Dezfooli, V. N. Uversky, M. Saleem, F. S. Baharudin, S. M. S. Hitam, and R. T. Bachmann,
“A simplified method for the purification of an intrinsically disordered coagulant protein from
defatted Moringa oleifera seeds,” Process Biochem., vol. 51, no. 8, 2016.
[30]
U
B. Bolto and J. Gregory, “Review: Organic polyelectrolytes in water treatments,” Water Res.,
no. 41, pp. 2301–2324, 2007.
N
[31] A. Ndabigengesere, K. S. Narasiah, and G. B. Talbot, “Active agents and mechanism of
coagulation of turbid waters using Moringa oleifera,” Water Res., vol. 29, no. 2, pp. 703–710,
A
1995.
[32] K. A. Ghebremichael, K. . R. Gunaratna, H. Henriksson, H. Brumer, and G. Dalhammar, “A
M

simple purification and activity assay of the coagulant protein from Moringa oleifera seed,”
Water Res., vol. 39, pp. 2338–2344, 2005.
[33] S. M. Miller, E. J. Fugate, V. O. Craver, J. A. Smith, and J. B. Zimmerman, “Towards
D

understanding the efficacy and mechanism of Opuntia spp. as a natural coagulant for
potential application in water treatment,” Environ. Sci. Technol., vol. 42, pp. 4274–4279,
TE

2008.
[34] M. G. Antov, M. B. Śćiban, and N. J. Petrović, “Proteins from common beans (Phaseolus
vulgaris) seed as natural coagulant for potential application in water turbidity removal,”
Bioresour. Technol., vol. 101, pp. 2167–2172, 2010.
EP

[35] Y. T. Hameed, A. Idris, S. A. Hussain, and N. Abdullah, “A tannin-based agent for coagulation
and flocculation of municipal wastewater: Chemical composition, performance assessment
compared to Polyaluminum chloride, and application in a pilot plant,” J. Environ. Manage.,
CC

vol. 184 (Pt 3), pp. 494–503, 2016.

[36] J. Bratby, Coagulation and Flocculation in Water and Wastewater Treatment, Second. IWA
Publishing, 2006.
[37] N. A. Awang and H. A. Aziz, “Hibiscus rosa-sinensis leaf extract as coagulant aid in leachate
A

treatment,” Appl. Water Sci., vol. 2, pp. 293–298, 2012.

[38] Y. A. J. Al-Hamadani, M. S. Yusoff, M. Umar, M. J. K. Bashir, and M. N. Adlan, “Application of
psyllium husk as coagulant and coagulant aid in semi-aerobic landfill leachate treatment,” J.
Hazard. Mater., vol. 190, no. 1–3, pp. 582–587, 2011.
[39] A. G. Bhole, “Relative evaluation of a few natural coagulants,” J. Water Supply Res. Technol.,
vol. 44, no. 6, pp. 284–290, 1995.
[40] K. Toeda and R. Kurane, “Microbial flocculant from Alcaligenes cupidus KT201,” Agric. Biol.
Chem., vol. 55, pp. 2793–2799, 1991.

36
[41] H. Yokoi, T. Arima, J. Hirose, S. Hayashi, and Y. Takasaki, “Flocculating properties of poly (r-
glutamic acid) produced by Bacillus subtilis,” J. Ferment. Bioeng., vol. 82, no. 1, pp. 84–87,
1996.
[42] C. G. Kumar, H. S. Joo, J. S. Choi, Y. M. Koo, and C. S. Chang, “Purification and characterization
of an extracellular polysaccharide from hal- oalkalophilic Bacillus sp. I-450,” Enzyme Microb.
Technol., vol. 34, pp. 673–681, 2004.
[43] Y. Xiong et al., “Production and characterization of a novel bioflocculat from Bacillus
licheniformis,” Appl. Environ. Microbiol., vol. 76, no. 9, pp. 2778–2782, 2010.
[44] A. F. Abdel-Fattah and S. A. Saleh, “Production and isolation of milk-clotting enzyme from
Aspergillus versicolor,” Zbl. Bakt. II. Abt., vol. 134, pp. 547–550, 1979.
[45] K. Farshad, A. Soheila, and M. Fereydoon, “Semi-purification and kinetic study of microfungal

T
rennet biosynthesized by local isolate of Rhizomucor nainitalensis using solid-state
fermentation system: Concentration methods and determinant factors in clotting activity,”

IP
Eur. J. Exp. Biol., vol. 3, no. 2, pp. 167–174, 2013.
[46] A. Fattom and M. Shilo, “Phormidium J-1 bio-flocculant production and activity,” Arch.
Microbiol., vol. 139, pp. 421–426, 1984.

R
[47] S. Kawamura, “Effectiveness of natural polyelectrolytes in water treatment,” J. Am. Water
Work. Assoc., vol. 83, no. 10, pp. 88–91, 1991.

SC
[48] J. Roussy, M. Van Vooren, B. A. Dempsey, and E. Guibal, “Influence of chitosan characteristics
on the coagulation and the flocculation of bentonite suspensions,” Water Res., vol. 39, no.
14, pp. 3247–3258, 2005.
[49]
U
J.-P. Wang, Y.-Z. Chen, Y. Wang, S.-J. Yuan, G.-P. Sheng, and H.-Q. Yu, “A novel efficient
cationic flocculant prepared through grafting two monomers onto chitosan induced by
N
Gamma radiation,” RSC Adv., vol. 2, pp. 494–500, 2012.
[50] S. A. Fast and V. G. Gude, “Ultrasound-chitosan enhanced flocculation of low algal turbid
A
waters,” J. Ind. Eng. Chem., vol. 24, pp. 153–160, 2015.
[51] M. Kebaili, S. Djellali, M. Radjai, N. Drouiche, and H. Lounici, “Valorization of orange industry
M

residues to form a natural coagulant and adsorbent,” J. Ind. Eng. Chem., vol. 64, pp. 292–299,
2018.
[52] S. Shamsnejati, N. Chaibakhsh, A. R. Pendashteh, and S. Hayeripour, “Mucilaginous seed of
D

Ocimum basilicum as a natural coagulant for textile wastewater treatment,” Ind. Crops Prod.,
vol. 69, pp. 40–47, 2015.
TE

[53] P. Prasertsan, W. Dermlim, H. Doelle, and J. F. Kennedy, “Screening, characterization and

flocculating property of carbohydrate polymer from newly isolated Enterobacter cloacae
WD7,” Carbohydr. Polym., vol. 66, pp. 289–297, 2006.
[54] H. H. Suh, G. S. Kwon, C. H. Lee, S. H. Kim, H. M. Oh, and B. D. Yoon, “Characterization of
EP

bioflocculant produced by Bacillus sp. DP-152,” J. Ferment. Bioeng., vol. 84, no. 2, pp. 108–
112, 1997.
[55] R. Kurane and Y. Nohata, “Microbial flocculation of waste liquids and oil emulsions by a
CC

bioflocculant from Alcaligenes latus,” Agric. Biol. Chem., vol. 55, no. 4, pp. 1127–1129, 1991.
[56] M. Özacar and I. A. Şengil, “The use of tannins from Turkish acorns (valonia) in water
treatment as a coagulant and coagulant aid,” Turkish J. Eng. Environ. Sci., vol. 26, no. 3, pp.
255–263, 2002.
A

[57] G. N., G. F., F. J., and M. Watts, “Characterisation and coagulation performance of a tannin-
based cationic polymer: a preliminary assessment,” Colloids Surfaces A, vol. 327, pp. 9–16,
2008.
[58] J. Sánchez-Martín, M. González-Velasco, and J. Beltrán-Heredia, “Surface water treatment
with tannin-based coagulants from Quebracho (Schinopsis balansae),” Chem. Eng. J., vol. 165,
no. 3, pp. 851–858, 2009.
[59] J. Sánchez-Martín, J. Beltrán-Heredia, and C. Solera-Hernández, “Surface water and
wastewater treatment using a new tannin-based coagulant. Pilot plant trials.,” J. Environ.

37
 Manage., vol. 91, pp. 2051–2058, 2010.
[60] U. Gassenschmidt, K. D. Jany, B. Tauscher, and H. Niebergall, “Isolation and characterization
of a flocculating protein from Moringa oleifera Lam.,” Biochim. Biophys. Acta, vol. 1243, no. 3,
pp. 477–81, Apr. 1995.
[61] A. Ndabigengesere, K. S. S. Narasiah, and B. G. Talbot, “Active agents and mechanism of
coagulation of turbid waters using Moringa oleifera,” Water Res., vol. 29, no. 2, pp. 703–710,
Feb. 1995.
[62] D. S. Horne and J. M. Banks, Cheese:chemistry, physics and microbiology. New York, NY:
Elsevier Academic Press, 2003.
[63] A. F. S. Santos, L. A. Luz, A. C. C. Argolo, J. A. Teixeira, P. M. G. Paiva, and L. C. B. B. Coelho,
“Isolation of a seed coagulant Moringa oleifera lectin,” Process Biochem., vol. 44, pp. 504–

T
508, 2009.
[64] R. S. Ferreira et al., “Coagulant and anti-bacterial activities of the water-soluble seed lectin

IP
from Moringa oleifera,” Lett. Appl. Microbiol., vol. 53, pp. 186–192, 2011.
[65] H. Kristianto, “The Potency of Indonesia Native Plants as Natural Coagulant: a Mini Review,”
Water Conserv. Sci. Eng., vol. 2, pp. 51–60, 2017.

R
[66] G. K. M. Nandini and M. C. Sheba, “Emanating Trends in the Usage of Bio-coagulants in
Potable Water Treatment : a Review,” Int. Res. J. Eng. Technol., vol. 3, no. 11, pp. 970–974,

SC
2016.
[67] N. A. Oladoja, “Advances in the quest for substitute for synthetic organic polyelectrolytes as
coagulant aid in water and wastewater treatment operations,” Sustain. Chem. Pharm., vol. 3,

[68]
pp. 47–58, 2016.
U
N. A. Oladoja, “Headway on natural polymeric coagulants in water and wastewater treatment
N
operations,” J. Water Process Eng., vol. 6, pp. 174–192, 2015.
[69] Y. S. Choy, K. M. N. Prasad, T. Y. Wu, and R. N. Ramanan, “A review on common vegetables
A
and legumes as promising plant-based natural coagulants in water clarification,” Int. J.
Environ. Sci. Technol., vol. 12, pp. 367–390, 2015.
M

[70] S. K. Kansal and A. Kumari, “Potential of M. oleifera for the Treatment of Water and
Wastewater,” Chem. Rev., vol. 114, p. 4993−5010, 2014.
[71] Y. S. Choy, K. M. N. Prasad, T. Y. Wu, M. E. Raghunandan, and R. N. Ramanan, “Invited article:
D

Utilization of plant-based natural coagulants as future alternatives towards sustainable water

clarification,” J. Environ. Sci., vol. 26, pp. 2178 – 2189, 2014.
TE

[72] J. D. P. Theodoro, G. F. Lenz, R. F. Zara, and R. Bergamasco, “Coagulants and natural

polymers: Perspectives for the treatment of water,” Plast. Polym. Technol., vol. 2, no. 3, pp.
55–62, 2013.
[73] M. H. Bichi, “A Review of the applications of Moringa oleifera seeds extract in water
EP

treatment,” Civ. Environ. Res., vol. 3, pp. 1–9, 2013.

[74] C. Y. Yin, “Review: Emerging usage of plant-based coagulants for water and wastewater
treatment,” Process Biochem., vol. 45, pp. 1437–1444, 2010.
CC

[75] K. A. Yongabi, “Biocoagulants for Water and Waste Water Purification: a Review,” Int. Rev.
Chem. Eng., vol. 2, no. 3, pp. 444–458, 2010.
[76] M. N. Baker, The quest for pure water. New York, NY: American Water Works Association,
1949.
A

[77] R. T. Bachmann and R. G. J. Edyvean, “Biofouling: an historic and contemporary review of its
causes , consequences and control in drinking,” Biofilms, vol. 2, pp. 197–227, 2005.
[78] M. H. Morgan, “Vitruvius (100BC) De Architectura,” in The Ten Books on Architecture,
Cambridge: MA: Harvard University Press, 1914.
[79] A. Chattopadhyay, “Susruta recommends golden and jewelled veseels for eating and
drinking,” Indian J. Hist. Med., vol. 13, pp. 4–6, 1968.
[80] M. B. Śćiban, M. T. Klašnja, and Stojimirović., “Investigation of coagulation activity of natural
coagulants from seeds of different leguminose species,” APTEFF, vol. 36, pp. 81–87, 2005.

38
[81] M. C. Menkiti, P. C. Nnaji, C. I. Nwoye, and O. D. Onukwuli, “Coag-flocculation kinetics and
functional parameters response of Mcuna seed coagulant to pH variation in organic rich coal
effluent medium,” J. Miner. Mater. Charact. Eng., vol. 9, no. 2, pp. 89–103, 2010.
[82] G. U. Obuzor and M. Ejimozor, “Analysis and treatment of produced water (Igbo-etche) using
natural biocoagulant from Inga edulis,” Sci. Africana, vol. 10, no. 1, pp. 112–117, 2011.
[83] E. R. Aweng, A. I. Anwar, M. I. Siti Rafiqah, and O. Suhaimi, “Cassia alata as a potential
coagulant in water treatment,” Res. J. Recent Sci., vol. 1, no. 2, pp. 28–33, 2012.
[84] A. T. . Baptista, M. O. Silva, R. G. Gomes, R. Bergamasco, M. F. Vieira, and A. M. . Vieira,
“Protein fractionation of seeds of Moringa oleifera lam and its application in superficial water
treatment,” Sep. Purif. Technol., vol. 180, pp. 114–124, 2017.
[85] F. P. Camacho, V. S. Sousa, R. Bergamasco, and M. R. Teixeira, “The use of Moringa oleifera as

T
a natural coagulant in surface water treatment,” Chem. Eng. J., vol. 313, pp. 226–237, 2017.
[86] M. Adinolfi et al., “Composition of the coagulant polysaccharide fraction from Strychnos

IP
potatorum seeds,” Carbohydr. Res., vol. 263, pp. 103–110, 1994.
[87] J. Beltrán-Heredia, J. Sánchez-Martín, and M. C. Gόmez-Muńoz, “New coagulant agents from
tannin extracts: preliminary optimization studies,” Chem. Eng. J., vol. 162, no. 3, pp. 1019–

R
1025, 2010.
[88] D. ZiólKowska and A. Shyichuk, “Flocculation abilities of industrial cationic starches,”

SC
Polimery, vol. 56, no. nr3, pp. 244–246, 2011.
[89] J. P. Sutherland, G. K. Folkard, M. A. Mtawali, and W. D. Grant, “Moringa oleifera at pilot/full
scale. E-water treatment,” in water, sanitation, environment and development, Proceedings

[90] U
of 19th WEDC conference, Ghana: ACCRA, 1993, pp. 109–110.
S. A. Muyibi and A. M. S. Alfugara, “Treatment of surface water with Moringa oleifera seed
N
extract and alum - a comparative study using a pilot scale water treatment plant,” Int. J.
Environ. Stud., vol. 60, no. 6, pp. 617–626, Dec. 2003.
A
[91] E. R. Alley, “Appendix D: Chemical Characteristics,” in Water Quality Control Handbook,
McGraw Hill,Inc., 2000.
M

[92] W. K. Biswas and M. B. John, “Life cycle assessment of biodiesel production from moringa
oleifera oilseeds,” 2009.
[93] C. A. Frazee, World History: Ancient and medieval times to A.D. 1500, 1st ed. Barron’s
D

Educational Series, 1997.

[94] M. Bunson, Encyclopedia of Ancient Egypt, Revised Ed. Infobase Publishing, 2014.
TE

[95] B. A. Weinberg and B. K. Bealer, The World of Caffeine: The Science and Culture of the World’s
Most Popular Drug. New York, NY: Routledge, 2004.
[96] D. L. Campbell, The Tea Book. Pelican Publishing Company, 1995.
[97] R. M. Mower, “Rose water,” in The Oxford Encyclopedia of food and drink in America, Volume
EP

2., A. F. Smith, Ed. USA: Business & Economics, 2013.

[98] S. Hills and A. Bryer, “Byzantine Poridge: Tracta, trachanas & tarhana,” in Food in antiquity, J.
Wilkins, D. Harvey, and M. Dobson, Eds. Exeter University Press, 1995, pp. 44–54.
CC

[99] P. . Nicholson and I. Shaw, Ancient Egyptian Materials and Technology. Cambridge University
Press, 2000.
[100] N. C. Iyer, The Brhat samhita of Varaha mihira. Madura: Souts Indian Press, 1884.
[101] S.A.A Jahn, Traditional Water Purification in Tropical Developing Countries: Existing Methods
A

and Potential Application. Germany: German Agency for Technical Cooperation (GTZ), 1981.
[102] R. Beckman and W. Lorenzo, “Polyacrylamide water clarifier,” US20060180551 A1, 2006.
[103] A. E. Brehm, Reisen in Sudan. 1975.
[104] M. M. E. Amin, “A general list of common forest trees and shrubs of the Sudan.” Soba, 1973.
[105] C. J. Kirchmer, J. V. Arboleda, and M. L. Castro, “Natural polymers and their application as
flocculation assistants,” Lima, Peru, 1975.
[106] S. A. A. Jahn, “Traditional methods of water purification in the riverain Sudan in relation to
geographic and socioeconomic conditions,” Erdkunde, vol. 31, no. 2, pp. 120–130, 1977.

39
[107] S. A. A. Jahn and H. Dirar, “Studies on natural water coagulants in the Sudan, with special
reference to Moringa oleifera seeds,” Water South Africa, vol. 5, no. 2, pp. 90–104, 1979.
[108] N. J. Marobhe, G. Renman, and J. Gunnar, “The study of water supply and traditional water
purification knowledge in selected rural villages in Tanzania,” in Indigenous knowledge
systems and sustainable development: Relevance for Africa, Tribes and., E. Boon and L. Hens,
Eds. Delhi, Indian: Kamla Raj Enterprises, 2007, pp. 111–120.
[109] N. Ali, N. Mohammed, and A. Badri, “No Title,” in Women and the environment in the Sudan,
Environmen., D. Baxter, Ed. Institute of environmental studies, University of Khartoum, 1981.
[110] Y. Jung, Y. Jung, M. Kwon, H. Kye, Y. W. Abrha, and J. W. Kang, “Characteristic, Evaluation of
Moringa oleifera seed extract by extraction time: effect on coagulation efficiency and
extract,” Water Heal., 2018.

T
[111] J. Sánchez-Martín, K. Ghebremichael, and J. Beltrán-Heredia, “Comparison of single-step and
two-step purified coagulants from Moringa oleifera seed for turbidity and DOC removal,”

IP
Bioresour. Technol., vol. 101, pp. 6259–61, Aug. 2010.
[112] S. Bratskaya, S. Schwarz, T. Liebert, and T. Heinze, “Starch derivatives of high degree of
functionalization,” Colloids Surfaces A Physicochem. Eng. Asp., vol. 254, no. 1–3, pp. 75–80,

R
Mar. 2005.
[113] P. E. Reed and M. R. Finck, “Modified tannin Mannich polymers,” United State Patent No.

SC
5,659,002, 1997.
[114] J. Sanchez-Martıń and J. Beltran-Heredia, “Nature is the answer: water and wastewater
treatment by new natural-based agents,” in Advances in Water Treatment and Pollution

U
Prevention, S. K. Sharma and R. Sanghi, Eds. Springer Science & Business Media, 2012.
[115] M. Broin, C. Santaella, S. Cuine, K. Kokou, G. Peltier, and T. Joet, “Flocculent activity of a
N
recombinant protein from Moringa oleifera Lam seeds,” Appl Microbiol Biotechnol, vol. 60,
pp. 114–119, 2002.
A
[116] A. R. Pavankumar, J. Norén, L. Singh, and N. K. C. Gowda, “Scaling-up the production of
recombinant Moringa oleifera coagulant protein for large-scale water treatment
M

applications,” RSC Adv., vol. 4, pp. 7136–7141, 2013.

[117] Botanic Gardens Conservation International, “Plant species number,” Policy, 2010. [Online].
Available: https://www.bgci.org/policy/1521/.
D

[118] B. Patwardhan and M. Gautam, “Botanical immunomodulators and Chemoprotectants in

Cancer Therapy,” in Drug Discovery & Development, vol. 1, M. S. Chorghade, Ed. John Wiley &
TE

Sons, Inc., 2006, pp. 405–424.

[119] K. R. Gunaratna, B. Garcia, S. Andersson, and G. Dalhammar, “Screening and evaluation of
natural coagulants for water treatment,” Water Sci. Technol. Water Supply, vol. 7, no. 5–6, p.
19, Dec. 2007.
EP

[120] S. A. Mbogo, “A novel technology to improve drinking water quality using natural treatment
methods in rural Tanzania.,” J. Environ. Health, vol. 70, no. 7, pp. 46–50, Mar. 2008.
[121] M. Suffness and M. Pezzuto, “Assays related to cancer drug discovery,” in Method in Plant
CC

biochemistry, Vol. 6. As., K. Hostettmann, Ed. London: Academic Press, 1991, pp. 71–153.
[122] T. L. Engelhardt, Coagulation, Flocculation and Clarification of Drinking Water. HACH, 2010.
[123] R. Kurane and Y. Nohata, “A new water-absorbing polysaccharide from Alcaligenes latus,”
Biosci. Biotechnol. Biochem., vol. 58, no. 2, pp. 235–238, 1994.
A

[124] K. Okaiyeto, U. U. Nwodo, A. S. Okoli, L. V. Mabinya, and A. I. Okoh, “Studies on bioflocculant

production by Bacillus sp. AEMREG7,” Polish J. Environ. Stud., vol. 25, no. 1, pp. 241–250,
2016.
[125] B. A. Nordmark, T. M. Przybycien, and R. D. Tilton, “Comparative coagulation performance
study of Moringa oleifera cationic protein fractions with varying water hardness,” J. Environ.
Chem. Eng., vol. 4, pp. 4690–4698, 2016.
[126] M. Saleem, R. T. Bachmann, and M. Ahmad, “Isolation and characterization of water-soluble
partially purified polymer fractions of Moringa oleifera seeds and its flocculating activity using

40
 a novel microtitre plate-based assay,” in 13th National Symposium on Polymeric Materials
(NSPM), 2013.
[127] James H. O’Brien and J. T. Novak, “Effects of pH and Mixing on Polymer Conditioning of
Chemical Sludge,” J. Am. Water Works Assoc., vol. 69, no. 11, pp. 600–605, 1977.
[128] J. K. Fatombi, B. Lartiges, T. Aminou, O. Barres, and C. Caillet, “A natural coagulant protein
from copra (Cocos nucifera): Isolation, characterization, and potential for water purification,”
Sep. Purif. Technol., vol. 116, pp. 35–40, 2013.
[129] A. Harrington and P. Engelhardt, “Cationic starches as industrial flocculants,” in Proceedings
of the International Water Conference, 1979, pp. 313–316.
[130] K. Shebek, A. B. Schantz, I. Sines, K. Lauser, S. Velegol, and M. Kumar, “The flocculating
cationic polypetide from Moringa oleifera seeds damages bacterial cell membranes by

T
causing membrane fusion,” Langmuir, vol. 31, no. 15, pp. 4496–4502, 2015.
[131] L. D. A. Luz et al., “Structural characterization of coagulant Moringa oleifera Lectin and

IP
itseffect on hemostatic parameters,” Int. J. Biol. Macromol., vol. 58, pp. 31– 36, 2013.
[132] T. Okuda, A. U. Baes, and W. Nishijima, “Isolation and characterization of coagulant extracted
from Moringa oleifera seed by salt solution,” Wat. Res., vol. 35, no. 2, pp. 405–410, 2001.

R
[133] K. A. Ghebremichael, K. R. Gunaratna, H. Henriksson, H. Brumer, and G. Dalhammar, “A
simple purification assay of the coagulant protein from Moringa oleifera seed,” Water Res.,

SC
vol. 39, no. 11, pp. 2338–2344, 2005.
[134] J. Sanchez-Martin, K. Ghebremichael, and J. Beltrán-Heredia, “Comparison of single-step and
two-step purified coagulants from Moringa oleifera seed for turbidity and DOC removal,”

[135]
Bioresour. Technol., vol. 101, pp. 6259–6261, 2010.
U
M. Broin, C. Santarlla, S. Cuine, K. Kokou, G. Peltier, and T. Joet, “Flocculant activity of a
N
recombinant protein from Moringa oleifera Lam seeds.,” Appl. Microbiol. Biotechnol., vol. 60,
pp. 114–119, 2002.
A
[136] M. Suarez et al., “Structure-Function Characterization and Optimization of a Plant-Derived
Antibacterial Peptide,” vol. 49, no. 9, pp. 3847–3857, 2005.
M

[137] M. Özacar and İ. A. Şengil, “Evaluation of tannin biopolymer as a coagulant aid for coagulation
of colloidal particles,” Colloids Surfaces A Physicochem. Eng. Asp., vol. 229, no. 1–3, pp. 85–
96, Nov. 2003.
D

[138] M. G. Antov, M. B. Šćiban, and N. J. Petrović, “Proteins from common bean (Phaseolus
vulgaris) seed as a natural coagulant for potential application in water turbidity removal,”
TE

Bioresour. Technol., vol. 101, no. 7, pp. 2167–2172, 2010.

[139] J. Živković, I. Mujić, Z. Zeković, G. Nikolić, S. Vidović, and A. Mujić, “Extraction and analysis of
condensed tannins in Castanea Sativa Mill,” J. Cent. Eur. Agric., vol. 10, no. 3, pp. 283–288,
2009.
EP

[140] S. S. Arogba, “Mango (Mangifera indica) kernel: chromato- graphic analysis of the tannin, and
stability study of the associated polyphenol oxidase activity.,” J Food Compos Anal, vol. 13,
pp. 149–156, 2003.
CC

[141] K. Khanbabaee and T. V. Ree, “Tannins: Classification and Definition,” Nat. Prod. Rep., vol. 18,
pp. 641–649, 2001.
[142] D. W. Kelly, S. N. Paul, and J. E. Waller, “Method for flocculating and removing solids
suspended in water,” US 4,781,839, 1988.
A

[143] W. Vermerris and R. Nicholson, Phenolic Compound Biochemistry. Springer Science &
Business Media, 2006.
[144] M. Gaugler and W. J. Grigsby, “Thermal Degradation of Condensed Tannins from Radiata Pine
Bark.,” J. Wood Chem. Technol., vol. 29, no. 4, pp. 305–321, 2009.
[145] J. Lisperguer, Y. Saravia, and E. Vergara, “Structure and thermal behavior of tannins from
Acacia dealbata bark and their reactivity toward formaldehyde.,” J. Chil. Chem. Soc., vol. 61,
no. 4, pp. 3188–3190, 2016.
[146] G. O. Aspinall and A. Ca?as-Rodriguez, “810. Sisal pectic acid,” J. Chem. Soc., pp. 4020–4027,

41
 1958.
[147] D. G. Roux, D. Ferreira, H. K. L. Hundt, and E. Malan, “No Titl,” Appl. Polym. Symp, vol. 28, pp.
335–353, 1975.
[148] D. Mitchell et al., “Method of preparing a chemically modified tannin and its use,” EP 0 836
583 B1, 2003.
[149] S. E. Foster, “Treatment of manno galactan gums,” US 3346556 A, 1967.
[150] N. M. Levine, “Natural polymer sources,” in Poly-electrolytes for Water and Wastewater
Treatment, W. L. K. Schwoyer, Ed. Boca Raton, FL: CRC Press, 1981, pp. 47–60.
[151] M. Adinolfi et al., “Composition of the coagulant polysaccharide fraction from Strychnos
potatorum seeds,” Carbohydr. Res., vol. 263, no. 1, pp. 103–110, Oct. 1994.
[152] M. S. Otegui, “Endosperm cell walls: Formation, composition and functions,” Plant Cell

T
Monogr., vol. 8, pp. 159–174, 2007.
[153] C. S. Kumar and S. Bhattacharya, “Tamarind seed: properties, processing and utilization,” Crit.

IP
Rev. Food Sci. Nutr., vol. 1, pp. 1–20, 2008.
[154] S. Y. Choy, K. M. N. Prasad, T. Y. Wu, and R. N. Ramanan, “A review on common vegetables
and legumes as promising plant-based natural coagulants in water clarification,” pp. 367–390,

R
2015.
[155] D. Schwartz and R. L. Whistler, “History and future of starch,” in Starch: chemistry and

SC
technology, 3rd ed., J. N. BeMiller and R. L. Whistler, Eds. Elsevier Inc., 2009.
[156] M. Mandloi, S. Chaudhari, and G. K. Folkard, “Evaluation of natural coagulants for direct
filtration,” Environ. Technol., vol. 25, no. 4, pp. 481–489, 2004.

U
[157] A. Kibbe, “Guar gum,” in Handbook of Pharmaceutical Excipients, 6th ed., R. Rowe, P.
Sheskey, and M. Quinn, Eds. Pharmaceutical Press, 2009, pp. 298–300.
N
[158] A. A. Attama and O. J. Nwabunze, “Mucuna gum microspheres for oral delivery of
glibenclamide: In vitro evaluation.,” Acta Pharm., 2007.
A
[159] K. Gowthamrajan, G. T. Kulkarni, and A. Muthukumar, “Evaluation of Fenugreek mucilage as
gelling agent,” Int. J. Pharm. Expt., vol. 3, pp. 16–19, 2002.
M

[160] N. O. Ogaji I., “Film coating potential of okra gum using paracetamol tablets as a model drug,”
Asian J. Pharm., 2010.
[161] R. Singh, R. Malviya, and P. K. Sharma, “Extraction and Characterization of Tamarind Seed
D

Polysaccharide as a Pharmaceutical Excipient,” Pharmacogn. J., vol. 3, no. 20, pp. 17–19,
2011.
TE

[162] P. Somboonpanyakul, Q. Wang, W. Cui, S. Barbut, and P. Jantawat, “Malva nut gum. (Part I):
Extraction and physicochemical characterization,” Carbohydr. Polym., vol. 64, pp. 247–253,
2006.
[163] A. Noorlaila, A. Siti Aziah, R. Asmeda, and A. R. Norizzah, “Emulsifying properties of extracted
EP

Okra (Abelmoschus esculentus L.) mucilage of different maturity index and its application in
coconut milk emulsion,” Int. Food Res. J., 2015.
[164] N. K. Mathur, “Galactomannan polysaccharides,” in Industrial Galactomannan
CC

Polysaccharides, T. & F. Group, Ed. CRC Press, 2012.

[165] E. Anderson, “No Title,” Ind. Eng. Chem, vol. 41, p. 2887, 1949.
[166] I. C. M. Dea and A. Morrison, “Chemistry and interactions of seed galactomannans,” Adv.
Carbohydr. Chem. Biochem., vol. 31, pp. 241–312, 1975.
A

[167] C. Palanuvej, S. Hokputsa, T. Tunsaringkarn, and N. Ruangrungsi, “In vitro glucose entrapment
and alpha-glucosidase inhibition of mucilaginous substances from selected Thai medicinal
plants.,” Sci. Pharm., vol. 77, pp. 837–849, 2009.
[168] J. X. Jiang, L. W. Zhu, W. M. Zhang, and R. C. Sun, “Characterization of Galactomannan Gum
from Fenugreek (Trigonella foenum-graecum) Seeds and Its Rheological Properties,” Int. J.
Polym. Mater. Polym. Biomater., pp. 1145–1154, 2007.
[169] A. Wade and P. Weller, “G. gums,” in Handbook of Pharmaceutical Excipients, 2nd ed.,
London: The Pharmaceutical Press, 1994, pp. 215–216.

42
[170] A. Haddarah et al., “The structural characteristics and rheological properties of Lebanese
locust bean gum,” J. Food Eng., vol. 120, pp. 204–214, 2014.
[171] S. Kok, “A comparative study on the compositions of crude and refined locust bean gum: In
relation to rheological properties,” Carbohydr. Polym., vol. 70, pp. 68–76, 2007.
[172] E. J. Vernon-Carter, C. I. Beristain, and R. Pedroza-Islas, “Mesquite gum (Prosopis gum),” in
Novel macromolecules in food systems, G. Doxastakis and V. Kiosseoglou, Eds. Elsevier
Science B.V, 2000.
[173] E. J. Vernon-Carter and P. Sherman, “Rheological properties and applications of mesquite tree
(Prosopis juliflora) gum. L Rheological properties of aqueous mesquite gum solutions,” J.
Texture Stud., vol. 11, pp. 339–349, 1980.
[174] K. R. Bulusu and V. P. Sharma, “Pilot studies on the use of Nirmali seed as a coagulant aid,”

T
Environ. Heal., vol. 7, pp. 165–176, 1965.
[175] H. Mirhosseini and B. T. Amid, “A review study on chemical composition and molecular

IP
structure of newly plant gum exudates and seed gums,” Food Res. Int., vol. 46, no. 1, pp. 387–
398, 2012.
[176] P. V. Salimath and R. N. Tharanathan, “Carbohydrates of field bean (Dolichos lablab),” Cereal

R
Chem., vol. 59, no. 5, pp. 430–435, 1982.
[177] N. D. Zaharuddin, M. I. Noordin, and A. Kadivar, “The use of hibiscus esculentus (Okra) gum in

SC
sustaining the release of propranolol hydrochloride in a solid oral dosage form,” Biomed Res.
Int., 2014.
[178] D. McGarvie and H. Parolis, “The mucilage of Opuntia ficus-indica,” Carbohydr. Res., vol. 69,
no. 1, pp. 171–179, 1979.
U
[179] B. Matsuhiro, L. E. Lillo, C. Sáenz, C. C. Urzúa, and O. Zárate, “Chemical characterization of the
N
mucilage from fruits of Opuntia ficus indica,” Carbohydr. Polym., 2006.
[180] M. F. Saettone, S. Burgalassis, B. Giannaccini, and E. Boldrini, “Ophthalmic Solutions
A
Viscosified with Tamarind Seed Polysaccharides,” US6056950 A, 2000.
[181] P. Goyal, V. Kumar, and P. Sharma, “Carboxymethylation of Tamarind kernel powder,”
M

Carbohydr. Polym., vol. 69, pp. 251–255, 2007.

[182] H. Kaur, S. Yadav, M. Ahuja, and N. Dilbaghi, “Synthesis, characterization and evalution of
thiolated tamarind seed polysaccharide as a mucoadhesive polymer,” Carbohydr. Polym., vol.
D

90, pp. 1543–1549, 2012.

[183] J. Sánchez-Martín, J. Beltrán-Heredia, and J. A. Peres, “Improvement of the flocculation
TE

process in water treatment by using Moringa oleifera seeds extract,” Brazilian J. Chem. Eng.,
vol. 29, no. 3, p. 495, 2012.
[184] C. Ramamurthy, M. U. Maheswari, N. Selvaganabathy, M. S. Kumar, V. Sujatha, and C.
Thirunavakkarasu, “Evaluation of eco-friendly coagulant from Trigonella foenumgraecum
EP

seed,” Adv. Biol. Chem., vol. 2, pp. 58–63, 2012.

[185] S. A. Muyibi and L. M. Evison, “Moringa oleifera seeds for softening hard water,” Water Res.,
vol. 29, no. 4, pp. 1099–1105, 1994.
CC

[186] A. Ndabigengesere and K. Subba Narasiah, “Quality of water treated by coagulation using
Moringa oleifera seeds,” Water Res., vol. 32, no. 3, pp. 781–791, Mar. 1998.
[187] R. McBurney, C. Griffin, A. Paul, and D. Greenberg, “The nutritional composition of African
wild food plants: From compilation to utilization,” J. Food Compos. Anal., vol. 17, no. 3, pp.
A

277–289, 2004.
[188] B. Dawney, C. Cheng, R. Winkler, and J. M. Pearce, “Evaluating the geographic viability of the
solar water disinfection (SODIS) method by decreasing turbidity with NaCl: A case study of
South Sudan.,” Appl. Clay Sci., vol. 99, pp. 194–200, 2014.
[189] M. Lea, “Bioremediation of Turbid Surface Water Using Seed Extract from the Moringa
oleifera Lam. (Drumstick) Tree.,” in Current Protocols in Microbiology, N. J. Hoboken, Ed. John
Wiley & Sons, Inc.USA, 2014, p. (p. 1G.2.1-1G.2.8).
[190] K. A. Yongabi, D. M. Lewis, and P. L. Harris, “Integrated phyto-disinfectant-sand filter drum for

43
 household water treatment in Sub-Saharan Africa,” J. Environ. Sci. Eng., vol. 5, pp. 947–954,
2011.
[191] B. Garcia-Fayos, J. M. Arnal, V. Ruiz, and M. Sancho, “Use of Moringa oleifera in drinking
water treatment: study of storage conditions and performance of the coagulant extract,”
Desalin. Water Treat., pp. 1–7, 2016.
[192] A. T. A. Baptista et al., “Coagulation–flocculation process with ultrafiltered saline extract of
Moringa oleifera for the treatment of surface water,” Chem. Eng. J., vol. 276, pp. 166–173,
Sep. 2015.
[193] S. A. Muyibi and L. M. Evison, “Optimizing physical parameters affecting coagulation of turbid
water with Moringa oleifera seeds,” Wat. Res., vol. 29, no. 12, pp. 2689–2695, 1995.
[194] T. Okuda, A. U. Baes, W. Nishijima, and M. Okada, “Coagulation mechanism of salt solution-

T
extracted active component in Moringa oleifera seeds,” Water Res., vol. 35, no. 3, pp. 830–
834, 2001.

IP
[195] T. Radovich, “Farm and Forestry Production and marketing profile for Moringa (Moringa
oleifera),” in Specialty Crops for Pacific Island Agroforestry, C. R. Elevitch, Ed. Holualoa,
Hawai: Permanent 866 Agriculture Resources (PAR), 2009.

R
[196] A. Bonton, C. Bouchard, B. Barbeau, and S. Jedrzejak, “Comparative life cycle assessment of
water treatment plants,” Desalination, vol. 284, pp. 42–54, 2012.

SC
[197] K. C. Valverde et al., “Optimization of process conditions in water treatment through
coagulation diagrams, using Moringa oleifera Lam and aluminium sulphate,” Desalin. Water
Treat., vol. 56, no. 7, pp. 1787–1792, Nov. 2015.
[198]
U
S.-H. Kong, S.-K. Loh, R. T. Bachmann, S. A. Rahim, and J. Salimon, “Biochar from oil palm
biomass: A review of its potential and challenges,” Renew. Sustain. Energy Rev., vol. 39, pp.
N
729–739, Nov. 2014.
[199] N. M. Boopathi, “Moringa project,” Providing Economic Benefits to the Resource Poor Farmers
A
and Well-being for All: Collection, Conservation and Utilization of Moringa Germplasm.
[Online]. Available: https://sites.google.com/site/drnmboopathi/projects/moringa-project.
M

[200] F. Anwar and M. I. Bhanger, “Analytical characterization of Moringa oleifera seed oil grown in
temperate regions of Pakistan.,” J. Agric. Food Chem., vol. 51, no. 22, pp. 6558–6563, 2003.
[201] F. Anwar and U. Rashid, “Physico-chemical characteristics of Moringa oleifera seeds and seed
D

oil from a wild provenance of Pakistan,” Pakistan J. Bot., vol. 39, pp. 1443–1453, 2007.
[202] S. G. Zaku, S. Emmanuel, A. A. Tukur, and A. Kabir, “Moringa oleifera: an underutilized tree in
TE

nigeria with amazing versatility: a review,” Afr J Food Sci, vol. 9, pp. 456–461, 2015.
[203] J. K. Abaliwano, K. A. Ghebremichael, and G. L. Amy, Application of the Purified Moringa
Oleifera Coagulant for Surface Water Treatment, vol. 5. 2008.
[204] M. C. Wilbert, “Enhancement of membrane fouling resistance through surface modification.
EP

A study using the principle of membrane fouling and cleaning to develop ways to enhance
membrane fouling resistance,” in Water Treatment Technology Program Report No. 22,
Denver, CO, USA: US Department of the Interior, Bureau of Reclamation, 1997.
CC

[205] M. W. LeChevallier, T. M. Babcock, and R. G. Lee, “Examination and characterization of

distribution system biofilms.,” Appl. Environ. Microbiol., vol. 53, no. 12, pp. 2714–24, Dec.
1987.
[206] K. A. Ghebremichael, “Natural resources for appropriate water treatment,” in Sustainable
A

development of water resources, water supply, and environmental sanitation, 2006, pp. 434–
439.
[207] A. Arbenz and L. Avérous, “Chemical modification of tannins to elaborate aromatic bio-based
macromolecular architectures,” Green Chem., vol. 17, pp. 2626–2646, 2015.
[208] T. Nharingo and M. Moyo, “Application of Opuntia ficus-indica in bioremediation of
wastewaters: A critical review.,” J. Environ. Manage., vol. 16, no. 6, pp. 55–72, 2016.
[209] H. S. Stoker, “Carbohydrates,” in Organic and biological chemistry, 7th ed., Cengage Learning,
2015, pp. 292–293.

44
[210] G. Floridi, V. Langella, B. G. Li, R. Pelizzari, and U. Pfeiffer, “Hydroxypropyl guar,”
WO2012159972 A1, 2012.
[211] S. Subramanium, V. Nand, M. Maata, and K. Koshy, “Moringa oleifera and other local seeds in
water purification in developing countries,” Res. J. Chem. Environ., vol. 15, no. 2, pp. 135–
138, 2011.
[212] M. Asrafuzzaman, A. N. M. Fakhruddin, and M. Alamgir Hossain, “Reduction of turbidities of
water using locally available natural coagulants,” Int. Sch. Res. Netw. ISRN Microbiol., vol. 6,
2011.
[213] D. V. Kukic, M. B. Śćiban, J. M. Prodanović, A. N. Tepić, and M. A. Vaisć, “Extracts of fava bean
(Vicia faba L.) seeds as natural coagulants,” Ecol. Eng., vol. 84, pp. 229–232, 2015.
[214] S. Choubey, S. K. Rajput, and K. N. Bapat, “Comparison of efficiency of some natural

T
coagulants-Bioremediation,” Int. J. Emerg. Technol. Adv. Eng., vol. 2, no. 10, pp. 429–439,
2012.

IP
[215] N. J. Marobhe, G. Renman, and M. Jackson, “Investigating on the performance of local
materials for coagulation of turbid river water,” Inst. Eng. Tanzania, vol. 8, no. 3, pp. 50–62,
2007.

R
[216] M. Al-sameraiy, “A Novel Water Pretreatment Approach for Turbidity Removal Using Date
Seeds and Pollen Sheath,” vol. 2012, no. February, pp. 79–92, 2012.

SC
[217] A. Mukhtar, W. Ali, and G. Hussain, “A Preliminary Study of Opuntia stricta as a Coagulant for
Turbidity Removal in Surface Waters,” vol. 52, no. April, pp. 117–124, 2015.
[218] L. G. Torres, L. C. U. Sandra, and V. Mabel, “Use of Prosopis laevigata seed gum and Opuntia

U
ficus indica mucilage for the treatment of municipal wastewater by coagulation-flocculation,”
Nat. Resour., vol. 3, pp. 35–41, 2012.
N
[219] A. A. Al-Samawi and E. M. Shokrala, “An investigation into indigenous natural coagulant,”
Environ. Sci. Technol., vol. A-31, no. 8, pp. 1881–1897, 1996.
A
[220] K. Anastasakis, D. Kalderis, and E. Diamadopoulos, “Flocculation behavior of mallow and okra
mucilage in treating wastewater,” DES, vol. 249, no. 2, pp. 786–791, 2009.
M

[221] M. Sćiban, M. Klasnja, M. Antov, and B. Skrbić, “Removal of water turbidity by natural
coagulants obtained from chestnut and acorn.,” Bioresour. Technol., vol. 100, no. 24, pp.
6639–43, Dec. 2009.
D

[222] V. Patale and J. Pandya, “Mucilage extract of Coccinia indica fruit as coagulant-flocculent for
turbid water treatment,” Asian J. Plant Sci. Res., vol. 2, no. 4, pp. 442–445, 2012.
TE

[223] C. R. Schulz and D. A. Okun, “Treating surface waters for communities in developing
Countries,” JAWWA, vol. 75, pp. 212–223, 1983.
[224] M. Pritchard, T. Mkandwire, A. Edmondson, J. G. O’Neil, and G. Kululanga, “Potential of using
plant extracts for purification of shallow well water in Malawi,” Phys. Chem. Earth Parts
EP

A/B/C, vol. 34, pp. 799–805, 2009.

[225] S. A. Unnisa, P. Deepthi, and K. Makkanti, “Efficiency studies with Dolichos lablab and solar
disinfection for treating turbid waters,” J. Environ. Prot. Sci., vol. 4, pp. 8–12, 2010.
CC

[226] S. Vara, “Screening and evaluation of innate coagulants for water treatment: a sustainable
approach,” Int. J. Energy Environ. Eng., vol. 3, no. 29, 2012.
[227] K. A. Youngabi, “Studies on the use of medicinal plants and macro-fungi (lower plants) in
water and wastewater purification. Tre for life Journal,” in Proceeding of an E-Seminar, 2004,
A

pp. 1–14.
[228] J. Idris, A. Md Som, M. Musa, K. H. Ku Hamid, R. Husen, and M. N. Muhd Rodhi, “Dragon Fruit
Foliage Plant-Based Coagulant for Treatment of Concentrated Latex Effluent: Comparison of
Treatment with Ferric Sulfate,” J. Chem., vol. 2013, pp. 1–7, 2013.
[229] K. Qureshi, I. Bhatti, and M. S. Shaikh, “Development of biocoagulant from mango pit for the
purification of turbid water,” Sindh Univ. Res. J., vol. 43, no. 1, pp. 105–110, 2011.
[230] U. Gassenschmidt, K. D. Jang, and B. Tauscher, “Chemical properties of flocculant-active
proteins from Moringa oleifera Lam,” Biol. Chem. Hopper-Seyler, vol. 372, p. 659, 1991.

45
[231] M. B. Śćiban, M. Antov, and M. T. Klašnja, “Extraction and partial purification of coagulation
active components from common bean seed,” APTEFF, vol. 37, pp. 37–43, 2006.
[232] M. G. Antov, M. B. Śćiban, S. R. Adamović, and M. T. Klašnja, “Investigation of isolation
conditions and ion-exchange purification of protein coagulation components from common
bean seed,” APTEFF, vol. 38, pp. 1–19, 2007.
[233] G. N. Bidhendi, “Plantago Ovata Efficiency in Elimination of Water Turbidity,” J. Water Resour.
Prot., vol. 1, no. 2, pp. 90–98, 2009.
[234] B. Ramavandi, S. Hashemi, and R. Kafaei, “A novel method for extraction of a proteineous
coagulant from Plantago ovate seeds for water treatment purposes,” MethodsX, vol. 2, pp.
278–282, 2015.
[235] E. Bazrafshan, F. K. Mostafapour, M. Ahmadabadi, and M. A.H., “Turbidity removal from

T
aqueous environments by Pistacia atlantica (Baneh) seed extract as a natural organic
coagulant aid,” Desalin. Water Treat., vol. 56, no. 4, p. 977, 2015.

IP
[236] G. H. Ali, Hegazy, E. Badr, H. A. Fouad, and R. M. El-hefny, “Comparative Study on natural
products used for pollutants removal from water,” J. Appl. Sci. Res., vol. 5, no. 8, pp. 1020–
1029, 2008.

R
[237] Y. C. Ho, I. Norli, A. F. M. Alkarkhi, N. Morad, and I. Technology, “New Vegetal Biopolymeric
Flocculant : A Degradation and Flocculation Study,” Iran. J. Energy Environemnt, vol. 5, no. 1,

SC
pp. 2–3, 2014.
[238] S. A. Muyibi and C. A. Okuufu, “Coagulation of low turbidity surface water with Moringa
oleifera seeds,” Int. J. Environ. Stud., vol. 48, pp. 263–273, 1995.

U
[239] M. N. Alo, C. Anyim, and M. Elom, “Coagulation and antimicrobial activities of Moringa
oleifera seed storage at 3°C temperature in turbid water,” Adv. Appl. Sci. Res., vol. 3, no. 2,
N
pp. 887–894, 2012.
[240] M. L. Jodi, Y. Yahaya, and M. A. Sokoto, “The use of some plants in water purification,” vol. 1,
A
no. 4, pp. 71–75, 2012.
[241] M. S. M, C. S. G, and P. D. Raut, “Use of Moringa Oleifera ( Drumstick ) seed as Natural
M

Absorbent and an Antimicrobial agent for Ground water Treatment,” vol. 1, no. 3, pp. 31–40,
2012.
[242] P. N. Egbuikwem and A. Y. Sangodoyin, “Coagulation efficacy of Moringa oleifera seed extract
D

compared to alum for removal of turbidity and E. coli in three different water sources,” Eur.
Int. J. Sci. Technol., vol. 2, no. 7, pp. 13–20, 2013.
TE

[243] B. Ramavandi, “Treatment of water turbidity and bacteria by using a coagulant extracted
from Plantago ovata,” Water Resour. Ind., vol. 6, no. August, pp. 36–50, 2014.
[244] P. Sarker, M. Rahman, and N. J. Easha, “Potentiality of Tamarindus indica , Litchi chinensis
and Dolichos lablab Seeds as Coagulant for the Removal of Turbidity of Surface water,”
EP

Jahangirnagar Univ. Environ. Bull., vol. 3, no. January, pp. 25–33, 2014.
[245] N. Marobhe and G. Renman, “Purification of Charco dam water by coagulation using purified
proteins from Parkinsonia aculeata seed,” Int. J. Environ. Sci., vol. 3, no. 5, pp. 1749–1761,
CC

2013.
[246] S. C. N. S. Katayon, M. J. M. M. Noor, and A. G. L. Abdullah, “The effectiveness of M. oleifera
as primary coagulant in high rate settling pilot-scale water treatment plant,” Int. J. Eng.
Technol., vol. 3, no. 2, pp. 191–200, 2006.
A

[247] A. G. Liew, M. J. M. M. Noor, S. A. Muyibi, A. M. S. Fugara, T. A. Muhammed, and S. E. Iyuke,

“Surface water clarification using Moringa oleifera seeds.,” Int. J. Environ. Stud., vol. 63, no. 2,
pp. 211–219, 2006.
[248] J. Beltrán-Heredia and J. Sánchez-Martín, “Improvement of water treatment pilot plant with
Moringa oleifera extract as flocculant agent.,” Environ. Technol., vol. 30, no. 6, pp. 525–34,
May 2009.

46
All Figures
Captions

Figure 1: Trend of cumulative scientific publications on plant based

coagulants, and their use in water treatment from 1976 to 2017. Databases (Science Direct

(https://www.sciencedirect.com), National Centre for Biotechnology Information, NCBI

(https://www.ncbi.nlm.nih.gov/), ResearchGate (https://www.researchgate.net) , Mendeley

T
IP
(https://mendeley.com), Google Scholar (https://scholar.google.com) , Google

(https://google.com)), cross references and references within research articles were

R
SC
consulted to construct the graph.

Figure 2: Generalised structure of cationic starch.

Figure 3:
U
SSpro8 prediction of MO2.1 coagulant molecule.
N
Figure 4: Sequence alignment of cMoL and MO2.1 molecules. Underlined 1-Letter
A
amino acids show differences, whilst highlighted bold 1-Letter amino acids show similarities.
M

Figure 5: SSpro8 prediction of MusL coagulant molecule.

D

Figure 6: Generalised structure of condense tannin. Labels: ring ‘A’ represents

TE

‘resorcinol’; ring ‘B’ represents ‘pyrogallo’. (Source: Arbenz & Avérous [207]).

Figure 7: Generalised structure of poly-galacturonic acid. (Source: Nharingo & Moyo

EP

[208]).
CC

Figure 8: Generalised structure of tannin Mannich polymer. Labels: ring ‘A’ represents

‘resorcinol’; ring ‘B’ represents ‘pyrogallo’. Highlighted 6- and 8-positions of ring A show
A

attachment sites for primary amines. (Source: Arbenz & Avérous[207]).

Figure 9: Generalised structure of starch. (Source: Stoker[209]).

Figure 10: Generalised structure of Galactomannans (~1.1 Mannose:1 Galactose).

(Source: Floridi et al. [210]).

47
Figure 1:

180

160 Cumulative publications

140
Total No. of Publications

T
120

IP
100

80

R
60

SC
40

20

0
U
N
1970 1980 1990 2000 2010 2020
Year
A

Figure 2:
M
D
TE

CH2OH CH2O CHR`N+(CH3)3

EP

H
O O
H H
OH H OH H
CC

H H H

H OH H OH
A

Figure 3:

48
 T
Figure 4:

R IP
RRPAIQRCCQQLRNIQPRCRCPSLRQAVQLAHQQQGQVGPQQVRQMYRVASNIPA

SC
cMoL Sequence (3-57)

U
RQPDFQRCGQQLRNISPPQRCPSLRQAVQLTHQQQGQVGPQQVRQMYRVASNIPS
N
MO2.1 Sequence (5-59)
A
M

Figure 5:
D
TE
EP
CC
A

Figure 6:

OH

3̀ OH
2̀
4̀
1 1̀ B
8
OH 2
5̀ 49
7 OH
A 6̀
3
6
 5

OH

Figure 7:

T
IP
COOH COOH

R
H
O O

SC
H H
OH H OH H

H H H

H OH H U
OH
N
A
M

Figure 8:
OH
3̀
D

OH
2̀
TE

4̀
8 1 1̀ B
OH 2
5̀
EP

7 OH
A 6̀
3
CC

6 5 OH
4
CH2
OH
A

N R`

CH2

Figure 9:

CH2OH
50
 H H

H
OH H

O
H OH

CH2OH CH2 CH2OH

T
H

IP
O O O
H H H
OH H OH H OH H

R
H H H H H

SC
H OH H OH H OH

U
N
A
M

Figure 10:
D
TE

CH2OH CH2OH
H H Galactose
OH side group
OH
EP

H H
OH H OH H

H O H O
CC

H OH OH CH2 CH2OH
CH2
CH2OH
A

H O
O O O
H H
H H
OHOH OHOH
OHOH OHOH
H H H H H
H H
H H H H
H H H H

Mannose backbone
51
All Tables

Captions

Table 1: Overview on areas covered by published reviews

Table 1:

No. Histor Extract Coagulat Mecha Coagu Toxicity Upsc Applic Co Referen

T
of ical ion & ion nism lant / ale ation st ce

IP
pla use Purific effective prope Detrim
nts ation ness rties ental
D W effect

R
W W
      Bolto &

SC
Gregory
[30]
   Yongabi

9     U 
[75]
Yin [74])
N
1      Bichi
[73]
A
4   Theodor
o et al.
M

[72]
1        Kansal &
Kumari
D

[70]
     Choy et
TE

al. [71]
14   Choy et
al. [69]
11      Oladoja[
EP

68]
9      Oladoja[
67]
CC

   Nandini
&
Sheba[6
6]
A

       Kristiant
o[65]

52
Table 2: Overview on confirmation, identification and characterisation of active

compounds in extracts from known traditional plants. Symbol: sterisk (*) - partial

Table 2:

Traditional Verified Extractio Active Characteris Reference

coagulants through n compoun ed
lab process d
experimen optimise identifie

T
ts d d

IP
Arachis  Mbogo [120]; Subramanium et
hypogaea al. [211]

R
seeds
M.oleifera     Ghebremichael et al. [32];

SC
seeds Ghebremichael et al. [206];
Sánchez-Martín et al. [111];
Dezfooli et al. [29]
Parkinsonia
aculeata
   *
U Marobhe et al. [23]
N
seeds
Trigonella  Asrafuzzaman et al. [212]
A
foenum
M

graecum L.
seeds
Vigna    * Mbogo [120]; Marobhe et al.
D

unguiculata [23]; Subramanium et al. [211]

seeds
TE

Vicia faba  Kukić et al. [213]; Choubey et al.

seeds [214]
Voandzeia   Marobhe et al. [215]
EP

subterrane
an seeds
Phoenix  Al-Sameraiy [216]
CC

dactylifera
stones
Opuntia   Miller et al. [33]; Mukhtar et al.
A

sap [217]; Torres et al. [218]

53
Table 3: Brief list of plants explored as natural coagulants in water treatment

Table 3:

Plant Species Family Part Coagulant type References

Abelmoschus Malvaceae mucilage Neutral* Al- Samawi &Shokralla
esculentus/Hibiscus [219]; Gunaratna et al.
esculentus (okra) [119]; Anastasakis et al.
[220]; Zaharuddin et al.
[177]

T
Amorpha fruticosa Fabaceae seeds Unknown Sciban et al. [80]
(indigo bush)

IP
Aesculus Sapindaceae seeds Unknown Sciban et al. [221]
hyppocastanum

R
(Horse chestnut)
Cactus latifaria Cactaceae mucilage Neutral* Diaz et al. [22]

SC
Cassia alata Fabaceae leaves Unknown Aweng et al. [83]
(candlebrush)
Castanea sativa Fagaceae seeds Anionic* Sciban et al. [221];
(European chestnut) Živković et al [139]
Ceratonia siliqua
(carob; locust bean)
Fabaceae seeds
U
Neutral* Sciban et al. [80];
Haddarah et al. [170]
N
Cicer arietinum Fabaceae grains Unknown Asrafuzzaman et al.
(chick pea) [212]; Choubey et al.
A
[214]
Coccinia indica Cucurbitaceae Fruit Unknown Patale & Pandya [222]
M

(ivy gourd)
Cuminum cyminum Apiaceae seeds Unknown Ramamurthy et al. [184]
(cumin)
D

Cyamopsis Fabaceae gum Neutral Bratby [36]; Schulz &

psoraloides/ Okun [223]; Pritchard et
TE

Cyamopsis al. [224]

tetragonoloba (guar)
Dolichos lablab Fabaceae seeds Neutral* Unnisa et al. [225];
EP

(hyacinth bean) Asrafuzzaman et al.

[212]; Vara [226];
Choubey et al. [214]
Glycine max Fabaceae seeds Unknown Bhole [39]; Mbogo
CC

(Soybean) [120];
Hibisicus sabdariffa Malvaceae seeds Unknown Schulz & Okun [223];
(red sorella) Youngabi et al. [227];
A

Anastasakis et al. [220]

Hylocereus undatus Cactaceae fruit Unknown Idris et al. [228]
(dragon fruit) foliage
Inga edulis Fabaceae - leaf Unknown Obuzor et al. [82]
(ice-cream bean) Mimosoideae
Jatropha curcas Euphorbiaceae seeds Unknown Pritchard et al. [224];
(physic nut) Abidin et al. [25]
Lens culinaris Fabaceae seeds Unknown Schulz & Okun [223]
(lentils; pulse)

54
Mangifera Anacardiaceae Seeds Anionic* Qureshi et al. [229];
indica(mango) Arogba[140]
Manihot esculenta Euphorbiaceae Roots Unknown Vara [226]
Crantz syn. M.
utilissima
(tapioca sago)
M.oleifera(drumstick) Moringaceae Seeds Cationic Jahn & Dirar [107];
Gassenschmidt et al.
[230]; Ghebremichael et
al. [32]; Dezfooli et al.
[29]

T
Mucuna sloanei Fabaceae Seeds Unknown Menkiti et al. [81]
(mucuna bean seed)

IP
Opuntia ficus indica Cactaceae Mucilage Anionic & Miller et al. [33]; Torres
(nopal) neutral* et al. [218]; McGarvie &

R
Parolis [178]; Matsuhiro
et al. [179]

SC
Parkinsonia aculeate Fabaceae Seeds Cationic Marobhe et al. [23]
Phaseolus angularis Fabaceae seeds Gunaratna et al. [119]
(red beans)
Phaseolus mungo/ Fabaceae seeds Unknown Mbogo [120];
Vigna mungo
U Subramanium et al.
[211]
N
Phaseolus vulgaris Fabaceae Seeds Anionic Sciban et al. [80]; Sciban
(common beans; et al. [231]; Antov et al.
A
kidney bean) [232]
Phoenixdactylifera Arecaceae Seeds; Unknown Al-Sameraiy [216]
M

(date palm) pollen

grains
Plantago ovata Plantginaceae Seed Cationic Bidhendi et al. [233];
D

(psyllium Indian) Ramavandi et al. [234]

TE

Pleurotus tuber regium Pleurotaceae Sclerotium Unknown Youngabi et al. [227]

(king tuber
mushroom)
EP

Pistacia atlantica Anacardiaceae Seeds Unknown Bazrafshan et al. [235]

Pisum sativum Fabaceae Seeds Mbogo [120]
(Pea)
Prosopis juliflora Fabaceae Seeds Neutral* Diaz et al. [22]; Vernon-
CC

(mesquite) Carter et al. [172];

Vernon-Carter &
Sherman [173]
A

Prosopis laevigata Fabaceae Seeds Neutral* Torres et al. [218];

(smooth mesquite) gum Vernon-Carter et al.
[172]; Vernon-Carter &
Sherman [173]

Prunus Rosaceae Seeds Unknown Ali et al. [236]

armeniaca(apricot)
Red maize Poaceae Grains Unknown Gunaratna et al. [119]

55
Quercus robur Fagaceae Seeds Unknown Sciban et al. [221]
(common oak)
Quercus cerris Fagaceae Seeds Unknown Sciban et al. [221]
(turkey oak)
Quercus rubra Fagaceae Seeds Unknown Sciban et al. [221]
(Northern red oak)
Robinia pseudoacacia Leguminosae Seeds Unknown Sciban et al. [80]
(black locust)
Strychnos Loganiaceous Seeds Neutral Schulz & Okun[223];
potatorumLinn (Strychnaceae) Adinolfi et al. [86]
(nirmali)

T
Scaphium Sterculiaceae Seeds Neutral* Ho et al. [237];
scaphigerum(Malva Somboonpanyakul et al.

IP
nut or Taiwan sweet [162]
gum)

R
Tamerindous indica Fabaceae seeds Neutral* Schulz & Okun [223];
(tamarind) Goyal et al. [181]; Kaur

SC
et al. [182]
Trigonella Fabaceae Seeds Neutral* Schulz & Okun [223];
foenum(fenugreek) Ramamurthy et al.

U [184]; Mathur [164];

Jiang et al. [168]
N
Vigna unguiculata Fabaceae Seeds Cationic Marobhe et al. [23]
Zea may(corn) Poaceae grains Neutral Mandloi et al. [156];
A
Schwartz & Whistler
[155]
M
D
TE
EP
CC
A

56
Table 4: Commercial tannins use as coagulants in water treatments

Table 4:

Commercial Supplier Plant source Primary use Reference

tannin
Tanfloc TANAC (Brazil) Acacia mearnsii Coagulant aid Sánchez-Martı ́n
de Wild &Beltrán-
Heredia [114]
Acquapol SETA (Brazil) Acacia mearnsii Coagulant agent Sánchez-Martı ́n

T
de Wild &Beltrán-
Heredia [114]

IP
Silvafloc SILVATEAM Schinopsis Coagulant agent Sánchez-Martı ́n
(Italy) balansae alone or in &Beltrán-

R
combination Heredia [114]
with other

SC
inorganic
coagulants
KlarAid 6400 DEARBORN Mimosa extract Coagulant agent Kelly et al. [142]
Chemical (USA) alone or in

with U
combination
other
N
inorganic
coagulants
A
M
D
TE
EP
CC
A

57
 R I
SC
Table 5: Bench scale analysis of plant based coagulants in water treatments. Abbreviations: RW – river water, DW – dam water, TP – tap

U
water, GW –groundwater, PW – pond water, SW - surface water; TNTC - Too numerous to count

N
Table 5:

A
Coagulant Test Initial conditions Outcome Reference
Name Dosage sample Turbidit pH Microbial Other Turbidity Final Microbial Other findings

M
[mg/L] y [NTU] count [NTU] reduction [%]
M. 1mL/L TW(kao 105 7.6 - Alkalinity: 7 - pH: unaffected; Ndabigengesere
oleifera lin) 53mg/L; Alkalinity: & Narasiah [186]
seeds
D Conductivit
y: 154
unaffected;
Conductivity:
TE
µmho/cm; unaffected;
8mg SO4/L; Sulfate
0.4 mg (unaffected);
NO3/L 1 mg NO3/L
EP

M. 10 SW 35 - - 7 - - Muyibi & Okuofu

oleifera (+ 40 [238]
seeds mg/L
CC

alum)
M. 20’000 RW - - 3.0 x - - Total coliform= - Alo et al. [239]
oleifera 103MPN 2 x102
seeds total MPN/mL;
A

coliform Mesophilic
/mL; bacteria= 5 x
286 x 102 102 CFU/mL;
CFU Mesophilic
bacteria fungi= No
/mL; 70 x growth
102
CFUfungi
/mL

58
 R I
SC
M. 120 RW 632±3 7.45 - - 5±2.36 - pH 6.15±0.09 Jodi et al. [240]
oleifera ±0.2
pods

U
M. 150 GW 12.4 8± Total Alkalinity: 3.1±0.5 ; Total coliform= pH7.2± 0.5; Mangale et al.
oleifera 0.05 coliform: 4 130±0.1 1 x102 ± 0.57 Alkalinity:100±0 [241]

N
seeds x105 ± 0.57 mg/L; SPC/100mL & .28 mg/L;
SPC/100m Hardness: 5± 0.57 Hardness:

A
L & 190±0.57 MPN/100mL 100±0.57 mg
1600MPN/ mg CaCO3/L

M
100mL CaCO3/L
M. 16 RW 123.3 7.5 Total Hardness: > 85% reduction 96% and 94% - Sánchez-Martín
oleifera coliforms: 152 mg reduction of et al. [183]
seeds 8.0 x102 CaCO3/L total and
D MPN/100
mL; Faecal
faecal
coliforms, and
TE
coliforms: almost 100%
4.0 x102 reduction of
MPN/100 faecal
EP

mL; Faecal streptococcus

streptococ
ci: 1.4 x102
MPN/100
CC

mL
M. RW RW, RW RW RW RW RW RW RW Egbuikwem
oleifera 4.5 mL PW, 20.5 6.8 E.coli Alkalinity: 0.91; E.coli pH 6.9; &Sangodoyin[24
seeds PW GW PW PW (CFU/100 32 mg/L; PW (CFU/100mL): Alkalinity: 30 2]
A

6 mL 125 5.3 mL: TNTC Total 4.13; 33 mg/L; Total

GW GW GW PW Hardness: GW PW Hardness:
2.5 mL 10.7 7 E.coli 17.1 mg/L; 1.03; E.coli unaffected;
(CFU/100 PW (CFU/100mL): PW
mL): 57 Alkalinity: 8 pH: unaffected;
GW 20 mg/L; GW Alkalinity: 24
E.coli Total E.coli mg/L; Total
(CFU/100 Hardness: (CFU/100mL): Hardness:
mL): 21 17.1 mg/L; 2 unaffected;

59
 R I
SC
TW GW
Alkalinity: pH 7.1;
36 mg/L; Alkalinity: 40

U
Total mg/L;
Hardness: Total Hardness:

N
87.5 mg/L; unaffected
Strychnos 10mL/L PW 228±34 7.12 Total Alkalinity: 5±1 Total coliform: pH ~7; Ramamurthy et

A
potatoru coliform: 126±16 14±2 Alkalinity: 72±8 al. [184]
m seeds 111±25 mg/L MPN/100mL mg/L

M
MPN/100
mL
Natural 0.03-2 TW(cla 10- - - < 0.02 & 0.9 - - Özacar & Şengil
tannins (+ 1 y) 20FTU FTU [56]
mg/L
alum)
D
TE
1 (+ 2.5 100-300 7-8 - - <5FTU - pH= 6.7-7.1 Özacar & Şengil
mg/L FTU [137]
alum)
EP

Plantago 0.25 RW(kao 50-300 Streptococ > 95.6 Total Ramavandi [243]
ovate mg/L lin) cus %turbidity elimination of
seeds coliforms= reduction streptococcus
100≥x≤200 and fecal
CC

CFU/100 coliform
mL; Fecal
coliforms=
300≥x≤400
A

CFU/100m
L
Trigonella 10mL/L PW 228±34; 7.12 Total Alkalinity: 8±1; Total coliform: pH ~7; Ramamurthy et
foenum coliform= 126±16 55±8 Alkalinity: 74±9 al. [184]
graecum 111±25 CaCO3 MPN/100mL mg/L
seeds MPN/100 mg/L
mL
Cicer 100mg/ TW(kao 49-95 - Total - Turbidity Total coliform: - Choubey et al.

60
 R I
SC
arietinum L lin) coliform= (before 1.0X 102 [214]
seeds 1.05 X 103 filtration)=4.2 MPN/100mL
MPN/100 NTU & 9.3 NTU;

U
mL Turbidity (after
filtration)=3.6

N
NTU & 8 NTU
Dolichos 100 TW(kao 100 - Total - Turbidity Total coliform= - Choubey et al.

A
lablab lin) coliform= (before 1.1 X 102 [214]
seeds 1.05 X 103 filtration)=11.4 MPN/100mL

M
MPN/100 NTU; Turbidity
mL (after
filtration)=9.8N
TU
Opuntia
stricta
10 RW D
105 7.1 - - 15 - - Mukhtar et al.
[217]
TE
cladodes
Opuntia 5-55 Model 56-368 - - 5-7 - - Miller et al. [33]
spp. sap drinkin
EP

g water
Tamarind 100 RW 90.3 6.95 - TDS: 773 Turbidity - pH:7.37; Sarker et al.
us indica mg/L; COD: (before TDS: 60 mg/L; [244]
seeds 390mg/L filtration)=11.29 COD: 44mg/L
CC

NTU; Turbidity
(after
filtration)=
8.67NTU
A

Litchi 100 RW 90.3 6.95 - TDS: 773 Turbidity - pH7.37; Sarker et al.
chinensis mg/L; COD: (before TDS: 60 mg/L; [244]
seeds 390mg/L filtration)=4.95 COD: 243mg/L
NTU; Turbidity
(after
filtration)=
2.76NTU

61
 R I
SC
Parkinson 6 DW 810 7.4 - Alkalinity: 3 - pH: 7.2- 7.4; Marobhe &
ia 74 mg/L; alkalinity:50-55 Renman [245]
aculeate COD: mg/L; COD:

U
seeds 80mg/L 23mg/L
Corchorus 5 RW 632±3.2 7.45 - 6±2.45 - pH: 6.70±0.12 Jodi et al. [240]

N
tridens (+80 ±0.2
leaves mg/L 3

A
alum)
Hibiscus 5 RW 632±3.2 7.45 - - 5±1.71 - pH:6.85±0.05

M
Sabdariff (+80 ±0.2
a calyx mg/L 3
alum)

D
TE
EP
CC
A

62
 R I
SC
Table 6: Pilot scale analysis of plant based coagulants in water treatments. Abbreviations: CF= Clari-flocculator; RF=Roughing filters; SF=

U
Sand filters

N
Table 6:

A
M
Type of Dosage Type of Flow Properties Findings Reference
coagulant (mg/L) water rate
(m3/hr)

Strychnos 1.5 RW D - Turbidity= 1200- Turbidity= 22-29 NTU Bulusu et al. [174]
TE
potatorum (mg/L) (Yamuna, 1530 NTU
seeds plus 10 India)
mg/L
alum
EP

M. oleifera 75-250 SW 1 Turbidity= 400NTU Turbidity (after filtration)= 5NTU Sutherland et al. [89]
(Malawi)
M. oleifera varied RW 0.36 Turbidity= 21.6–479 At 10 mg/L alone and 10 mg/L plus Muyibi & Alfugara [90]
CC

(Serdang (Low= 21.5–36.3 24 mg/L alum

Selangor, NTU; Turbidity (low turbid samples)=3.8-
Malaysia) Medium=51.8–77.4 2.7 NTU, and 4.3- 1.5 NTU
NTU; High= 163- At 40 mg/L alone and 20 mg/L plus
A

394NTU); pH=6.9– 40 mg/L alum

7.5; Alkalinity= 85– Turbidity (medium turbid samples)=
119;zeta potential= 3.9-2.9 NTU and 3.6- 1.4 NTU
-21.4 to - 27.6 At 100 mg/L alone and 40 mg/L plus
60 mg/L alum
Turbidity (High turbid samples)=4.3-
1.9 NTU and 2.2-2.1 NTU
Zeta potential=–18.8 to –22.9 mV

63
 R I
SC
M. oleifera 35 Surface - Turbidity= 4.6 ± 1.6 Turbidity (after sand filtration)= 1.3 ± Ghebremichael [206]
water 0.9;
(Asmara, Turbidity (after pumice-sand dual

U
Eritrea) filtration)= 1.3 ± 0.5
M. oleifera 80-100 Surface - Turbidity= 17.8± 3.1 Turbidity (after sand filtration)= 6.4 ± Ghebremichael [206]

N
water 1.6; Turbidity (after pumice-sand
(Asmara, dual filtration)= 6.0 ± 2.3

A
Eritrea)
M. oleifera 100 Model 0.024 Turbidity= 200 ± 10 Average turbidity removal efficacy Katayon et al. [246]

M
turbid NTU; pH=6.4-6.5; (before filtration)= 46%; Average
water zeta potential= - turbidity removal efficacy (after
20mV filtration)= 99%
M. oleifera 40 & Stream
170 water D0.36 Turbidity= 74.6&
383; Alkalinity= 105
Turbidity= 3.9 NTU(or 2.9 NTU after 3 Liew et al. [247]
hr) & 3.2 NTU (or 2.8 NTU after 3 hr)
TE
mg/L (Malaysia) & 66 mg/L; pH =7.4
&7.3
M. oleifera 150 River 0.36 Turbidity=6.5 NTU; Effects CF RF SF Ali et al. [236]
EP

water Total bacterial

Turbidity 3.5 3.1 2.5
(Nile, counts= 6x102
Africa) CFU/mL; Total 9.3 x 1.3 x 1.6 x
Total coliforms= 1.1 bacterial 103 103 103
CC

x103 MPN/100mL; counts

Faecal coliforms= (CFU/mL)
2.7x 102 Total 360 240 160
MPN/100mL; Faecal coliforms
A

streptococci= 1.3 Faecal 90 90 34

x102 MPN/100mL coliforms
Faecal 60 26 35
streptococci
M. oleifera 150 River 0.54 Turbidity=6.5 NTU; Effects CF RF SF Ali et al. [236]
water Total bacterial
(Nile, counts= 5x103 Turbidity 3.5 3.1 2.5

64
 R I
SC
Africa) CFU/mL; Total 8.0 x 6.3 x 8.0 x
Total coliforms= 1.6 bacterial 102 102 102
x104 MPN/100mL; counts

U
Faecal coliforms= (CFU/mL)
3.4x 102 Total 160 240 300

N
MPN/100mL; Faecal coliforms
streptococci= 5 x102 Faecal 34 17 34

A
MPN/100mL coliforms
Faecal 21 50 53

M
streptococci
M. oleifera 5 mL/L River - Turbidity=123.3 Turbidity (before filtration)=an Beltrán-Heredia et al.
water NTU; Ph= 7.5; average 71.02% removed [248]
(Guadiana
, Spain) D Hardness=
CaCO3 mg/L; Total
152 Turbidity (after filtration)= 0 NTU
Faecal coliforms (before filtration)=
TE
coliforms= 8.0 x102 98.4% reduction; Faecal coliforms
MPN/100Ml; Faecal (after filtration)= 99% reduction;
coliforms= 4.0 x102 High reduction of both total coliforms
EP

MPN/100mL; Faecal and faecal streptococci observed

streptococci= 1.4
2
x10 MPN/100mL
Mangifera 30 River 0.36 Turbidity=6.5 NTU; Effects CF RF SF Ali et al. [236]
CC

indica water Total bacterial Turbidity 3.0 2.5 1.8

(Nile, counts= 2.3x103 Total 2.0 x 1.0 x 1.6 x
Africa) CFU/mL; bacterial 103 103 102
Total coliforms= 2.2
A

counts
x102 MPN/100mL; (CFU/Ml)
Faecal coliforms= Total 1.6 x 80 55
1.0x 102 coliforms 102
MPN/100mL; Faecal Faecal 78 45 14
streptococci= 2.3 coliforms
2
x10 MPN/100mL
Faecal 1.9 x 58 26
streptococci 102

65
 R I
SC
Mangifera 30 River 0.54 Turbidity=6.5 NTU; Effects CF RF SF Ali et al. [236]
indica water Total bacterial
Turbidity 3.0 2.5 1.8
(Nile, counts= 2.1x105

U
Africa) CFU/mL; Total 4.2 x 9.8 x 9.1 x
Total coliforms= 5.0 bacterial 105 104 103

N
x103 MPN/100mL; counts
Faecal coliforms= (CFU/Ml)

A
Not detected (ND); Total 8.6 x 3.4 x 1.2 x
Faecal streptococci= coliforms 103 103 102

M
1.3 x102 Faecal ND ND ND
MPN/100mL coliforms
Faecal 1.3 x 57 57

Prunus 30 River D
0.36 Turbidity=6.5 NTU;
streptococci
Effects
103
CF RF SF Ali et al. [236]
TE
armeniaca water Total bacterial Turbidity 3.0 2.7 2.0
(Nile, counts= 6.0x104
Total 2.8x 2.0x 1.6x
Africa) CFU/mL;
bacterial 104 104 104
Total coliforms= 7.0
EP

counts
x103 MPN/100mL;
(CFU/Ml)
Faecal coliforms=
8.0 x102 Total 5.3x 1.8x 17
coliforms 103 103
CC

MPN/100mL; Faecal
streptococci= 2.6 Faecal 5.3x 1.4x 9
3
x10 MPN/100mL coliforms 102 102
Faecal 1.6x 22 11
A

streptococci 102
Prunus 30 River 0.54 Turbidity=6.5 NTU; Effects CF RF SF Ali et al. [236]
armeniaca water Total bacterial Turbidity 3.0 2.7 2.0
(Nile, counts= 3.7x104 Total 1.1x 3.1x 3x
Africa) CFU/mL; bacterial 104 103 102
Total coliforms= 8.2 counts
x103 MPN/100mL; (CFU/Ml)
Faecal coliforms= Total 1.5x 1.9x 16

66
 R I
SC
1.0 x102 coliforms 103 102
MPN/100mL; Faecal Faecal 69 41 15
streptococci= coliforms

U
8.0x102 MPN/100mL Faecal 80 28 11
streptococci

N
Tanfloc 2 River - Turbidity=123.3 Turbidity (before filtration)=50-60% Sánchez-Martín et al.
(commercial water NTU; pH= 7.5; removed [59]

A
tannin) (Guadiana Hardness= 152 Turbidity (after filtration)= 0 NTU
, Spain) CaCO3 mg/L; Total Effects on hardness, total coliforms,

M
coliforms= 8.0 x102 faecal coliforms and faecal
MPN/100mL; Faecal streptococci not shown
coliforms= 4.0 x102
D MPN/100mL; Faecal
streptococci= 1.4
TE
2
x10 MPN/100mL
EP
CC
A

67
 R I
SC
Table 7: Semi-and full- process scale up analysis of plant based coagulants in

U
water treatments

N
Table 7:

A
Treatment plant Type of Dosage Type of water Flow rate Properties Outcomes References
coagulant (mg/mL) (m3/hr)

M
Thyolo Water M. oleifera 150 Surface water 16 Turbidity= ≥ Turbidity= 1NTU Sutherland et al.
Treatment (Malawi) 400NTU [89]
Works, Malawi
Integrated phyto- M. oleifera
disinfectant-sand D Not
specified
Pond
water(Nigeria)
- Turbidity=130.3
NTU; pH= 7.6; total
Before filtration
Turbidity=30.8
Yongabi et al.
NTU; [190]
TE
filter drum unit, aerobic mesophilic pH= 7; total aerobic
Nigeria bacteria counts= too mesophilic bacteria
numerous to count; counts= 3.5 x102
E.coli counts= 5.1 CFU/mL; E.coli counts=
EP

x103 CFU/mL; 5.7 x102 CFU/mL;

Coliform counts= Coliform counts= 4.3
7.3 x103 CFU/mL; x102 CFU/mL; Yeast
CC

Yeast counts= 2.4 counts= 7.5 x102

x103 CFU/mL; CFU/mL; Pseudomonas
Pseudomonas counts= 1.5 x102
counts= 1.1 x103 CFU/mL
A

CFU/mL After filtration

Turbidity=4.8 NTU; pH=
7.6; total aerobic
mesophilic bacteria
counts= 8 CFU/mL; E.coli
counts= 0.3 CFU/mL;
Coliform counts= 5.3
CFU/mL; Yeast counts=

68
 I
R
SC
9.3 CFU/mL;
Pseudomonas counts= 3
CFU/mL

U
N
A
M
D
TE
EP
CC
A

69