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Colorants through cane sugar production and refining (Part 1)

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 SPRI Conference 2008
Colourants through cane sugar production and refining
Luis San Miguel Bento
www.sucropedia.com

1 Introduction
Sugar colourants are water-soluble compounds present in sugar process products that
absorb light in the visible spectrum. They are one of the most important impurities in
sugar industry. Generally colourants are divided in two categories: colourants produced
by degradation of hexoses; and those presented in sugar cane, as cane pigments and
phenolics that can form high molecular weight colourants during processing. Due to
different physical and chemical conditions during processing, colourants can be formed,
removed or changed chemically. In this paper it will be described this behavior during
raw cane sugar processing and refining.

2. Origin and Properties of Sugar Colourants

Sucrose in sugar cane is not associated with coloured compounds. However, in cane exist
colourants precursors that during process can react developing yellowish to dark brown
compounds. Formation of colour in process varies with medium conditions as
temperature, pH, concentration and presence of precursors and impurities that can be
catalysts or colour inhibitors. In Table 1 is summarized the different colourants found in
sugar processing. Details on these colourants are described later.
Sugar colourants are normally characterized by their chemical composition: colour
(visible and UV); molecular weight (MW) (Table 2); charge, polarity and variation of
colour with pH (IV - Indicator Value = colour at pH 9/colour at pH4) (Table 3).
Colour in sugar products is usually given in IU – ICUMSA unities, absorbance at 420 nm
after filtration with 0.45 m membrane and correction to pH 7.0. This colour
measurement does not give complete information on chemical nature of colourants. UV
spectrum can complement this information.
 Smith, 1966, refers that colorants presenting high colour have molecular weights ranged
from 350 to over 50,000 Da (Dalton). Normally, molecular weight determination is done
through GPC - Gel Permeation Chromatography techniques.
According to the MW distribution, colourants are usually divided in the following
groups: Low MW (LMW), Medium MW (MMW), High MW (HMW) and Very High
MW (VHMW).
Low MW colourants, with MW lower than 2,500 Da include flavonoids, phenolic acids
and other monomeric phenols. This group includes also products of the reaction of
polyphenols with iron (Godshall, 2008).

Table 1 – Origin of sugar colourants

Reaction Reactant Sugar Colourant
Sugars Hexoses Degradation Products N compounds Melanoidins
OH - HADP
Heat Caramels
Phenols Enzymatic browning reactions Enz. BP *
Non – enzymatic browning reactions N compounds Non Enz. BP* (Melanins)
Oxidation Non Enz. BP*
Fe Iron-phenols complexes
No reaction Cell Wall Polysaccharides
Flavonoids
Phenolic acids
* Enz. BP / Non Enz. B – enzymatic / non enzymatic browning products (phenols browning products)

Medium MW colourants, are those with MW ranging from 2,500 to 12,000 Da, as HADP
and caramels.
High MW colourants, with MW ranging from 12,000 to 50,000 Da, include melanoidins
and products of browning reactions involving phenols. This group of colurants has dark
brown colour.
Very High MW colourants, with MW over 200,000 Da and passing through 0.45 m
membrane filter, comprise polysaccharides bonded to phenols or their oxidized products.
Lindeman, 2001, refers that VHMW colourants have a high carbonyl character indicating

2
a possible association with polysaccharides. Godshall, 2008, also refers that this
colourants are often associated with polysaccharides.
Other class of colourants, retained on 0.45 m membrane filter but passing through 0.80
m membrane filter are present in sugar solutions. The colour impart by these colourants
in sugar products was named by Godshall, 2005, as hidden colour.

Table 2 – Molecular weights of sugar colurants

Group VHMW HMW MMW LMW
MW > 200,000 Da 12,000 – 50,000 Da 2,500 – 12,000 Da < 2,500 Da

Enzymatic BP of CWP Melanoidins HADP Flavonoids

Phenols Brow. Prod HMF Polymers Phenolic Acids
Caramels Phenols-Iron

Variation of colour with pH is a simple way to make a rough idea about colourants nature
in sugar products. IV of main sugar colourants is presented at Table 3. Colourants
presenting higher IV are polyphenols, free flavonoids and their glucosydes (Riffer, 1988).
Normally HMW colourants have a lower IV, compared with LMW but some polymeric
colourants have high IV and some monomeric ones have low IV (Kennedy and Smith,
1976).

Table 3 - IV (Clarke et al. 1987) and pK (Riffer, 1988) of sugar colourants

Colourants IV pK

Phenols and flavonoids* 5.0 – 40 *

Fructose alkaline degradation products 1.5 – 3.2 5.1

Melanoidines 1.0 – 1.2 6.9

Caramels 1.0 – 1.5 4.7

* P. Smith, 1981

Colourants charge is an important characteristic of sugar colourants, especially when

ionic interaction is involved, as in resins. Colourants charge varies with medium pH and
colourant pK (Table 3). Most of colourants have five ionic charges per molecule (Riffer,
1988) and have an acidic character (Chou and Riztutto, 1992), being anionic in alkaline

3
conditions. Most of colurants are weak acids with an amphyphilic character (with anionic
and apolar zones).
Colourants shape varies with pH. At high pH or in the absence of salts, high molecular
colourants form a configuration of high dimensions, rod like, due to the mutual repulsion
of their ionic charges (Riffer, 1988). At low pH high molecular compounds concentrate
adopting a globular form.

3. Sugar Colourants – Hexoses Degradation Products

Sucrose and invert sugars, may react during sugar extraction and refining process. At
high and low pH, high tempature, in the presence of amino compounds and inorganic
compounds, hexoses can be degraded forming coloured compounds (Table 4). The main
colourants originated by reactions involving sugars are formed during extraction and
refining process. They are: melanoidins, HADP (Hexoses Alkaline Degradation
Products) and caramels. These compounds present a high molecular weight, between
(5,000 and 50,000) and a lower IV than natural sugar colourants (Table 2).

Table 4 – Effect of impurities on hexoses degradation reactions (Riffer,1988)

Low pH pH 6-8 High pH
Decrease Increase Increase Decrease Increase
Calcium x x
Sodium x x
Potassium x
Phosphate x
Citrate xxx
Iron xxx

Hydrolysis of sucrose results in glucose and fructose that can be degraded forming high
reactive compounds as HMF – hidroximethylfurfural, = 234 nm (Kort, 1978), that can
polymerize forming coloured compounds of high molecular weight. Iron catalyzes
degradation reaction of hexoses, particularly at 6 – 8 pH (Riffer, 1988). This author also
refers that phosphates present an important role in the inhibition of these reactions by
forming a complex iron-phosphate that predominates.

4
3.1. Melanoidins
Melanoidins are produced by the reaction between carbonyl compounds (aldehydes and
ketones) and amino compounds. In sugar solutions the main source of carbonyl
compounds are invert sugars besides the compounds resulting from alkaline degradation
of hexoses and polyphenol oxidation. This complex reaction, known as Maillard reaction,
produces high molecular weight compounds with an intense colour. It is believed that the
initial step of Maillard reaction is the formation of N-D glucosyl amino acids (Riffer,
1988). Formation of melanoidins are influenced by the nature and concentration of sugars
and amino compounds, temperature, pH and reaction time. The presence of impurities as
iron, copper and phosphates can catalyze the reaction (Ellis, 1959) Formation of
melanoidins take place mainly over 65º brix (Paton and McCowage, 1987). The velocity
of formation of these compounds is very slow in low brix solutions. As the brix increases,
the contribution of Maillard products to solution colour, increases (Paton and Mc
Cowage, 1987).
Empirical formula of melanoidins is C17-18 H26-27 O10 N with a molecular weight between
10,000 and 50,000 Da with an average of 28,000 ± 1,000 Da (Binkley, 1970).
Melanoidins present a low negative charge at neutral pH and a positive charge at low pH
(Davis, 2001). At pH 7.2, 66.7% of melanoidines are ionized (pK=6.9) (Riffer, 1988).
The spectra of melanoidins at pH 11 and pH 3 present two peaks at 286 nm and 326 nm
(Bento, 2002).
27000 77000
5 1 .6 m in
25000 5 0 .6 m in
67000
23000 4 4 .1

21000 57000
5 0 .0
19000 4 1 .0
microVolts

microVolts

47000
17000
3 9 .0
37000 5 1 .9
15000

13000 4 1 .8
27000
11000
17000
9000

7000 7000
0 10 20 30 40 50 60 0 10 20 30 40 50 60
R t m in . R t m in .

Figure 1: ELS diagram of Melanoidins Figure 2: ELS diagram of HADP

5
In Figure 1 it is presented the diagram obtained by GPC with ELS detector (Bento and
Sa, 1998). Melanoidins present peaks at 39.0; 4.0; 44.1; 50.0 and 51.6 min. retention
time, corresponding to 50,800, 36,000, 21,900, 7,700 and 6,100 Da, respectively.

3.2. HADP - Hexoses Alkaline Degradation Products

HADP are polymers of carboxylic acids, of brownish yellow colour, resulting from
monosaccharides degradation in alkaline conditions, passing through the formation of an
enediol anion (Bruijn, 1986). Alkaline degradation of glucose is initiated with a rapid
reaction forming intermediate not coloured compounds with a maximum absorbency at
270 nm and a molecular weight of 200 Da. These compounds polymerize forming
colourants with molecular weight between 1,000 and 5,000 Da (Parker, William, 1968).
The presence of iron catalyze HADP formation (Kort, 1978)
The spectrum of HADP presents a peak at 265 nm, at pH 11, and a peak at 267 nm, at pH
3 (Bento, 2002). In Figure 2 it is presented the diagram obtained by GPC with ELS
detector (Bento and Sá, 1998). HADP present peaks at 41.8, 50.6 and 51.9 min of
retention time, corresponding to a molecular weight of 31,400, 7,000 and 5,500 Da,
respectively. At a pH 7.2, 99.2% of the HADP are ionized (pK = 5.1) (Riffer, 1988).

3.3. Caramels
Caramels comprise a group of low anionic charge compounds resulting from sucrose
thermal decomposition, composed by HMF polymers. HMF does not present visible
colour, but when polymerized, it forms high intense colour compounds.
The reaction of caramels formation is much slower than those of melanoidins and HADP
(Mersad et al., 2000). By this fact these colourants have not great contribution in colour
of juices and syrups (Paton and McCowage, 1987). These complex reactions includes
formation of levoglucosan, from invert sugars, that pyrolize to branched chain species
(Rifffer, 1988).
For caramels formation, sugar temperature must be higher than their melting point
(185ºC) (Ahari and Genetolle, 1961). Although process temperatures are in average much
lower than this, it can happen that sugar be accidentally in contact with temperatures

6
higher enough to produce caramelization. Also, some impurity traces can catalyze
caramels formation (Riffer, 1988).

100000
51.9 min
90000

80000

70000

60000
microVolts

50000

40000

30000

20000

10000

0
0 10 20 30 40 50 60
Rt min.

Figure 3: ELS diagram of Caramels Figure 4: Dimerization of ferulic acid in CWP

(Clarke et al., 1988 )

Spectra of caramel at pH 11 and pH 3 present peaks at 229 nm and 285 nm (Bento,

2002). In Figure 3 it is presented the diagram obtained by GPC with ELS detector (Bento
and Sa, 1998). Caramels present one peak at 51.9 minutes of retention time,
corresponding to a molecular weight of 5,500 Da. Molecular weight of this colourant
increases with reaction time and some authors refers higher molecular weights, up to
50,000 Da and with radii approaching 50 Angstrongs (Riffer, 1988). At pH 7.2, 99.7% of
caramels are ionized (pK = 4.7) (Riffer, 1988).

4. Sugar Colourants – Phenols and Phenols Degradation Products

Sugar cane pigments are mostly chlorophylls, carotenes, xanthophylls and flavonoids.
The first three pigments are insoluble in water are easily separated during cane juices
clarification (Farber and Carpenter, 1972). Phenolic acids and other phenolic compounds
are also present in sugar cane and play an important role in sugar colour formation.
Phenolic compounds present a high IV and are involved in enzymatic and non enzimatic
browning reactions, and in reactions with iron, forming very dark compounds. Phenols

7
are 66% ionized at pH 9.7 but at pH 7.7 only 0,6% of ionization of flavonoids occurs
(Riffer, 1988).

4.1. Flavonoids
The presence of falvonoids in sugar cane depend on its variety, maturity, presence of
diseases and environmental and cultivation factors (Riffer, 1988). Their general chemical
structure can be represented by: C6 – C3 – C6. Five classes of flavonoids were found in
sugar cane: flavones, calchones, catechines and anthocianins (Smith and Paton, 1985).
Each one of these flavonoids has a characteristic oxidation degree of the C3 central chain.
Flavonoids, being soluble and with a low anionic charge, are not easily separated during
extraction and refining processes. Many flavonoids have been found in sugar cane being
their majority derived from flavones.
Flavones, present in all sugar cane varieties, are weak acids almost completely ionized at
pH 9 (Smith and Paton, 1985). Generally associated to glycosides they are stable at
milling conditions and following processes (Paton, 1992). These compounds present
stable anionic forms at high pH being also stable at moderate acid conditions (Smith and
Paton, 1985). They present a light colour in acid solutions but in alkaline solutions are
yellow or greenish yellow (Paton, 1992). The IV of theses compounds is high, attaining
values above 30 (Paton, 1992). More than twenty flavones were identified in sugar cane,
all of them derived from apigenine, tricine and luteoline (Smith and Paton, 1985). Tricine
based flavonoids have higher IV than luteolin flavonoids (Kennedy and Smith, 1976).
Another kind of cane flavonoids is the anthocianins. These compounds are responsible
for the red, blue and purple colours in plants. In sugar cane the red colour that can appear
in cane surface is due to these compounds. As anthocianins are coloured at acid
conditions and colourless at high pH they have an IV inferior to the unity. These
compounds are unstable at neutral or alkaline conditions and are decomposed by heat
(Paton, 1992). Heating the anthocianins at pH 7 they are partially decomposed, with loss
of a ring, originating colourless compounds (Smith and Paton, 1985).

8
4.2. Phenolic acids
Phenolic acids occurring in sugar cane are usually derived from cinamic acids (Farber e
Carpenter, 1972). They are the building blocks of flavonoids (Kennedy and Smith, 1976).
Phenolic acids are very difficult to remove during extraction and refining, reaching the
final product. As an example, chlorogenic acid, resulting from the esterification of caffeic
acid and quinic acid, was detected in white sugar (Farber and Carpenter, 1972). This can
be a factor for white sugar colour increase at high pH (Riffer, 1988). Ferulic acidis is
pale yellowish compound that can darken by oxidation.
Phenolic acids are normally linked to alcohols, to other phenols or to polysaccharides by
ester bonds. An example of this esterification is the one that occurs in CWP – Cell Wall
Polysaccharides (or ISP - Indigenous Sugar Cane Polysaccharides), composed by
polysaccharides, as arabinoxylan with glucoronic residues esterified with ferulic acids
(Figure 4). These high molecular complexes exits in cells membranes and have a plant
protection function, against external aggressions (insects, cuttings, etc.) (Clarke et al.,
1988). Being soluble and with a low anionic charge, these compounds are not removed in
most factory and refining processes, appearing in the white sugar (Clarke et al., 1988).
Due to their high molecular weight CWP cause great difficulties in sugar processing,
increasing sugar products viscosity, resins poisoning, difficulties in crystallization and
massecuite purging in centrifugals. Also, they decreases refining process efficiency and
increases energetic consumptions and affect sugar quality (Clarke et al., 1988).
The bonds of ferulic acid to polysaccharides are difficult to hydrolyze, and their
association persists during process (Clarke et al., 1988). Nevertheless, at high alkalinity
conditions, the de-esterification of these compounds may happen (Figure 4). Phenolic
colourants separated from polysaccharides, by this reaction, will have lower affinity to
sugar crystals. By this reason, this reaction provokes a colour increasing in sugar
solutions but a decreasing in the colour of the sugar crystallized from the resulting
solution (Clarke et al., 1988).

4.3 Oxidation of phenolic compounds

In the presence of enzymatic complexes, as PPO - polyphenol oxidase, in an oxidant
environment, polyphenols are oxidized, forming quinones. These compounds may react

9
with o-diphenols, amino acids, proteins (Mersad et al., 2000) or other quinones, forming
polymeric compounds. The formation of these enzymatic browning compounds in juices
depends on the cane variety, on type and concentration of phenolic compounds, and on
activity of enzymes involved in the oxidation reaction (Paton, 1992). Flavonoids derived
from apigenine and lutheoline are more prone to suffer enzymatic browning than those
derived from tricine (P. Smith and Paton, 1985). The concentration of flavonoids in
juice, especially those derived from lutheoline, with an o-phenol group, are important
markers for a possible browning colour formation (Paton, 1992). Cane varieties
containing these flavonoids and chlorogenic acid have potentialities for colour formation
during milling process.
Phenolic acids esterified with polysaccharides, when undergoing an enzimatic browning
reaction, may contribute for the formation of cross-links among the carbohydrates,
resulting in very high molecular weight complexes with a high colour. An example of
this kind of reaction is the enzimatic oxidation of ferulic acid in CWP (Clarke et al.,
1988) (Figure 4). These compounds have a high molecular weight, a low IV, are difficult
to remove and have tendency to enter the crystal during boiling (P. Smith, et al., 1981).
Most of the browning products from sugar beet, are insoluble in water (Wyse, 1971;
Paton, 1992). This can be the main reason why beet white sugar can be produced directly
from beet syrups and cane sugar not.
Phenols, as chlorogenic acid, may also be oxidized non enzimaticaly, through chemical
reactions. These reactions are slower than enzimatic ones, but their velocity may increase
at high pH and temperature (Paton, 1992). This kind of reaction can occur during sugar
solutions storage, even at low temperatures (Cilliers and Singleton, 1989; Paton, 1992).

4.4. Reaction of phenols with amino compounds

Penhols can react with amines forming compounds of intensive colour and high
molecular weight (> 2,500) named melanines. These compounds are difficult to remove
during refining.

10
4.5. Reactions of phenols with iron
Iron soluble in juices may form complexes with anthocianins, as with most of phenolic
compounds forming dark colourants of great stability (P. Smith and Paton, 1985; Riffer,
1988). This reaction may happen whenever sugar solutions are in contact with iron,
particularly during sugar extraction in mills. Complexes of iron with catechols are
believed to be responsible to the greenish hues in some brown sugars containing iron
(Riffer, 1985). These compounds present a low molecular weight (Godshall, 2008).

4.6. Phenols and polysaccharides

Association of phenolic compounds with polysaccharides is important for sugar industry
due to the chemical affinity of polysaccharides to sucrose.
These associations may occur:
- by formation of ester bonds between phenolic acids and polysaccharides;
- by bridges between polysaccharides, calcium and colourants;
- by sequestering poliphenols in polisaccharidse cavities (Haslam and Lilley, 1985).

5. Sugar Colourants through Cane Raw Sugar Mills

White sugar production from sugar cane implies two distinct stages: sugar extraction
from cane, in mills, producing raw cane sugar; and refining of raw sugar producing white
sugar. Recently, technological efforts were developed to produce white sugar directly
from sugar cane, without the refining step, as it is done in beet sugar industry. Until now
there is no industrial process to perform such objective.
Sucrose in sugar cane is extracted in mills or diffusers and juice obtained is clarified,
concentrated and crystallized to raw sugar. Colourants are formed during this process
according to medium conditions and concentration of colour precursors present in cane
juice. In Tables 5 and Figure 5 are summarized the brix, alkalinity and temperature
through the process and the main colourants produced in each stage.

5.1. Cane sugar extraction

Colourants in cane juice are mainly phenolic compounds free or bonded to
polysaccharides. Cane juice colour is yellowish, varying from pale to strong yellow, with

11
colours between 4,000 IU and 20,000 IU and an IV between 10 and 33. This colour is
influenced by phenolic compounds, as the high IV suggests.
Paton, 1992, deducted a good correlation (r2 = 0.94) between juice colour at pH 9, and
flavonoids and chlorogenic acid contents in juice (in ppm):
Colour @ pH 9 = 6,267 + 83.7 x [Flavonoids] + 18.6 x [Chlorogenic acid]
As can be observed, flavonoids have a great influence in juice colour at pH 9.
When milling is used for sugar extraction, juice colour is affected by mill operation.
Higher extractions results in higher juice colour (P. Smith et al., 1981). During milling,
besides sugar, polysaccharides (originates from cane or formed after harvesting) and cane
pigments including flavonoids and other phenolic compounds are also extracted. In the
mill tandem colour and flavonoids increases from the first mill to the last one (Paton,
1992).
P. Smith et al, 1981, reported a colour variation from first expressed juice to mixed juice,
from 23,000 to 38,300 in one Australian sugar mill. This represents an increase of 66% of
the first expressed juice colour. An example of colourants through the tandem is
presented at Table 6.

Table 5 - Main colourants formed in each step of the process

Production Extraction Clarification Evaporation Sugar House
Products Cane Juice Juice Syrup Syrup
Massecuites
Molasses
Sugars
brix Low Low Low/High High
Alkalinity Low High Medium Medium
Temperature Medium High High Medium
Colurants CWP Enz. Brow. P. HADP Melanoidins Melanoidins
Flavonoids Phenols-Iron Non Enz. BP Non Enz. BP
Phenolic acids Caramels Caramels

Along the mill, flavonoids concentration increases from 50 ppm - 150 ppm, on solids, to
values until 2,000 ppm in the last mill (P. Smith and Paton, 1985). Separation of the

12
juices from the last mills to produce ethanol, as is done in Brazil, will decrease mix juice
colour and the raw sugar production will be more economical.
During milling there is a decrease of chlorogenic acids concentration, neutral phenolics
and flavonoids (Table 7) indicating the formation of enzymatic browning products (4.3).
According to Goodacre and Coombs, 1978, this reaction contributes to more than half of
colour in cane juices. Paton, 1992, refers a value of 53% of the first extraction juice
colour, at pH 7, been originated by this oxidation reaction.

Figure 5 – Colourants formation through mill process.

In the last decades the utilization of cane diffusion to replace the potent roller mills has
occurred in different part of the world. Juice obtained with this extraction system presents
a higher colour than the one obtained by milling. Probably enzymatic browning reactions
during diffusion produce more colourants, due to the higher residence time and more
contact with air during the extraction process.

Table 6 – Colours (pH 7) along the mill tandem

1st expressed juice 14,300
nd
2 “ 32,300
3rd “ 45,900
th
4 “ 62,600

13
5.2. Juice clarification
In clarification, calcium hydroxide is added to mixed juice with heating at a temperature
around 100 oC. The lime will neutralize organic acids and inorganic ions as phosphates
and sulfates. At high pH and high temperature, enzymes will be denatured. PPO and
other enzymes cease their activity and formation of enzymatic browning colourants will
cease.
In this stage the behavior of phenolic compounds varies according to their chemical
structure. Chlorogenic acid concentration increases in this operation. Paton, 1992, refers
an increase from 60 to 160 ppm, on juice solids. The same is observed to the cinnamic
acids like caffeic, p-cumaric and ferulic acids (from 50 to 110 ppm). These concentration
increases can be due to the hydrolysis of esters bonds of these acids with polysaccharides
as in case of CWP. This hydrolysis happens due to high alkalinity conditions occurred
during clarification (Figure 4). Also, during clarification anthocianins are decomposed in
residual phenolics (P. Smith, Gregory, 1971). Due to the release of phenolic acids, by
these two ways, juice IV increases, from 4.6 into 6.9 (Paton, 1992). Due to these
reactions, there is a redistribution of colourants molecular weights in juice, to low
molecular weight compounds (P. Smith et al, 1981). Other consequence of these
reactions is the fact that Curtin and Paton, 1980, found a higher concentration of phenolic
acids in raw sugar than in cane juice.

Tabela 7 – Concentration of phenolic compounds in juices (Paton, 1992)

Cane juice Mixed juice Variation

Chlorogenic acids * 800 ppm 300 ppm - 62%
Neutral phenolics 310 ppm 110 ppm - 64%
Flavonoids 1070 ppm 380 ppm -64%
* Chlorogenic acid and isomers

Some flavones are soluble and stable at high pH and, therefore, will be not destroyed in
clarification (P. Smith and Paton, 1985).
Colourants are formed in clarification: some phenols react with amino compounds
(Paton, 1992) and, due to the high alkalinity, HADP colourants will be formed. Neutral
phenolics suffer a 35% decrease, from 200 ppm into 130 ppm, on juice (Paton, 1992)

14
possibly consumed on the reaction with amino compounds. Paton and Mc Cowage, 1987,
report that HADP formation increases proportionally with invert sugars concentration and
with pH increase.
However, during clarification, there is some decolourization. Paton, 1992, observed a
colour decrease of 13% (from 16,200 to 14,100 IU). P. Smith et al., 1981, refers a colour
removal during clarification as being between 20 and 25%. Deducting the colour
formation referred, the actual colour removal during clarification must be higher.
According to Tu et al., 1977, in clarification there is a reduction of more than 50% of
high molecular compounds. P. Smith et al., 1981, report a decrease of polymeric
colourants in clarification. Favonoids are not well removed with lime (Farber and
Carpenter, 1972).

5.3. Evaporation
Clarified juice is normally concentrated in five bodies evaporators. Concentration
increases from 15º brix until approximately 65º brix. In this satge colourants are formed.
P. Smith et al, 1981, reported a colour increase during evaporation of 6% between
clarified juice and liquor (syrup).
Non enzymatic oxidation reaction of phenols can occur. The velocity of this reaction is
lower as the one of enzymatic browning reactions (Paton, 1992). Also, due to brix
increases Maillard reactions can occur with formation of melanoidins. This reaction
depends mainly on amino compounds concentration and on other factors as pH, brix and
invert sugars concentration (Paton and McCowage, 1987).

5.4. Crystallization
Sugar crystallization is processed in different steps: concentration, seeding (or magma
seed in beginning), crystal growing and thickening. In the end of the boiling massecuite is
discharged to crystallizers and then centrifuged to separate sugar from mother syrup.
Colourants in massecuite comprise the ones in feed syrup, in magma seed and colourants
formed during boiling and during massecuite storage in the crystallizers. The presence of
melanoidins during boiling increases viscosity of syrups and massecuites interfering with
crystallization (Kort, 1978).

15
Colourants formed during boiling are especially caramels (Kennedy and P. Smith, 1976)
and melanoidins (Honig, 1959). P. Smith, et al. 1981, reports an increase of massecuite
colours of 9%, 14% and 19% in A, B and C massecuites, respectively, during boiling and
storage in crystallizers. Munday et al., 1968, report colour formation higher than 30% in
storage of C massecuites. This colour increases is mainly due to Maillard reaction
compounds
During boiling, in the crystal formation and growth phases, colourants remain mainly in
mother syrup but a small quantity will be integrated in crystal. Some colourants presents
high tendency to be incorporated in crystal. Many studies have been made to quantify this
tendency. The term colour transfer, the ration between sugar (or crystal) colour to feed
liqour (or massecuite) colour is used to quantify this tendency (Chen and Sloane, 1980;
Lionnet, 1988; Donnavan and William, 1992). Paton, 1992, defines the partition
coefficient as the ratio of massecuite (or liquor) colour and sugar colour.
Lionnet, 1995, related crystal colour, after affination the sugar in the laboratory at
standard conditions, with feed liquor colour. The slope of the resulting line determines
the colour transfer. Lionnet, 1995 found colour transfer of 0.0190 for liquors from
carbonatation - sulfitation; 0.0231 for liquors from carbonatation - ion exchange and
0.0241 for liquors from phosphatation - ion exchange. This author also conclude that:
- higher crystallization rates increases colour transfer;
- colour transfer increases with crystal size;
- extracts of green cane tops, mixed with feed, increases colour transfer;
- an increase of HADP in feed do not increase colour transfer.
Other authors, Mantovani, 1986; Donavan and William, 1992; Mann, 1987 and Vaccari
et al., 1991, arrive also to some of these conclusions.
The correlation between crystal colour and crystal width is given by the following
equation (W is the mean crystal width in m):

Crystal colour = - 124 + 0.0194 x feed colour + 0.388 x W

Using GPC, Bento et al., 1997, separated colourants and other compounds, as
polysaccharides, in raw sugar, using a Superose 12 column with a spectrophotometer

16
DAD and ELS- evaporative light scattering detector. In Figures 6 and 8 are presented the
chromatograms of raw sugar and affined sugar, and in Figures 7 and 9, the respective
ELS diagrams. Comparing with ELS diagrams of synthetic colourants (Figures 1,2 and 3)
we can assume that peaks B1, B2 and B3 are compounds from melanoidins group; C1 is
a typical peak from HADP and C2 is a peak common to melanoidins and caramels, being
probably a HMF polymer. Peak B3 is not present in any synthetic colourant diagram and
could be from phenols browning products.

60 80000
B4

50
C1
60000
40
Abs.x1000

MicroVolts
B3
30 40000 C2

20

20000
A
B2
10 B1

0 0
0 10 20 30 40 50 60 70 80 0 10 20 30 40 50 60 70 80
Rt (min) Rt (m in)

Figure 6–Chromatogram,330 nm) of Raw Sugar Figure 7 - ELS diagram of Raw Sugar

ELS diagram do not detect low molecular weight compounds, as these were eluted jointly
with sucrose and other sugars.

70

80000
60

50
60000

40
Abs.x1000

MicroVolts

30 40000

20
20000
10

0 0
0 10 20 30 40 50 60 70 80 0 10 20 30 40 50 60 70 80
Rt (m in) Rt (m in)

Figure 8–Chromatogram (330nm) of Affined Sugar Figure 9 - ELS diagram of Affined Sugar

The evaluation of HMW and MMW colourants in raw sugar and affined sugar (crystal)
was made measuring their peaks heights. The percentage of the occurrence of these
colourants (Peaks B2, B3, B4, C1 and C2) in raw sugar and affined sugar are resent in
Table 9. In this Table is presented the ratio of these percentages, colourant % in affined

17
sugar / colourant % in raw sugar (% on total colourants of this group). To this ratio we
name TR – Transfer Ratio. In Figure 10 it is presented TR of these colourants as a
function of respective molecular weight. This curve gives the relative tendency of these
group colourants to be integrated in the crystal.
Transfer rate (TR) of a given colourant (X) can be defined more exactly as the ratio
between mass of colourant X / mass of all colourants, in the crystal (sugar affined in
standard conditions) and sugar (raw sugar, white sugar or other crystallized sugar). That
is: (Xcrystal / T crystal) / (X sugar / T sugar). This calculation must involve the largest spectrum
of sugar colourants we can found in sugar products: those in the previous example and
the phenols and their derivatives. As it will be difficult to evaluate the total mass of
colourants in a sugar or crystal (T sugar or crystal), a standard colourant must be found (S).
This standard must be a single compound present in all sugar products that in a test with
the universe of known colourants will give a TR of 1.0. Then TR can be calculated as
(Xcrystal x S sugar) / (X sugar x S crystal).

It is generally accepted that high molecular weight colourants have more affinity to the
crystal (Tu et al., 1977; Roberts and Godshall, 1978; P. Smith et al, 1981; Clarke et al.,
1984; Mantovani, 1986; Lionnet, 1991; Saska, 1991. Donnovan and William, 1992,
found that treating feed liquors with membranes, removing HMW colourants, colour
transfer is reduced. Roberts and Godshall, 1978, report that washed raw sugar has a
higher percentage of HMW colourants than the original raw sugars and Kennedy and P.
Smith, 1976, reported that there is an increase of polymeric colourants in the affined
sugar, compared with raw sugar. Godshall et al., 1987, refers that 2/3 of colourants
remaining in affined sugar have a MW higher than 20,000 Da. By the results presented in
Table 8 and Figure 10, we observe that the integration of colourants, ranging from 5,000
to 36,000, into crystal will increase with molecular weight. Tu et al., 1977, refers that low
MW (< 5,000) tends to be excluded from sugar crystal. These authors also showed that,
although high molecular weight colourants are present in a lower quantity in juices, they
have a high influence in crystal colour. The fact that high molecular colourants with low
IV have a higher tendency to sugar explains the decrease of IV value between clarified

18
juice and sugar obtained from it. From a juice with a colour of 14,100 IU and an IV of
6.9, it was produced a raw sugar with 2,300 IU and an IV of 4.6 (Paton, 1992).
Kort, 1978, also refers that melanoidins have some tendency to be incorporated on crystal
surface, but a higher tendency to be included inside the sugar crystal. In Table 8 it is
observed that peaks B2 has the higher TR meaning a higher occurrence in crystal than in
sugar. This peak corresponds to melanoidins of 36,000 Da. Other peak of melanoidins,
B4 - 21,900 Da, has a lower TR.

Table 8 – Percentage of peak heights Ratio

4.0

Peak Raw Sugar Affined Sugar AS/RS

3.0
B2 2.1 % 7.8 % 3.7
B3 18.9 % 29.0 % 1.5 2.0

B4 34.2 % 35,8 % 1.0

1.0
C1 28.5 % 20.9 % 0.7
C2 16.2 % 6.6 % 0.4
0.0
0 10000 20000 30000 40000
MW

Figure 10 – Ratio of ELS peaks (Table 8)

N. H. Smith, 1976, showed that enzymatic browning colourants enter the crystal during
boiling. Peak B3 – 28,000 Da, can be due to phenols browning reactions. This peak shoes
a tendency to be integrate in the crystal (Table 8).
Kort, 1978, and Riffer, 1988, refers that HADP and caramels, during boiling, will be
integrated in the external layer covering the crystal. Bento et al., 1997, showed that after
washing a white sugar crystal, the peak of HADP and caramels decreases (Figures 19 and
20). Peak C1 in Figure 7, with 7,000 Da, is characteristic of HADP. This peak shoes an
TR lower than 1.0 meaning their tendency to be incorporated in the sugar external layer
(Table 8). Also, peak C2 – 5,500 Da, characteristic of caramels and HMF polymers,
shoes the lowest TR of all colourants in Table 8.

Low molecular weight colourants include plant pigments, namely monomeric phenolics
and phenolic acids. Phenolic colourants can be ordered by rising integration capacity as
follows: phenolic acids, as chlorogenic acid, then flavonoids, and finally, neutral

19
phenolics. Colourants originated from enzymatic browning reactions present high affinity
to the crystal (Paton, 1992). After affination this raw sugar in a high speed centrifugal
with separation of 2% m/m of syrup, Paton, 1992, observed, through HPLC analysis, that
over half the chlorogenic acid, cinamic acid and flavonoids of raw sugar were present in
the syrup, but only 30% of neutral phenols were separated from the crystal. Paton, 1992,
also observed, using synthetic colourants, that HADP and melanoidins present a higher
partition coefficient to the crystal (feed colours/crystal colour) than cane juice colourants.
This means that these colourants present a lower integration capacity to the crystal as
compared with cane juice colourants.

Integration mechanism of sugar colourants into crystals is not clear. It is possible that
colourants with high affinity to sucrose molecule have a higher integration capacity. For
example phenols linked to polysaccharides, flavones with sugar residues or their
oxidation products. Also it is possible that colourants with a high carbonyl nature can for
bridge with polysaccharides through calcium ions. It is known that calcium in presence of
sucrose enhance resins regeneration possibly by formation of a complex sucrose-calcium
colourant (Bento, 1996). Another parameter, after chemical affinity, that influence
colourants integration into crystals is the colourant molecular weight (Figure 10) that
affects diffusion of colourants in syrup during cryatallization.

Table 9 – Integration of colourants in raw sugar

In the crystal CWP

Enzymatic browning products

Melanoidins

Neutral phenols

In the external layer Melanoidins 21,9 kDa

HADP
Caramels
Flavonoids
Phenolic acids

20
Integration of colourants in crystal happens after seeding, in the crystals growing step.
Other kind of colourants will be integrated in sugar crystal during the thickening phase.
In this phase, supersaturation and crystallization rate increases, and colourants will be
integrated in the external surface layer. In Table 9 is presented the distribution of main
sugar colourants in the internal crystal and on external sugar layer.

5.5. Raw sugar

Raw sugar is the result of first crystallization in the sugar factory (A boiling). Sucrose
content in a raw sugar is, normally, between 98,5% and 99,5%. This sugar presents a
brownish yellow colour and a crystal size about 1mm MA (mean aperture). N. H. Smith,
1966, estimated that colourants represent 16 – 20% of non sugars in a raw sugar with
98.5 Pol. Raw sugar quality has evolved from 96º Pol in the sixties of last century, till the
VHP (very high pol) raw with a Poll higher than 99º. In Australia, a raw sugar type was
produced, the VLC sugar, presenting a high Pol and a low colour (Burns and Field,
1998). The objective of VLC production is to transfer to the mills, at lower energy costs,
the most part of purification process. In the refineries, this sugar will need only to be
melted, filtered and crystallized.
An important characteristic of raw sugar colourants is their distribution between crystal
and the external layer. A good raw sugar is the one that, after affination, presents a low
colourants content in the affined sugar. The knowledge of the type of colourants in the
crystal is important in order to evaluate their behavior during refining.
Observing the Figures 6, 8 and 11, representing GPC chromatograms at 330 nm of raw
sugar, affined sugar and affination syrup, (Bento, 1997) we can calculate the relative
absorbencies of the three MW groups of colourants (Table 10). The ratio of colour
(absorbancies at 330 mn) percentages of affined sugar to raw sugar is presented in the last
column. As specific absorbancies of colourants are different, these values are not
comparable with those obtained with the quantities of colourants. However, it is
important to note the increase of crystal colour due to low molecular colourants (mainly
phenols).

21
Table 10 – Distribution of colourants absorbencies (330 nm) Table 11 – ELS peaks distribution Raw
Sugar and Affination Syrup
Raw Sugar Aff. Sugar Aff. Syrup ASg/RS
Peak Aff. Sug. Aff. Syr. Sg/Sy
HMW 51% 39% 55% 0.76
MMW 27% 23% 29% 0.85 B2 7.8 % 2.5 % 3.1

LMW 22% 38% 16% 1.73 B3 13.8 % 13.8 % 2.1

B4 32.2 % 32.2 % 1.1
C1 32.5 % 32.5 % 0.6
C2 19.0 % 19.0 % 0.3

Paton, 1992 calculated that 70% of a raw sugar colour, recently crystallized, without
storage, is originated by high molecular weight colourants (measured at 405 nm). In
Table 10 we observe that for raw sugar, colourants (at 330 nm) higher than 2,000 Da
represents 78% of total colourants.
P. Smith and Paton, 1985, refer a presence of phenolic compounds, flavonoids type, in
Austalians raw sugars between 10 ppm and 60 ppm, contributing to 30% of sugar colour,
at pH 7. Lindeman, 2001, refers that Australian raw sugars present a percentage of 50 –
60% in weight of HMW colourants in the crystal and these colourants have the double of
the colour of LMW colourants.
Kennedy and P. Smith, 1976, reported that 75% of colour in raw sugar is localized in the
surface layer. Godshall et al., 1987, report that affination removes 70% of total raw sugar
colour. Making a colour balance on colourants in raw sugar, affined sugar and affination
syrup (Table 11) it is observed that an average of 73% of colourants (at 330 nm) are in
the syrup layer of this raw sugar. When magma seed is used for A boiling, crystal colour
of raw sugar is influenced by C crystal colour (Munday et al., 1968).
During storage raw sugar colour can increase, depending on storage time, temperature,
relative humidity and colourants and impurities in the syrup layer. Impurities in raw sugar
are important for colour formation in storage. Petri and Carpenter, 1979, reported that
colour of a low quality raw sugar duplicated in one year storage at 30ºC, but a good
quality sugar increased only 50% in the same conditions. VH Pol sugars form less colour
during storage has they have a lower syrup layer, compared with normal raw sugars
(Kort, 1978).

22
Colour formation is limited to the syrup layer. In a raw sugar, syrup in the surface layer
has a high brix and, therefore, is a factor for colour formation, by Maillard reactions, with
loss of amino nitrogen (Paton, 1992). Phenols, as chlorogenic acid, may also be oxidized
non enzimatically, through chemical reactions. Paton, 1992, reported that a raw sugar
with 3,740 IU colour, stored during 20 days at 30ºC, in an atmosphere of 52% of RH,
suffered a colour increase of 42%. She observed too that cinnamic and chlorogenic acids
have decreased 50%, from their initial values of 12 ppm and 23 ppm, respectively. With
atmospheres of 63% of RH, more drastic changes occurred. After 10 days of storage, raw
sugar colour increased 57% and phenolic acids concentration decreased to 1 ppm.
Phenolic acids concentration decrease can be explained by their oxidation (Paton, 1992).
This is in accordance with the decrease of IV during raw sugar storage. In the examples
referred above, IV decreased from 3.7 to 2.7 at 52% RH, and to 2.1 at 63% RH. Neutral
phenols and flavonoids, in the same example, do not suffer a great change. As neutral
phenols are inside sugar crystals, they are not exposed to the external oxidants conditions,
as the ones in the external syrup layer. It was proposed that 0,05% of invert sugars
present in raw sugar can suffer caramelisation catalyzed by inorganic compounds,
especially carbonates, during storage (Farber and Carpenter, 1970).
To minimize colour formation, raw sugar must be cooled to below 32 oC before storage
(Kort, 1978).

6. Sugar Colourants through Cane Raw Sugar Refineries

In the refining process colourants in raw sugar can be removed, transformed or increased
by colourants formation during process. In Table 12 are presented the compounds MW
distribution (higher than 2,500 Da) through refining process (Bento et al., 1997). N. H.
Smith, 1966 referred that colourants from refinery products are in the range of 350 to
50,000 molecular weight and that IV increases through affination, clarification and
crystallization. Roberts and Godshall, 1981, refers that 46 to 93% of colour in refinery
products has a molecular weight higher than 12,000 Da.

23
6.1 Affination
The distribution of colourants in raw sugar (crystal and external layer) was discussed
before (5.5). A raw sugar containing a low quantity of colourants with low integration
capacity, will be a good sugar for affination, if other characteristics were normal, as grain
size, resulting in a low colour affined sugar. In Affination the majority of colourants in
raw sugar are separated to the affination syrup. A decolourization of circa 62%, in
relation to raw sugar colour, is achieved in this operation (Godshall et al., 1987).
By the previous discussion (5.5) it is observed that high molecular compounds from the
raw sugar will be preferentially retained in the affined sugar.
In the ELS diagram of affination syrup (Figure 12) it is observed VHMW colourants
decreased, as compared with raw sugar diagram. The distribution of HMW and MMW
colourants are similar to raw sugar. As it is observed in the last column in Table 11, the
ratio between percentages of colourants in affined sugar and affination syrup are similar
to their TR values (relation between affined sugar an raw sugar) (Table 8).

Table 12 – Molecular weight compounds distribution (>2,5 kDa) through refining

> 250 kDa 12kD - 50 kDa 2,5kD - 12 kDa

Raw sugar 2.8% 55.7% 40.6%

Affined sugar 9.2% 70.3% 20.5%

Affination syrup 0.8% 50.7% 48.5%

Affination liquor 17.7% 37.8% 44.5%

Carbonatation liquor 17.5% 8.3% 73.4%

Fine liquor 19.1% - 80.9%

White sugar 55,1% 3,6% 41,3%

Washed white sugar 70.4% 2.2% 27.4%

Molasses 0.8% 39.6% 59.7%

Low MW colour increased in affined sugar, in relation to raw sugar, from 22 % to 38 %

on total colour at 330 nm (Table 10). This shows that low molecular weight compounds,
with high IV, will be retained in affined sugar after affination. This fact explains the

24
increase of IV through Affination, as from 3.0 IV to 3.6 IV (Kennedy and P. Smith,
1976).

70
60000
60

50
40000

MicroVolts
40
Abs.x1000

30
20000
20

10
0
0 0 10 20 30 40 50 60 70 80
0 10 20 30 40 50 60 70 80 Rt (min)
Rt (min)

Figure11–Chromatogram of Affination Syrup Figure 12 – ELS diagram of Affination Syrup

Affination is very efficient to remove colourants in raw sugar syrup layer. After
centrifugation, affined sugar must be melted and in this operation colourants, caramels,
can be formed. Also, other colourants are mixed with those of affined sugar: HADP from
sweet waters returned from clarification, and melanoidins and phenols browning products
coming with recovered sugar from recovery section. By these facts, colourants ELS
diagram of affination liquor is different than affined sugar diagram (Figure 9 and 13).

140000
90000
C1
C1
80000 B
120000
C2
70000
100000

60000
microVolts

80000
m ic r o V o lts

50000 A

40000 60000 C2

30000 A
40000
B
20000

20000
10000

0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
Rt min. R t m in .

Figure 13 – ELS diagram of Affination Liquor Figure 14 – ELS diagram of Carbonated Liquor

25
6.2 Clarification
Clarification in sugar refining is usually made through three processes: carbonatation,
phosphatation and filtration with filter aid. Although decolourization is not the main
objective of clarification, there is some colourants removal is this stage. Also, some
colorants are formed.
Carbonatation
Due to high alkalinity in the mixture of calcium hydroxide with affination liquor, invert
sugars decompose and HADP colourants are formed (Kenneth, P. Smith, 1976; Bento,
1999). Bento, 1999, reported that invert sugars were reduced through Carbonatation,
from 0.10% in affination liquor to 0.04% in carbonated liquor. This corresponded to an
increase on peak C1 in ELS diagrams (51% on MMW colourants) (Table 13 -
quantification done with an internal standard of polyethylene glycol ; Figure 15 ). It was
also observed that this formation is manifested in the second reactor. This fact can be due
to the reaction time for hexoses decomposition subsequent polymerization.
As it was as proved, HADP are difficult to remove from ion exchange resins and they
remain in white sugar (Bento and Sá, 1997). Due to these facts it is very important to
avoid the formation of these compounds during Carbonatation. Bento, 1999, proposed a
pre-carbonatation of one part of the lime (40%) to avoid an excess of alkalinity in the
lime mixing tank. With this process, decolourization during carbonatation increased from
51% to 53% due to a reduction of HADP colourants (Figure 15).

180.000,00

140,000.00
160.000,00

140.000,00 120,000.00

120.000,00 100,000.00

100.000,00
MicroVolts
MicroVolts

80,000.00

80.000,00
60,000.00

60.000,00
40,000.00

40.000,00
20,000.00

20.000,00
-
48.00 49.00 50.00 51.00 52.00 53.00 54.00 55.00
-
10,00 20,00 30,00 40,00 50,00 60,00
Rt (min.) Rt (min.)

Figure 15: ELS diagram of Affination Figure 16 – Reduction of peak C1 due to lime
Liquor (bold) and Carbonated Liquor pre-carbonatation (bold) (lower peak – Affination
Liquor; upper peak - Carbonatation Liquor)

26
Although this colour formation, there is a substantial colour reduction in Carbonatation,
over 50%. Colourants that have acidic nature can form weak bonds with calcium and will
be incorporated in the calcium carbonate precipitate during growing (Davis, 2001).
Kennedy and P. Smith, 1976, refer that polar polymeric colourants, with amino nitrogen
and anionic at high pH, have high affinity to the calcium carbonate precipitate.
Compounds of molecular weight between 12 kDa and 50 kDa, including melanoidins, are
substantially removed (Figure 15) (Bento and Sá, 1998).

Table 13: Compounds of high molecular weight in liquors

> 250 kDa 12kD – 50 kDa 2,5 – 12 kDa

Affination liquor 229 ppm 490 ppm 576 ppm

Carbonated liquor 211 ppm 99 ppm 870 ppm

Fine liquor 212 ppm --- 896 ppm

Compounds of very high molecular weight, higher than 250 kDa, suffer a reduction of
8% (Table 13, Figure 15). It is also observed that peak B3 in affination liquor disappear
after carbonatation. We suspect that this peak is due colourants bounded to
polysaccharides or to phenolic browning products (enzymatic or non enzymatic). In fact,
due to the high alkalinity observed during carbonatation, a hydrolysis of phenolic acids
esterified to polysaccharides can occur. Due to this hydrolysis, the coloured part, that is
the phenols or their oxidation products, will be separated from the carbohydrates, and
consequently, their integration to sugar crystals will decrease. This fact can explain that
white sugars obtained from carbonated liquors presents a lower colour than those from
phosphatation liquors of equivalent quality (Clarke et al., 1988).
Due to browning products hydrolysis, single phenols are released to carbonatation liquor.
Kennedy and P. Smith, 1976, refer that carbonatation removes few cane pigments of low
molecular weight. Farber, et al, 1968, refer that flavonoids are not well removed by
Carbonatation, although those derived from tricine are preferentially removed (P. Smith
and Paton, 1985).

27
Phosphatation
Phosphatation does not remove as much colourants as carbonatation. However, adding
cationic surfactants during phosphatation, colour removal will improve.
Phenolic aminated compounds are more removed in Carbonatation than in Phosphatation
(Clarke et al., 1988). When cationic surfactants are used in a clarification process, as
phosphatation, they fix to anionic colourants that precipitates and will be scavenged by
the calcium phosphate precipitate.

6.3. Decolourization
After Clarification liquor presents a colour of 600 ± 100 IU. This colour is not low
enough to produce a white sugar of 45 IU in a three boiling system. Excess of colourants
must be removed before crystallization to obtain a fine liquor colour of 250 UI at least.
Therefore, a decolourization higher than 60% must be done to the clarified liquor. In
order to perform this decolourization, different processes can be applied:
- using adsorbents carbons (bonechar, activated carbon granular or in powder);
- using ion-exchange (anionic resins; anionic polymers).
- by oxidation (ozone; hidrogen peroxide);
- by reduction (sulphitation; sodium hydrosulphite).
Colourants that persist in final liquor will depend on decolorization technique applied.
For example, granular carbons remove preferentially flavonoids derived from apigenin
and tricin; bone char those derived from tricin; and resins those derived from apigenin (P.
Smith and Paton, 1985). Refined sugar produced with liquors decolourized with resins
presents a higher proportion of polymeric colour than the ones produced with char
(Kennedy and P. Smith, 1976).

Adsorbent Carbons
Carbon was the first decolourizer in sugar industry. Due to their large surface area, from
500 to 2,000 m2/g (Davis, 2001), due to their porosity, they have a great decolourization
capacity. Adsorbent carbons can be powder activated carbons, bone char and granular
activated carbons (GAC). Colourants are adsorbed through van de Waals and London
forces that strongly fix colourants to the carbon surface.

28
Bone char remove all type of colourants, preferentially monomeric colourants, with high
IV, (N. H. Smith, 1966) as trycin-based flavonoids (Kennedy and Smith, 1976). These
authors refer also that char removes less efficiently low IV colourants, cane polymeric
colourants and factory colourants. The presence of high quantities of phosphates in liquor
may inhibit adsorption of iron complexes by char (Riffer, 1988).
Recently, char was almost completely replaced by granular activated carbons. These
carbons present a specific surface area ten times higher than char, and consequently, their
decolourizing capacity is much higher (Field and Benecke, 2000). GAC are very efficient
in removing flavonoids and other phenolic colourants. Flavonoids are more efficiently
removed by granular carbons than by bone char. Bardwell et al., 1985, refers that GAC
removes practically all flavonoids contained in liquors. Paton and P. Smith, 1983, refer
that GAC have high affinity to flavonoids but not to amino nitrogen derivatives
compounds. Lindeman, 2001, refers that virgin GAC removes 35% and 40% of
melanoidins (alkaline, and acid, respectively), 76% of HADP, 78% of melanoidins
(asparigine), 100% of caramels and 52% of HMW colourants. Davis, 2001, refers that
HMW colourants can be fixed through different molecule sites which turns the adsorption
stronger.
Granular carbons (Char and GAC) are regenerated thermically in kilns. This is a very
expensive operation, due to the high price of the kiln. Some refineries make GAC
regeneration in kilns separated from the refinery. Recently some trials on chemical
regeneration have been made (Kato and Fukui, 1993; Moodley et al., 2000; Bento, 2006).
Bento proposed a chemical regeneration using ethanol, sodium hydroxide and hydrogen
peroxide, with promising results. This process if applied in a cane sugar mill producing
ethanol is very advantageous as ethanol from the distillery can be used, and the
regeneration effluents can be sent back to fermentation. By this process the
dcolourization process have no effluents disposal.

Ion Exchange Resins

The great majority of cane colourants presents an amphyphilic (containing ionic and
polar zones) character. The fixation of these compounds to resins can be done through
different mechanisms:

29
- by ionic bond between ammonium quaternarium pole, fixed to resin, and anionic
colourant;
- by hydrophobic inter-action between colourant apolar part and resin matrix;
- by weak bonds as by van der Waals forces.
These mechanisms can act separately or in conjunction within the same colourant
molecule (Williams and Bhardwaj, 1988). Colourants containing aromatic groups will be
preferentially adsorbed by styrenic resins as these resins have aromatic groups in their
structure. However, removal of these compounds during salt regeneration will be more
difficult. Another phenomenon that can occur is the switch between fixation mechanisms,
due to the composition variation of the solution that surrounds the resin. This fact occurs
mainly with pH variation and during resin regeneration when NaCl concentration change
from 100g/l to almost 0g/l (Bento, 1992). When styrenic resins are contacted with
carbonated liquor containing increasing quantities of NaI, decolourization decreases as
iodide ion compete with colourants for the exchanges sites (Bento, 1992) (Figure 17). If
liquor pH is decreased, from pH 9 to pH 3, decolourization increases. This is due to the
exchange of fixation mechanism from ion exchange to hydrophobic interaction between
colourant and resin matrix. Most anionic colourant at a pH 3 will be in a neutral form (pK
in Table 3) Fixation of colourants by hydrophobic inter-action, depends on resin matrix
nature. Styrenic resins present a hydrophobic matrix while acrylic resins have a matrix
more polar. Styrenic resins are more efficient decolourizers than acrylic resins, however
are more difficult to regenerate.
Colourants formed during extraction and refining processes, are differently fixed and
removed from styrenic resins. Guimarães et al., 1999, using synthetic colourants (HADP,
melanoidins and caramels), observed that these colourants present different behaviors
during decolourization and resin regeneration (Table 14). By these results it is observed
that styrenic resins remove efficiently HADP and melanoidins. These resins do not so
efficiently remove caramels. This experiment was done with a new resin. Using a resin
not virgin the results will be different, especially in the case of HADP and caramels, as
explained below in regeneration. Lindeman, 2001 refers that resins remove 82% of
melanoidins, 69% of HADP, 57% of melanoidins (asparigin), 38% caramels and 34%
HMW colourants. Rader and Anderson, 1971, refer that resins are highly efficient to

30
remove HADP but less efficient for melanoidins and caramels. Kennedy and Smith,
1976, report that factory colourants are poorly removed. However, lowering the pH
decolorization increases, at expenses of other anionic colourants that will be then less
removed.
% Dec.
120000
C1
100
100000

80 C2
80000

m i c r o V o l ts
60
60000

40
40000

20 20000

0 0
0 10 20 30 40 50 60
0 0.5 1 1.5
R t m in .
NaI mol/L

Figure 17: Decolorization with styrenic Figure 18: ELS diagrama of Fine Liquor
in presence of NaI at pH 3 and pH 9 (bold) (after resins)

Bento and Sa, 1998, using GPC chromatography with an ELS observed that compounds
of molecular weight between 12,000 Da and 50,000 Da, (melanoidins), were totally
removed by resins (Table 13; Figure 18). It also observed that MMW colourants (HADP,
caramels) are practically maintained.
Table 14 - Colourants behavior during decolourization with resins

Decolourization Regenerated

HADP 98,0% 50,9%

Melanoidins 97,5% 90,8%

Caramels 62,8% 52,9%

During resin regeneration, with NaCl at 50g/l and 100g/l, melanoidins are practically
totally removed (Table 14). However HADP and caramels are not well removed. This
fact can be explained by the switch effect between the two fixation mechanisms (Bento,
1992). At the end of regeneration, NaCl concentration decreases and anionic colourants
fixed to the matrix will be released and will replace chloride ions bonded to resin. At the
beginning of the working cycle, stronger anions dislocate caramels and HMF polymers

31
from the resin (Bento and Sa, 1988) This can be avoided doing an acid wash at the end of
regeneration (Bento, 1992).
Kennedy and P. Smith, 1976, refer that resins have a high capacity to remove trycin
based colourants and low capacity to luteolin derivatives. Riffer, 1986, observed that
styrenic resins are more efficient that GAC to remove iron complex colourants. It is
thought that resin fouling is due to phenolic compounds mainly in combination with iron
(Kennedy and P. Smith, 1976)

Cationic surfactants
Cationic surfactants are also used to remove colourants from sugar solutions. Colourants
react with surfactant forming an insoluble precipitate.

Oxidation reactions
In some factories, oxidants, as ozone and hydrogen peroxide, are applied as decolourizers
(Mendoza and Espejo, 2002). Oxidants form active free radicals that react with functional
groups of colourants. At low pH oxidants, as ozone, form oxygen active radicals and, at
high pH, form hydroxyl radicals. Sucrose molecule can be destroyed by hydroxyl radicals
and the utilization of oxidants at high pH may be dangerous. However, Gomez et al,
1980, refer the destruction of sucrose only occurs at pH higher than 8.5.
Oxidants, as H2O2, decolourize by cleavage of conjugated non saturated bonds forming
carboxilic acids, increasing the anionic character of some colourants. This fact is
beneficial for carbonatation and resins application. Bento, 2004, treated, with hydrogen
peroxide, sugar liquors before strong base styrenic resins and noted that: resin efficiency
increased; resin will be less contaminated; resin cycles extended 3 or 5 times; less
effluent will be produced; fewer chemicals and utilities will be consumed; less sweet
water will be produced; operating costs with resin regeneration and effluent treatment
will be reduced.
Davis et al, 1998, demonstrated that ozone react with high IV colourants as flavonoids
and other phenols. Oxidants not only decompose coloured compounds but also colour
precursors as amines and phenols (Davis, 2001). Phenols are oxidized to quinones and
acyclic compounds (Riffer, 1988)

32
Reduction reactions
The application of reducing compounds in sugar industry is another way to reduce sugar
colour. In sulfitation process, sulfur dioxide is bubbled through the juice, liquor or syrup,
promoting the reducing of colourants that become colourless. In this reaction double
conjugated bonds, that absorb visible radiation, become saturated.
Sulfitation is more efficient in suppressing colour formation than as a decolourizer
(Davis, 2001). Caramels and melanoidins formation are inhibited by combination of the
sulfur dioxide with the reduction sugars blocking the carbonyl function (Zerban, 1947).
Hydrosulfites decolorize by reducing ketones, kinones and iron complexes. Kinones
reduction impedes their polymerization to HMW dark colourants (Riffer, 1988).
Bisulfites react with non saturated carbonyls and some flavonoids, dislocating the light
adsorption from the visible to the UV zone (Riffer, 1988). The action of sulfites provokes
antocianins decolourization being the colour regenerated through acidification (P. Smith
and Paton, 1985).

6.4. Crystallization and White sugar

The knowledge of colourants composition in white sugar is important in order to
maximize their removal during processing. In section 5.4 it was discussed the colourants
integration in sugar during crystallization. Here it is presented the case of white sugar
crystallization. Sugar colourants in granulated sugar represent about 30 ppm (N. H.
Smith, 1976). The ELS diagram of white sugar from a carbonatation-resins refinery is
presented in Figure 19 (Bento et al., 1997). Two groups of compounds are observed:
compounds with molecular weights higher than 250,000 Da, VHMW, corresponding to
polysaccharides and CWP, of low colour; and compounds with molecular weight from
2,500 Da to 12,000 Da, MMW, corresponding to HADP and caramels. (ELS diagram
does not shows LMW colourants). After white sugar washing with methanol, it was
observed that compounds of VHMW are maintained and that MMW compounds were
reduced, mainly the peak corresponding to caramels (Figure 20). This confirms the lower
transfer ratio of HADP and caramels to crystal and their presence mainly in the external
layer (Table 8).

33
 20000 22000

19000
20000
18000
18000
17000

16000 16000
MicroVolts

MicroVolts
15000
14000
14000

13000 12000

12000
10000
11000
8000
10000

9000 6000
20 25 30 35 40 45 50 55 60 20 25 30 35 40 45 50 55 60
Rt (min) Rt (min)

Figure 19: ELS diagram of white cane sugar Figure 20: ELS diagram ELS of cane white
sugar (after washing)

Lionnet, 1995, found that degrading carbonated liquor with lime, increasing liquor
colour from 612 to 812, did not increase crystal colour in the sugar produced with this
liquor. The colourants formed at high alkalinity, HADP, remain mainly in the crystal
surface and are removed after sugar affination. Bardwell et al., 1985, observed that, after
washing an amount of 10% of cane sugar crystals, colour reduction did not decrease after
further washing. Also they observed that the remaining colour, in crystal, corresponds to
60% of original sugar colour (for sugars of 1st, 2nd and 3rd boiling). Kennedy and
Smith, 1976, report a value higher than 50% for colour remaining in sugar crystal.
Farber and Carpenter, 1972, detected phenols, as ferulic acid, in white sugar. Kennedy
and P. Smith, 1976, report that in white sugar boiling there is an IV increasing from the
massecuite and to the sugar crystal. These authors indicate the possibility of integration
of flavonoids of high IV in white sugar and conclude that, in the conditions of CSR
refineries, colourants originated from plant pigments are the major contribution to sugar
colour. Also, N.H. Smith, 1976, reports that colourants with high IV have tendency to be
incorporated in sugar crystals during crystallization.
In trials made by Clarke et al., 1988, sugar produced with liquor where ferulic acid was
added, developed a yellow colour after days of storage. It is possible that this acid is
involved in colour formation mechanisms, during storage. Sugar produced by
carbonatation is more prone to form colour during storage than sugars produced by
sulfitation due to the inhibition of Maillard reaction due to the presence of sulfites

34
(Farber, 1975). However, the bond sulfite-carbonyl is not stable and SO2 may be liberated
with time and colour may return, as in plantation white sugar storage (Getaz, 1989)

6.5. Recovery
In Recovery section there is colour formation, due to Maillard reaction, in massecuites (in
boiling and crystallizers) and in molasses storage due to their higher concentration and
impurities present. This colour formation can happen with at high reaction velocity
causing explosions. Any storage recipient of massecuites and molasses must be ventilated
and with the security devices to minimize consequences of possible explosions.
Chromatogram and ELS diagram of molasses are presented in Figures 22 and 23 (Bento
et al, 1997). Sugar produced in Recovery section is returned to Affination. Colourants
integrated in this sugar will influence affination liquor quality. However, there is not
much information on colourants composition of these sugars.

140 250000

120
200000
100
Abs.x1000

80 150000
MicroVolts

60
100000
40

20 50000

0
0 10 20 30 40 50 60 70 80 0
Rt (min) 0 10 20 30 40 50 60 70 80 90
Rt (min)

Figure 22 - Chromatogram of Molasses Figure 23 - ELS diagram of Molasses

7. Conclusion
Study of sugar colourants in white sugar peocessing is important as colour is one of the
most important sugar quality parameter. The knowledge of colourants origin, formation,
destruction or removal during sugar processing is an important step to achieve the
mastering of sugar production and refing techniques. Identification of those colourants
that contribute more for final sugar colour, quantify their integration capacity into sugar
crystals and the necessity to control colourants returning to the beginning of process in
sweet waters and C sugar are important issues in sugar processing. Decolourization

35
systems are available nowadays but, as referred by M. J. Kort thirty years ago, attention
must be moved into other direction:
"Efforts would be better directed at the prevention of colour formation than at the
removal of the colourants" .

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AKCNOWLEDGEMENT
The author wishes to thank LEPAE / Faculdade de Engenharia / Universidade do Porto
- Portugal for their support.

38

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