•
Ití¡4""
UV spectrophotometric analysis of cañe sugar colorants
Analise espectrofotometrica UV de los colorantes del azúcar de caña
Luis San Miguel Bento
Sugar colourams and refmery sugar solutions are siudied using UV
specirophotomein m the range of 220 and 340 nm of wavelenghts
i'NUV - Mear L'llra \Vlet i. A relaüon beiween absorbancies at lov.
and high wavelenghts is calculated and, b> this means. a qualitahve
siud> of sugar colorams changes durmg cañe sugar refinmg procer
is performed. Coloran!;, changes in raw sugars. affination. carbona-
laiion, decolorizaúon and crystallization are estimated. A colorants
diagram throuch ihe refirme process. from raw sugar 10 whitc sug-
ar. is performed using this technique.
1 Introduction
Sugar colorants comprise a complex mixture of cnmpounds of differ-
eni origins and nature. Some colorants which orginate in sugar cañe
migraie to sugar juices through the sugar milis and refmenes. These
are cañe pigments. mainh fiavonoids and other phenolic com-
pound*. Other ¿ompound";. noi colored. also pass to factory and re-
finery sulutions becoming colored durmg the process. These are
color precursors such as invcrt sugar. phenolic acíds. nitrogen com-
pounds and othsrv Durtng the process other colorants are formed
such as caramcK. mcUnoidins. alkalme degradation producís of he.v
oses (ADP¡. ;md en/ymatic browmne producís as melanins.
Ultraviolet specira of these compounds and of sugar solutions of a
cañe sugar rofint-n. havc been studied. Visible and UV specira. alsc
called electrón spscira. repulí from encrgy absorption r>\*
(hat are excited from lower energy levéis lo higher ones. Most ab-
-.orption bands m these spectra are bread because each electronic en-
ct¿\! is associated wtih i t s longitudinal and rotational vibra-
lions. Electron trarrsition ma> occur from am of several \ibrational
and roiationai siaies of one electronif levcl U' anv ot several rotatton-
al and rotauonal Mates oí" a higher le'-el (Solumans. 1992). As a re-
sul!. UV spectra de nol presenl defined peaks. In this stud> ihe área
of the absorpnon obscrved betueen 220 and 27? nm and bemccr,
27? and 340 nm i.s calculated. Both áreas are referred 10 the unit\:
área of the ads(*rption at 273 nm mullipÜed bv the w a v e l e n g t h inler-'
••al These fractional valúes can gtve an idea of thecolorant composi-
lion of sugar ^ulutions A qiiaütative evalualion of coloran! composi-
üon through the l e t m i n g process i.s performed using this
- Equipment and methods
ln this study a GBC UV/VIS 916 Specirophotometer was used. Syn-
'tietic colorams were prepared accordmc to the literature (Shore et a!.
jyS4). Affination of ra^ cañe sugar was pcrformed b\g raw
sugar (98 gi wilh a sucrose solution at 70.2^ r d s f l O O g f in a rciating
closed flask immerscd ir a water bath ai 40 "C during 30 min. Sepa-
ration of affined sugar was camed out m a laboratory centnfugal
Solutions for UV spectra analysis of iugars and colorants were pre-
pared to give an absorbance of 0500 ± 0.100 ai 27? nm with pH
adjusied lo 9.0 i 0. 1 . Solutions were filiered ihrough a 1.2 ^m mem-
brane before UV readings. Distilled water was used as the blank so-
Los colorantes naturales del azúcar de caña y las solucionos de refi-
nana de azúcar son estudiados por espectrofoiomeiria de l.'ltro Vio-
leta a rangos de onda smre 220 a 340 mn. Se calcula una relación
entre el valor de la*, absorvancias a los ba\os tangos y los altos ran-
gos de onda. Por este medio es hecho un estudio cualitativo de las
alteraciones en los colorantes durante el proceiso de refinación. Son
avalladas las alteraciones de los colorantes en los azúcar crudo, afi-
nación, carbonatación, descoloración y cristalización. Utilizando
esta técnica so ha trazado un diagrama de los colóranles a lo largo
del proceso de refinación desde el azúcar crudo hasta el azúcar
blanco refinado.
5 Results and Discussion
Specira of caramel, melanoidin (alkaline). alkaline degradatior.
producís (f'rujioset anJ melanm are presenied in Figures I to4. Caf-
feic acid and chlorocenjc acid spectrü are presented in Figures 5 and
6.
UV specira of raw sugar and sugar produce- apparently do not give
much informaiion. Figure ~¡ shows the spectra of a raw sugar and
the respective affined sugar and affination syrup It is observed ihat
affined sugar gi\es higher abxorbance values thíin raw sugar. beiow
and above 27? nm. On the centran, svrup prc-sents a lower absorb-
ance in the vicinity of 273 nm
In order lo enhance ihis kmd of information. the valúes of L?.f (low
M,y\elen2th fractioni and H/.f (high wavelength fraction). dcfined bv
the followmg expressions. were calculaied:
Uf or HW = ( / Abs • dX) / Í(X, - ?.,) • (Abs at 273 nm))
The mlegral is calculated between the intervals Á, and X,. For the
calculation of LXf the valúes of A, and Á, are 220 nm and 273 nm.
respective)}'. For the calculauon of HXf. X, and Á, are 273 and 340
nm. respectivcly. Abs is the absorbance reading al ciifferent wave-
lencih.s. Fhe valué of 273 nm w.as chosen as u is Mtuated m a stable
zone of absórbante*, as can be seen from Figure 7. and n pues the
hesi resulis for ihi.s siudj. The valúes L/.f and H'\i represen! the frac-
tion of the áreas indijated m the example given in Figure 8.
3.1 Sugar colorants and related compounds
Uxing the meihod explained above. the l'V specira of synthetic .sugar
coioranis prepared m the laboraior\d other related compounds
were prepared. The valúes oí L/,f and H)J w-ere calculated CTable 1 )
In Figure 9, HXf \ersus L?.f valúes of the C(iloranis of Table 1. cañe
and beet raw sugar and syrups and fio\vcr cxiract prepared from red
carnauon petáis usmg acidified methanol are shown.
Four different ¿ones can be considered in this diagram. Caramel is a
ivpical compound of the zone were LÁf ar»d HXf are lower ihan 1.0
Chlorogenic acid is a compound from the ?one were both fraclions
are higher (han unily. Caffeic acid is a compound placed in the zone
were LXf is lower ihan 1.0 and HX.f is higher ihar unit>. All sugar
solutions and raw sugars are placed m the fourth ¿one. where LÁf is
higher ihan 1.0 and HXf is lower than that valué. Meianoidms. alka-
line degradaüon producís and meianins are also cumpounds found in
thi'. zone
7:

seo 3 Wavetengtfi m nm
Fig. I: UV and visible spectrum of caramel
Fie. 5: UV and visible spectrum of cat'feic acid

O MM 1
i.
«oo o WavWefigc- «t ivr

Fig. 2: L'V and visible >pecirum of melanuidin WO.I «O.» SM 5

Fig. 6: UV and visible spectrum of chlorogenic acid

•oe c Wav«lenglh

Fig. 3: L'V and vkible -.pectrum of ADP Fíg. 7: L'V spectrum of raw sugar ( R | , affined sugar ( A i and affina-
tion symps (S)

l.OH-J

273 340 Wavaiengtn n ntr

Fig. 4: L'V and visible spectrum of meianin Fig. 8: Example of LXf and HXf colculation

:o Zucktrwd. I20ttW¡l - /Wí

 LM and HW vduefcof sOTnejnlOfanl^ •J
H«J
ll»M 0.571
0.447 I.OSJ
0.337 0.4K2
1267 OJ05
0.705
I.J» l.r.8"»
0.815
0.482
0.927

0.66
2
H^ i.
Fi£, 10: LX-f and HU valúes oí a Zimbabwc raw and affincd sugar

•
1.5
•
I
04
c.*-

Císcrvf-
I
0.5 15 25 'l^S 1.8 l-«5 '..7 1.75 1.8 \85
LA! LX(
Fig 9: LÁf and HXf valúes of *ugar culorams and K-lated com- 11. Lí.f and H).f valúes ot Cuban ra* and affined sugar

? H Ka* n.c.n
CaiK raw sugars teceived reccnily ai RAR Refinenes were jlso
eva'iiiaieij Affmaiions of ilicse ra» «.ugar^ were perfomwd as df-
scribed carlier. in TaWo -. íta^'.ion valúes oHiained fron- inu rjw
sugan oí differsni nugins aje shown In Figures 10 and 11. [he frac-
lion valui-vof aZimbatwe and a Cuban rav. sugar. iKeír alñned ^u^-
«t-. ai>d affination ^ytup^ jrc '.hftwr. For a hetier :ompariMin. in Fig-
ure "I, ihr íraclion valúes r»f ra* iupars. with respective alfmaiinr.
sugdis are presemcd. eompared wiih ra* safan- *iih an arttitrarx 0.98
valué of 1 1 . 1 • Qá 1.1 1,15
It is obseíA'ed ihai ihe H/.f valúes nf affinet) sugars are al way í higher
Üian ilw unei of ra* lugjrs li ií also i^bwrved thai ceruin affined Fig. 12: líelaiive posiimn of uffirwd rclauvc to raw
iugars iia\ hijjhei LXf Ihan the rc^pecuvc ra* sugars, and . l.Oi \uluc
behave n iht'>ppir.ilc way. ll is inieresunf lo noit ihai
i^ rclaicd tu ra» sugat ottgín indicaling a particular kind ot sugat
cañe compound- or mili itchmilnc> o( that región. Aifinalion syrupi
conlain a higher pcrceniage of toloranti as faetón coloranis inai de - i Liquors
crease the H>.f
Classifyíng rjw "¿ugaj coloranis ihis way one can make a ¡.elcclion or A íiudy *as maüc of i^ie efíeci of ireaimcm wiih u numher uf dilles
mixture oí sugars 10 be protesscd in ordcr lo have a uniform npem- coi dccolorií.mg agenvs on carbonataban liquor. UV spccira beforc
iion in the refining process, (rom ihc decoloritaiton nnim uí view and afler n'eatmcnt were ira.td and L/.t and HXf valúes calculated
Reaciion condilions uscd were a\. carhonated liquor was
diluida < 111 i wiih disiillcd water A portion of 100 mi of (his solu-
Table 2 IMand HM\aluesof row sugar. affined suparandaffinaunn uon was mued wilh activated carbón \2 gV. granular carbón i5 gi:
i-yrup of differcni ongin?. «yrenic slrongly basic anión resm i5 g,): at-ryltc sirongly baste ani-
LXf HU Uí HW ón tesin If g], calcium hydroxide 15 gi and 2 mi hydrogen peroxide
Bnzil ^
solunon (B*) Samples were apiuied in a waler hath at 5U '-C for 30
ClltM
rninuleü. A sample nt un'reated carbonated liquor WAS mdintamed
. ~'*¿ 0.708 1.6:: 0712
mider '.h¡ same condinons to serve as a control. Ajier filtration.
1.819 0.737 1 806 0.750
AfTiMiioo «ynip 1.672 0.692 1.587 0.706
sugai Solutions were prepared as dc^cnbeó earliei :nr L'V s(*.-.
irometry analytis. Fratiinn results are presenieo m Tablf 3 and in
S*u Figure 13. The mixture oí caibonaied liqunr with limt waj, cnrrect-
Raw -ug»t i.*n (1.706 1.703 Ü.TW ed 10 pH 7.0 btfort cun> mg. out ihí L'V reading
Affined iugu 1598 0-768 Ii is observed thai H!) rtecoloriíing agents usíd produce »n increase in
Affinauun si-rup l.MS 1.718 ()(,«! LX.f fraciiotí niraning ihal ihe removed ci^lürann ha\ a lower L>J
Table 3: LXf and HXf valúes oí sugar solutions after difieren! HXr' and LXf valúes at ditferenr process stages of RAR
decolorization ireatmenis relinery
LXf HXf
Curbonsted L.iquor .244 0.721 Raw sugar .391 0"7
C.L. + Activated Carbón .274 0 6?7 Al'fined sugar .438 0.81
C.L. +• Granular Carhon .315 0 608 Affinalion syrup .326 07*
C.L. + Sl>rcmcResm .344 O.fi77 l.iquor írom retovery .335 08C
C L. + Acrvlic Rcsm .322 0.748 Sweet waters .351 0.71

C.L.*Ca(ÓHí. .183 0.630 Melt liquor 361 0.79

C.L. + H,O, .398 0.6<)S Carbonaied liquor .208 0.72
Dccoloriíed liquor .272 0.71
Conceniratcd liquor .283 0 7!
H/J
Feed syrup for pan .100 0.69
r\ . -
U. 76 Whue ^.ugar massecuite .134 0.69
^^ Acrylic resin
0.74 i Rctlft€Ú syrup 1.086 0.69
L quor ^^--*****^ Whue sugar 1.267 0.78
0,72 ?^~-
—
0.7 / \8""""-•^ / Vs.
^HjO,
^^-^^^
Styrenrc resm HXf
0.66 /
0.82
Liq. recov. Affined sugar
0.64 /
* Acfivated \2 1 Ca(OHl? ca*txín 0.8

-y
WNte sugar Me" \ liquor^
\Granularcartx>n
0,78
06
1.15 1-2 1.25 1.3 1.35 1.4 1
45 0.76
/ y^ S-^x Raw sugar

LXI / s ^
/ /
Aftinatton svrup
Fig. 13: LXf and HXf valúes of carbonated liquor before and after 0.74
treatment ^Carbonated tíquo '
O-72 Ref. / ^>T
syrups ^^^-^ Dec./ Conc liquor
0.7 ' . -r-^-
valué Massecuite
anís with higher HXf. lowermg this valué in Ihe final produci. As 0,68
iicrylic resins have a lower capatity lo adsorb colnrants this et'feci is 1.1 1,2 1,3 1.4 1.5
L/.f
not ohserved and H>,f is higher rhan for other decolonzers. The ac-
Fig. 14 LXf and HXf valúes dunng refinmg prcte.ss
tion of lime on carbonated üquor produces a deviación in the direc-
iion of alkaline deeradaiion produc: valué (1.064. 0.571) indicniins
the t'ormaiion of thcse kind^ ni compounds

3.4 Colorants' balance through the refining process

All the rcfinen. sugar wlutions from raw sugar to while sugar were
studied al Ihe RAR refmer). ¡akin¿ average surupíes ofeach produce
al 15 minutes intervals d u n n a ^ne hour Sample.s are laken accordine
10 rcsidence tune al each siage. Resuiti of LÁt and HXf are presenied
in Tatile 4 and ihe balance diagram presented in Figure 1-1. The uriñe
sugar reading \vas made in a •iO'T- rds solutiort.
It is observed idaí from affined sugar to *hiie iugar massecuite, the
fraction valúes cross the diagrum from the lop nghi position to the
boiiom left This means ihat dunng refming. at the same tune ihai
polypheríol colorants with a high HXf are removed, there is a forma-
(ion of faciory colornnts with low HÁf and low I.Xf valúes. Using
these diagram vaiues. one can evalúate the behaviour of colorants
dunng the refining process.
Affined sugar is mixed with sug.ir l'rom recovery and sweet waters to
produce melt liquor. Mn\i:ment towards the bottom left of the melt
liquor position in thc diagram indícales possible caramclisation oc-
cumng dunng dissolving.
Carhonatation is a good process lo purify melt liquors. A decolon-
/.ation of around 60fí is obtamed. Dunng this process. alkalinilv in-
creases to high levéis and destrucción of reducing ^ugars can occur.
The movement of carbonated liquor in the diagram in (he direclion oí'
alkalint degradation product colorants indícate^ ihis possibility.
Afíer carbonataiion, the liquor is decolorized through styrenic ion
exchange resins. Movemenl obseded m this diagram is in agreemenl
with ihai observed ai Figure 13. At'ter the e vapora tors, liquor is
mixed with refined syrup in ihc v.hile sugar massecuite. The changes
seen in the diagram indícate Ihe formalion of colorants during crys-
ialli¿alion. possibly caramels. After centrifugaron, white sugar is
separaied from the resultinc refmed syrup The while sugar position
m ihe diagram indícales the presence of compounds ¡n the sugar with
high HXf valúes.
4 Conclusión
UV spectra coniain much more informanon (han ihat obiamed by
comparmg only spectra máxima of each compound Companng ihe
absorbance behaviour at low and high wavelcngths in ihe UV spcc-
tra. one can have an idea of ihe behaviour of colorants d u n n g the
refining process. More information is obtamed by comparing specira
valúes afier a process step rcmoval of coloranls m decolorizaiion.
separanon of syrups in affination and wiiiie sugar masseciiile centrií'-
ugation. addition of lime. etc. This process uses simple equiprrtím
and dt'Cs noi need spccial iraming uf personnel for its manipulation.
In 15 minutes, the valúes can be determined and calculated and a
complete diagram of colorants through ihe refinery can be obtained
in half a shift This process can be useiul m make a rational mixture
of differem ra\>. sugars lo refine. evalúate any change produced ¡n the
refining procesi, and dctect any anomalous coloran t formation in the
process.
Acknowledgment
The author wishes to thank the help of his DID colleagues. specially
Sitiaría Sd. dunng the preparation of this paper.
Bibliography
Shore. M : Brout¡hti>n, N.W.: Duilon. J.V.- Sissotts. A.: Factors afTcciing
v-hue sugar coluur. Sugar Tiíchnolog) Revicws 12 {19841 1-99
Sohmoní. T.W.C. (1992): Organic Chemistr>-, 5ih editmn, John Wilcy &
Sons
Author's address: Luis San Miguel Bemo. RAR Refinarias de Acucar
Reunidas. S.A.. Apt. 1067. P-4101 Porto Codex, Portugal.; E-maiI:
Ibento&iclepac.pt.

I
 I
Troc . o 4

SIT paper n" 683

Appliíation of I V Speetuphotoinetry to study sugar colourants

the rcTmmjj proccss

Luis S:i" Migui'! liento

R.AR - Kcfmanas de Acucar Reunidas, SA
Oporto - Portugal
E-mail: lbento@telepac.pt

1NTRODUCTION

The iludy of sugai colouranu» 111 taw sugars and in refimiig producís has been one of ihe niain
pnorities for sugar technologists A yreal efTon has been niade in the last few years concerning
the sludy of colourants composition in ditTerent step* of sugar refmimj, using sophisticated
techmques These techniques use HPLC (Patón et al.. 1982, 1992), GPC (GodshalU992;
Lincear et a!., 1978), TLC (Lincear el ai, 1978), ¿visorbent resins (Chou and Rizzuto, 1972;
Godshall and Roberts, 1982) and dinlysis (Robens and Godshall, 1978) All these isclinKíucs
are of the utmost importance lo identify and characlense sugar colouraiiis composmon

For daily work m faetones and refmenes, sugar lechnologisis neeü a practica!, rapid and simpí*
leclinique to gtve them, m the shortest time, informatton concerrang colourants con^wsition in
sugar process.

Sugar colour measured by Ihe absorbancy at 420nm (1CUMSA, 1994) has been used ip
evalúate colourants in techntcal sugar solutions. Other simple tests have been introducid tu
give more infornution concerning colourants nature (Clarke L-I ni. l<-)84). One uset'ui
parameier is íhe indicator valué, IV. This is defmed as she quotiem of llie absorbancy ai -4?.0tini
at pH 9,0 ajid ai pH 4,0 It gives an idea of the colourants naiure as absorbancy varíes with pH.
depending on the coicurajits con^josiíion (Clarke. i l >87)

In this paper we presetn a (echnique Ihaí can be a ijood tool both for coiourants identtficntion
and forsuidy of their bthaviour in each refimng siep The lechmque, described earlier (Bemo.
1995), consists of iracing the specttum of Ihe sugar solution, between 220 and 340 nm Tttts
raiige of ratita t ion corresponds lo a UV spectrum zone named NUV - Ncar t'itra Violet or
quart/ UV Aosorbancy ot" electroniagnelic radialion in thts /one of the spectrum corresponds
to electrontc irnnsitions between nwlecular orbual \\uhin the moléculas Watei. sucrose ;uut
other carhobyrlratfre do not absorb electromagn*fic radiation tu this zone of the spectnim
Natura) col curan ts usuaJly possess double oonds. with different degrees or conjugation m theu
structure Thcso bonds sjive rise to n -* u* transitions that absorb cíectromagnetic radiation in
the UV región The iransmon energv depends on the conjugal ion deyree, decieasmg wnh it,
the elecuonic environment within the molecules and with the solvem usad Other tran&iiion
lypes, such is ihe n -» n* type, can occur wdh radiación in the NUV xone (C'reswefl. 1972)
Usualry the UV spcctra presen! broad bands» due to the existence of vtbratíonal and roiational
levéis associatcd with each electronic energ> level (Soloman, \W2) The widih ofthe baiid is
aíso. m part, due lo instrumental ion (Creswdt, 1972)

The spectra correspondiny 10 caramel, rnelanoidme, aikaiine deyradanon producís, chloro»enic

and caffeic acids are presemed m figures 1. 2, 3, -i and 5 respectivety The NUV spcctraofa
raw sugar wtth the respective affined sugar and syrup is presented m ft«ure 6

In ihe technique descnbed riere, the absorbancy in the reipoii of 220 and 273nm is compared
•A-ilh that of 273 and 340nm, both related to ihe absorbancy al 273nni Tríese pataineiers g»ve
a.i :-dea ot" the colouranis cotnposition of the difieren! sugar solulions throughout the refining
Dr->cess The method was lested with niixtures of known orgafiic compounds and wilt)
technical sugar soiutionü Coniposition of ihe coiourants frorn raw sugar. affínation,
carbonaiaíLOn, decí^oufizaiion and crystalhsation was evaiuated Spccial attention was paid 10
ton exchange resin decolourization

MATERIALS AND METHODS

In thjs study a GBC L.V7VJS 916 Specnophoionwier was used Iniegranuu \\as tained oui
usmg the GBC software Symheíic colouranís were preparad according to the lileralure íShore
el ai. (984)

Raw cañe sugar affination was carried out by nwxing 98g of raw sugar wilh ÍOÜg of sucrosa
solunon at 70.2° Bn\ m a ¡otatiny closed tlask mmiersed m a water bath al 40*C. for 30
minutes Separation was done in a Laboratory cenlnfugc withou! washmg

Solutions for UV Spíctra anaJysts of sugars and coíourants were preparad so as lo grve an
absorbancy of 0,500 ± 0,100 al 273nm with rhe pH adjusted to 9,0 ± 0,1, Solutions were
filíered ihrough a l.2u.m memtxane before UV readings were taUn. Later the absoibaiic>
valué was changed lo 0,700 ± 0,100. Foi consisient resulls absorbanaes ai 273nm rmist be
between 0,300 and 0.800 DistiHed water was used as a biank solunon

Valúes of LXf {Low Wave(e"yth ftactton) and HXf iHiyh Wavelongth frnction) dcfincd bv- the
follownuj expressions, wereevaJuated as
 LXf or HXf =* (J Abs x dX) / ((AbSo) x (X2 - X,)) 11)

The integral is cvaluated. between the limits X} and X 2 For the calculation of LXf tbe vaJues of
X, and X3 were 220nm and 273mn, respectively For the calculation of HXf, X, and X> wtrpí
273 and 3-IOnm, respectively Abs represents the absorbancy readings at dirTerem wavelenyths
Abs3 represents the absorbancy at 273nm. As can be demonsrr.ited, usmg the Beer-Lanibert
la\ ihese fractlons represen! the quotient given by the expression

le^í/Ie0, f; (2J

ln the precedmg eíjualion, em¡ , represents the averaíje of fhe molar absoibancies. :;„ af
compound i, with a molar fraction £, m the considered spsctrum /.one, given by.

e°, represents the molar absorbancy of compound ¡ al 273nm

RESULTS AND D1SCUSSION

Mixtures ofoi'ganic cumpoutuls

ln oiíier to íest this method of evaJuating mixtures of cornpouuds we caJculated and measured
Ihe valúes of absorbanoes of two organic compound mixtures, catechoi and sinapic acid.
Resulta are presentad m Tahle I and Fnjiire 7

Table I - Mixture of Catecho! and Sinapic acid

Calechoi % Sinapic Acid % LXf ' l.Xf Hxr HXf

Experimental CaJculated Experimental Calcula! ed
100 - 0,777 . 0,2S1 .
80 20 1,120^ L1I7 0,853 0,905
60 40 1,326 1,312 1,247 1,264
40 60 1,435 1,440 1,466 1,498
20 80 1,499 1,529 1,634 1,662
- 100 i,W - 1.7X3

Calculatíons were made by usiny Ihe Beer-Lamben Inw and ustng the molar absorbanoes
determined experinientalty. As obscrved, there is nol a significanr devintion beiween rhe
cnJculaíed and the experimentaí valúes

ln another test, solutioiis of CafTeic Acid, Sinapic Acid, Catechoi and Caiechm Hydraie were
prepared as descnbed The NUV spectra was traced and fractions l./.f and HXfwere calculaied
accoiding to equation (I) A mixture of these compounds at concenirations wnh similar

'- ;
 i'f vV $*«•# ««MWíT.víry ¿í' -ííí*'*ír í-'-'í^ iVíi>;;fv;??.'i írn-üa^u! fiK^-jya-üLt:^" ¡S-f'.'J"., íW.yj

absorbancies weie prepared and the sptxtrum and fraction valúes caicuiated Valúes are
presenied in Table 2 and m Figure 8

Tíible 1 - M I M U T of CafTe'.c ACKÍ. ratcichm Hyúraie, Catechol and Sinapic Acid

Abs al 273 nm LXf HXf

Caffeic Acid 0,6424 0,939 5.148

Catechin Hvdrale OJ089 2,419 0,440

Catechol 0,7645 0,760 0,278
Sinapic Acid 0,6592 ¡,575 1,758

Mixture 0,6750 1,489 0,891

Averaye ("i Í,41l> O.Ü72

(*) Average of \f valúes weighed by litó absorbancies at 273nm.

The results in Table 2 show a good agreement between the computad average and the
experimental valué for the mixture

Sugar cnloui-ants ¡ind relnted conipountU

Using the method explained earlier, NL'V spectra of synthetic sugar coíourants and related
compounds were traced. Valúes of l.Xf and tlí.f were caicuiated and are presented in Table 3

Table 3 - Symlwtic coloura-ns and relnted compounds

LXf HXf

Alkaline degradation producís 1,064 0,571

Caramel 0,337 0,482

Chlorogenic Acid 1,336 [.689

p - Couniaiic Acid 0,4% 0,815

Melanine 1.795 0,482

Melanoidine (aikaline) 1.202 0,927

 ÍS./.T-»

!n Figure 9 is repiesemed LXf versus HXf of ihese compounds. It is also presentad in ibis
Figure cañe and beet raw sugars and syrups and a white sugar

Four difTefent zones can be comidered in this diagram. Caramel is a typicaJ compound of Ihe
zone where LXf and HÁJ'aie lovver than I,U. Chloroyomc actd is n compound from the ¿one
where both fracttons are higher than imity CafFeic acid is a compound placed in the xone were
LXf is lower than 1,0 and HXf is higher than iffüty All sugar solutions and raw suyars are
placed in the fourth ?-one were LXf is higher than 1,0 and HXf is iower than that vaiue
Meiajioidmes. ADP aJid Melanineí are compoundí ¡n ilus ?one

In this study we have evalualed the NUV spectra of cañe raw suyars received recentiy ai RAR
Refmeries Affination of these raw sugars were performed as described earlier.

ID Tcible 4 is presenred fraction vaJues obiained with four raw sugnrs of differe-nt ongins !n
Figures ¡O and 11 are presented the fracuon valúes of raw suyars from Swaziland and Cuba.
respectwely It is interesting lo note that for the case of the first raw sugar, the positions in Ihe
diagram UXf vs. HXf of cañe syrup, raw sufiar, afllned sugar and white sugar, in this order, are
approximñteíy in a line poinling ín *he directton of higher HXf and !ower LA.f. This indícales
that the coiourants wrth these charactenstics aie the ones with more affinity to the sucrose
crystaJs.

For a bctter comparison, fraction valúes of raw sugars with respective affmation sugars are
presented, considenny raw sugars with an arbilrary vaiue of(l,0 . 1.0), in Fiyure 12

l'able 4 - Raw sutiars

Bnuil Cuba

U.Í H^.f txr Hxr

Raw Sugar 1, 712 0,710 1,622 0,712

Affmed Sugar 1,796 0.736 1,806 0,750

Affmalion Sytup 1,643 0,685 1.587 0,706

Swa/ilaiid Zimbabwe

LXf HX.f LXf iar

Raw Sugar 1,627 0,706 1,703 0,709

AtTmed Suyar 1,579 0,789 1,598 0,76»

AfTtnation Syrup 1.645 0,672 1.718 O.bSI

-••
 It ts observcd thal all affined sugais have higher HXf valúes than those of raw sugars U is also
observed ihat certam afñned sugars have higher LXf than the related raw sugars, and others
bchave in the opposite way U is intercstmy to note Ihat this tendency ¡s related to raw suyar
ongin. indicating a paiucuhu kmd of silgar cañe compounds or mili technology used at that
reyton AfTinauon syrups contain a higher percentaje of colourants that tiecrease the HX.f,
corresponding to colourants with a low lome charge, as it will be later reterred to

A study was made using carbonated liquor reactmy with different chemicals The NUV spectra
before and after treatment was traced and LXf and HXf valúes calculated Reaction conditions
used were as follows carbonated liquor was diluted (I !) with distilled water. A portion of
lOOm! of this soiuhon was mixed with activated caibon (2y), yianular carbón (5g), st>Tenic
strong base anionic resin in chioride form Í5»l, acwhc stronij base aiiionic resin ¡n chloridf*
form (5g); calcium hydroxide (5g) and hydroyen peroxide 3% v/v (2rrf), Samples were
agitated in a water bath al 50T for 30 minutes. A sampíe of carbonated liquor was maintamed
at the same conditions. After filtration, sugar solulions were prepared as descnbed eailier foi
UV spectrometry anaJysis Fraction results are presented in Table 5 and in Figure 13 The
mixiure of carbonated iiquor and lime was corrected to pH 7,0 before NL'V rending.

TaWe 5 - Reactions with Carbonated Liauor

KXf HXf

Carbonated Liquor (C L ) 1,244 0,721

C.L + Activated Carbón 1,274 0.637

C.L. + Granular Carbón V 1.315 0,608

C L + Stytenic Restn 1,344 0,677

C.L -i- Acrylic Resin 1,322 0,748

C.L. +Ca(OH)j U «3 0,630

C L ^ H,0, 1.398 0,698

It ¡s observed thal all decolouri/ers empioyed pioduce an mercase orí L>.f fraction. ihis
meaniny that the removed colourants have a lower LXf valué As white sugar also has a lower
lAf valué, this indícales that sugar decoloiin/alion agents are renwvmg the colouraiits that
have tiwre afTiniiy to the white sugar, as it was expccted It is also observed that adsorben!

214
decolounzers adsorb colourants with higher HXf lowering ihis valué m the final product The
behavior of acrylic resin will be explnined laier. The action of lime on carbonatad liquor
produces a deviation m the direction of the ADP valué (1,064 , 0,571 j indicaum» the foitnaüofi
of this kind of compounds when lime is added lo the liquor

t'olimniíiis (hruiiyhmii decolourizarion

in order to study the behavior of sugar colourants throughout ion exchanije resins we used a
colimm of one liter of resin and a column of 70ml of resin. The styrenic resm used was IRA
900C m the CI" form Carbonated hquor ÍCI.) from ihe refinery was used to be decolounzed
by the resm.

Reyer.eranori WOE performed in such a way so as to remo ve aJl the colourants m the resm.
These colourants compnse i) coiourants adsorbed to resin matrix due lo hydrophobíc ínter
actions and removed at k>w salí concentrations (AD); li) colouraní? attached !P the resin due
to ion-exchant»e rneclianisnis and removed al hitjh salt concenírntions (EXJ; iii) colourants Iliat
are noí removed by NaCI regeneraron and remam fixed to the resin (FIX)

In the one litei column, after 40 BV (bed volume) of diluled carbonaied liquor (3'¡ of w.iter)
in each decolounzing cycle. regencraiion was nnde m thiee pans usmg one BV of NaCI at
30g/l to remo ve AD colourants, three BV of N'aCI at lOGiy'l to reniove EX coiourants; and a
mixlute of equai parts of Ethyl alcohol, Acetonitrile, NaCI 2M and NaOH 1M to remove the
FIX colouraiilb,

Results of decolourization and revjeneration are presented in Figure 14. Valúes oflAf and HXf
cf effluems, during re«eneration are presentedSn Figures 15 and 16 It is observad that HXf
¡nrrenses with salt concent rallón, indicating thai compounds with higher anionic charge,
released ai riigh salt concentrations (Sentó, 1989), have hiyher 1 IXf valúes.

Dunng the decolounzation cycle it is observed thal LXf valúes of decolounzed liquor
decreased This can indícate that during the cycle, resin lose the capaciry to reniove AD
colourants and ar the end of the cycle onty EX and FIX colourants are removed

To study and make a balance of colouiant gioups released dunng reyeneranon. \vc used ,1
70ml column wilh a styrenic resm (IRA 'XJOO tn the Cl" fomi Carboiules Liquor was diluted
at M and 2800ml were passed through the column. At the end of the cycle, liquor in the
column was disiocated with distilled water nli a total volume of decoiounzed hquor of 3000ml
was achieved. A volume of 280ml of feed liquor (diluted at I I ) WAS niade up with distilled
\vaier till 300ml and absorbancy at 273nm and at pH 9,0 was determmed

'-'

á
Regeneration was then inade m difFerent steps as follows.

- 400m! of NaCl 0,2M (to remove AD1 colcurants)

- 400ml of NaCl 0.4M (to remove AD2 colouianis)

-400mIofNaClO,6M (to remove HX! colotirants)

- 400inl of NaCl 2.0M (lo remove EX2 colouranisl

- 400ml of equal parís of Ethyl aJcohoi; Aceíonitnle; NaCl 3M, NaOH iM

fto remove F1X cctourants).

After each regeneration solution was passed throughoul Ihe resin. Distüled water was ihen
passed ro renwve the chemicaJ solution, iilt a total volurr.e of 500ml was reached m each
fraction Absórbanles ai 273nm and at pH 9,0 were measured at each fraction.

Results are presentad in Figure 11 and Table 6.

Table 6 - Decolounzation with resins

Vg|.(l) AM273) Di). Tol;il LU AMLX) ii?, r AbifHM

F.L. 3,000 0,6519 1,667 3,260 1,335 4,352 0,661 2,155

ADI 0,500 0,8541 5 2,135 1,143 2,440 0,521 1,112

AD2 0,500 0,7541 4 1,508 1 .533 2,312 0,591 0.891

EX1 0,500 0,7995 2 0,800 1,168 0.934 0,746 0.597

EX2 0,500 0,6911 2,5 0,864 1,169 1,010 0,829 0,716

FIX 0,500 0,7765 10 3,882 1,167 4,530 0,812 3,152

Tolal I2.44S ;,2M 15,578 0.693 8,623

C.L. 3,000 0,8332 5 12.498 1,236 ¡5,448 0,692 8,649

Dtff. % 0.39 1,20 0.83 0,14 0.30

The sum of the valúes of Ihe fractions of the effluents duriny re^enetation, wuh the valué of
Ihe fine liquor (F.L.), would be the saine as ihe Liquor emenny rhe coiurnn (C.L.), if all the
colouranis were removed from the coluirm Tlus can be observed in the lasi iows in Table 6
From these resulls we can concltide thai practicaily all the colourar.ls were removed from res»)
wuh this regeneration In Table 6. Abs(LA) and Abs(HA) represents Ihe médium total
absorbancy (Absorbancy x volume x diíution facior) in Ihe low or hiyh wavelenglh región Dil.
in the table represents the dilution factor.
Tne lower capacity of removal of FIX colourants by acrylic re&ins can explajn the behavior of
these resins, observed earlier (Figure 13)

Colourants lliifmy;tKn>t the rrtimnj'. Proccss

Tm? study was cained out ai RAR Rcfmery by taking average samples of each product at 15
minutes Ínter vals dunng one hour, aJIowing für the residence time at each stage Results of LXf
and H>.f are presentad m Table 7 aiid the baiance d¡agram is shown m Figure 19 The \vhite
sugar readtng was made m a 40* Bnx solution

Table 7 - Colourants through the Rp.finmij Process

LXf HXf

Raw Sugar 1,391 0,773

AÍTined Sugar 1,438 0,806

Affination Syrup 1,326 0,751

Liquor from Recovéis 1,335 0,805

Sweei Waters 1,351 0,774

\1etf l.íquor 1,361 0,79!

Carbonated Liquoi 1.208 0,727

Decolounzed Lwjuor 1,272 0,715

Concentratecl Ltquor I 7 283 0.710

Feed Syrup for Pan 1,100 0,697

Milite Suya/ Massecuite 1,134 0,697

i

Refined Syrup 1,086 0.699

\Vhite Si.iyar 1,267 0,786

It can be observed ihat, from aíTined sugar lo whue sugar massecuite, fractton valúes cross the
diayram froni the top nyht position to the boTtom ¡etl

AÍTined suyar is niixed with suyar from recoven,- and sweet waters to produce nielí liquor
Dislocaiion to the bottom íeft of the nielt liquor position m the diaijram indícales the possiWe
colour tbrnution during nielt.
Carbonata! ion is a good process to punfy mdt Iiquofs. A decolourizatton around 60% is
obtawied Durtng this process alkalinity increases to hisjh levéis and destruction of reduong
sugars can occur Deviauon of carbonaled liquor in the díagram in the direction of ADP
colouranls, indícales this possibtljty.

After carbonatation the liquor is decofoufized ihrough styremc strong bnse anionic tesnis The
behavior during decolourization was ariaJyzed earlier

After íhe evaporators the liquor is mixed with refined syrup m (he white sugar massecmte. The
deviation shown m (he diagram indicaies possjble carameli¿ation m the pají After
cenlrifuijation, wtíite sugar G separated from Ihe resultint; refined syrup. The white sugar
posnion m rhe diagram indícales the presence ofcompounds m íhe sugar with high HXf vaJues

The same kmd of diagram of colouranls throuijhout the refineiy, m a difieren! occasion, was
traced making the specira measuren-.ent at pH 9,0 and 4,0 Resulls are presented m Figure 18.
It is observed (hat there is a greater dcvialion from the valúes at pH 9.0 ín the white end of the
refinery

Tliis diatjram was píotted every oiher week ai R AR Refinery Resuits of the fatest balancea are
presented m Figure 20 It can be observed that, although startiny with raw sugars of different
onguis, the resultmg fine liquor has a similar compositíon

roloiirnnts c va) 11.1 tío u during Refining

Usualh/, colourants dunng íhe refimng process are evaluated as ICUVISA colour, which is
measured al 420nm As Dr Frank Carpenter referred to {Farber and Carpenter, 1972) colour is
a liiree d.mensicrul páramele: Only one parameter does not rive A complete inforntaíion of
the kind ofcompounds presen! For evaJuatiSn of suijar coinposilion, throughout íhe factory oí
refininy process, a system corisistiny of three parameters, will be more appropriate. One
paranKter will indícate tlie coiourants intensity, whilst the other two, íhe quality of the
colourants The colour measured at UV will be referred to as UVC and is defined as
UVC-A(Uf;HXf)

in whjch A is the Abs(UV) referred to íhe unity of solution Dry Substance and km of cell
length:

C » Abs(UV) x (IOO/ Brix x density x celi length) x 100

AbsíUV) = Absí273>» (LXfx 53 <- HXfx 67) / 120

As an example comparisons are made in Table 8 of the UVC valúes of two raw sugars wtth
ideniical ICt'MSA coiour bul differem colourants composilion

A ¡00
 Table 8 - UVColour

UVColour ICUMSA Colour

Raw Silgar 1 3855 (1,74; 0,72) 2660

Raw Sugar 2 3636 (1,57 ; 0,73) 2625

By using this process of colourants measuremenl, more complete mformation will be oblained
For white suyar, where a visual appearance of sugar colour is requested, this sy^tem of
coiourants measurement will not be adequaie

l'.rrors and ¡ufliience of lirix

A test was niade usiny a techmcal s¡js;ar soiunon at difierent dry substanees and at different
colourants concentraiion This was achieved by adding diííerent quantities of waler and
sucrose (50:50) to the carbonated liquor solution. Results are presenled in Table 9.

Table 9 - Imluence of Bi

LXf r>iir. % HXf Diff. %

L(.a;KrBrix 1,279 0 0.727 0,55

LO<cl5°Bri\x 1,274 0,39 0,726 0,68

1,284 0,39 0,737 0,82

LC>Suca]IU-B\x 1,285 0,47 0,731 0

1,274 0,39 0 ; 74i 1,37

LC*Suoff>ÍO*8i 1,279 0 0,720 0,68

AvcrjRC 1,279 - 0.731 -

As can be observad the mfluence of sucrose concentraron and the colourants dilution do not
stgniñcantly afTect the resutts
 Conclusión

By eonipaiing ifoe absorbancy behavioi al lo* aiid lugh wavelengíhs in the N U V spectra we
car: have an idea of the colourants behavior durmg the refinjm; process More mformation is
obtamed by comparing spectra valúes after a process step removal of colourants m
decolourization, separation of syrups in affination and white sugar niassecutte centrifugación,
addition of lime. etc. The process descnbed uses simple equipment and does not tequire
speciah¿ed peisonnel In 15 minuies the valúes can be determinad and calculated and a
complete diagram of colourants throughoui the refineiy can be obtained in half a shif! This
process can be useriil to make a rational mixture of differení raw sugars to refine, evalúale any
change produced in the refiniíig process and tisieci ¿uiy .inomalous colourants formation in the
process Usmg a three parameter system to define colourants composition in the refinery can
be very helpfuJ for Ihe monitonng of the refiníng operation.

Acknowledgment

The author wishes to thank the help of his DID colleagues, speciaJIy Susana Sá, during the
reafization of this paper. Tliis paper is part oí*a EUREKA Proj<a financed by P E D I R Program
and STRIDE Program (resins). Help from the Colteagues of Minho, Oporto and Coimera
Umversity and IBET were appreciated.

Bibl¡ogi-;iphv

Bento L.S.M., 1989, Sugar decolourization by ion exchange resins with regenerant recovery,
Proceedings of S1T Conferwice, 1 76-^)0

Bento L S M.. l<>95, UV Spectrophotometric analysis of cañe sugar colourants,

Zuckerindustnc (m press)

Chou C C , A. E. Rizzuto. 1972. Tne acidic nature of sugar colorants. Procsedings of the
Technical Session on Cañe Sugar Refining Research. 8-22

Clarke M.A.. R.S Blanco, M.A.Godshall, 1984, Color lests and other indicators of raw sutjar
characíertstics. Proceedtnysof SPR1 Conference. 284-302

Clarke M A , 1987, Cañe sugar colorants in processmg operations, Proceedmgs of CITS

Conferetice.67-81

Cres^ell f.J , Runqutst O , Campbell M M., 1072, Spectral Analysis of Organic Compoimds.
Burgess Publishing Company. Minneapolis. Vltnrv

"•'•-
 £."A-»í.-.;;:V.''Vrí i»*" U\' Sj'wí.'ffpíiíí.v.'íVíyy V- flujy &¡í%¿r ivírtwwíí/i ÍÍ.'"ÍÍÍÍ¡JÍÍÍ¡L'/ f/«í/ígjí»rsí:c

Farber L , F.G.Carpenter, I972t Plant pigments as colorants in cañe susar, Proceedmgs of

Technical Session on Cañe Suyar Refining Research, 23-3 I
Godihall M A., EJ.Roberts, 1982, Phenohcs in sugar producís: their role in flavor and color
production, Proceedmgs of SPR1 Conference, 47-72
[CUMSA - International Commission for Uniform Methods of Sugar Analysis. 19-.M, MtMhods
Book
Lincear D , N.H Patón, P.Snuth, 1978, Techniques for ihe isolation of cañe sugar colorants,
Proceedings of TSCSRRP, 81-91
Palón N.H., P Smith, 1982, An HPLC study of the chan»es in coloranl composition foliowng
factory decolonsalion of raw liquors wiíh bone char, resm and gratuilai carbón,
Proceedinys o f S P R I Conference, 1-18
Patón N.H., !992, Sugar cañe phenotics and firsi express juice coloui, Intemationai Sugar
Journal, vol. 94, 1121, 99-102; 1) 23, 157-160; 1124, 170-176, 1125, 205-2! O
Roberts E.J , Godshall M . A , Some observaron orí the high molecular weight colorants in
sugar, Proceedmgs of the Technica! Session on Cañe Sugar Refining Research, 68-80
Sobmons T.W.G., 1992, Organic Chemistry, 5th edition, John Wiley & Sons
Shore M, N.W.Broughton, J.V Dutton, A Sissons, 1934, Factors affeciing white stigaf colour,
Sugar Technology Reviews, vol 12, 1-99
Williams J.C., C.L. Bhardwaj, 1988, The use of HPLC colour analysis to investígate (he
mechanismof resin decolorization, SPRI, 1-10

>*•*
. '
 SKíWíí.v/rv íi- *í:.'¿? tuar iVí;í«r.-i?í.'i &.-£.-u¿& ¡S-'-T-,

B.8BB~

H.Í.HH:

B.4H

e.ZB8-

e.efle 1
—j—
ZtJU.B -489.a ¿BH.B

Figure I - Spectrum of Caramel

8.688-j

8 . 4»

8.880

268.9 488.6 68B.8

Figure 2 - Spectrum of Melanoidme.

1 .888

8.588:

888

188.3 t>88.8

Fiyure 3 - Spectrum of alkaluie deuradatiün producís

 —t—
Z8B.B 4ftfi.fi MU! ti

Figure A - Spectrum ofChlorogemc Acid

8.888;

B.f.8R

tí.

8.

BHB_

208.0 498. B &8fl.ft

(nía)

Figure 5 - Spectnim of Caffeic Acid

fibsorbance
8. 889 i R Raw sugar
A Affmedstiijiíi
íí . ftfTinalion syrup

B.Í.HB

z-ia.e Z&8.9 2C8.9 369. M

v i T i i ' m p r (uní

Fiyure ó - Spectrum of a Raw Sugar

' .:
 H/.r '

:
.:

.
-•
1 1.2 1.4 1,6 1.9
LXf

Fig. 7 - Mixture of two compoimds Fig. 8 - Mixture of four compounds

u >. r

ttr.1S
0.5
• ftMI «««*

.
.•
3,66
O 0.6~ 1 1.8 2,5 1,56 1.64
LJ :

Fig. 9 - Synlhetic colourants and related compounds. Fig 10 - Affination of a Raw Sugar (Swaziland)

l
 '

Hi.f 1,12
•
-• 1.1 ri
.:

1.08 f

:. 1.06

1,04

1.02 !
o.7i r
A. Syup
1.56 1,6 1,65 1,7 1.7S 1.8 1.86 0.98 E
0,9 C.95 1 1,05 1,1 1,15
• Fig I I - Affination of a Raw Sugar (Cuba). Fig. Í2 - Relative position of Raw and Aííined Sugar of difieren! origins.

H?.f
0,76 0,9

08

ti?

J6

0.5

.3
0.4 !
• 1.15 1.2 1.25 1,3 1.35 1.4 1.45 1 1.1 1,2 1.3 1.4 1,5 1.8 1.7
l U Uf

Fig 13- Action of different agents on Carborialed Liquor. Fig. U - Decolourization with ¡on-exchaíiye resins.
Liquours and leyenerotion effliients
„. LU NaC 9» HXf
!£• . ^or\U
tr
i7 * 1 I-'
••y- 1

6 - , ^..--•*-
100
»-. ir
o.»
l-<
i-.'. ,' r
ÍS> 1.S - — f ._r
• 80 0,6 [
s
s> V4 ^ 4 Á í
r -1 • 6C 0, f
§
1 3 r / • 'V
fe; i . S
&
;¿
í \2 - / ;
*•"*""*"•••-•.
S-»^4^
40
^•v*
o.e r
V. L--**
2C 0.5 f-
&vi Vl;..t,r./-;
•
f *
1 , •'
.1-, . , '" ^
*Ví 1 ...1 ' n
U 0,4 -
A e • 4 e o 4C « «e 0.5
t 1 1.5 2 2.6 3 3.5
^ BV BV

Fig. 15 - LWduring retín leijeneration. Fig. 16-HXf duríng rain regeneraron.

H>.f nn
08.5 r—
EX2 085 ~--'

-- *N4¿

0,75

0.7 '

O.S5 "

06

0.55
1.1 1.1 1.3 1,4 1.5 1.6
Uf L>.f

Fig. 1 7 - Oecolourízaiion with lon-exchange retins (column of 70 mi). F>S- '8 - Colourams througliout the Refining Process at pH 4,0 and 9,0.

18
 0,82

«:
Í.KJ R*«W

'.•.•MÍ- 3utf* • Vrt uigi.pt

0,78

0,76

0,74
-
0,72

0.7 r,rt -j, ut^

0,68
1,1 1.2 1,3 1 t
I. >. i
Fig 10 - Colourants íhroutjhout the Reftnim; Procer.

HXf
0,85

0,65 ;

0,6
' 1,1 1,2 1,3 1.4 1,5 1,6 1,7 1.8
Uf
Fig 20 - Colourants throughout the Refiuiny Process (from raw sugar lili fine liquourj
Eight exampies from RAR Reñtier.

•j^;»