J O U R N K t O F I~)aRMI~qTATION A N D B I O E N O I N E E R I N G

V O I . 75, N o . 2, 1 1 5 - 1 2 0 . 1 9 9 3
Spectroscopic Analysis of Polyphenols in White Wines
SHELA GORINSTEIN, I* MOSHE WEISZ, l MARINA ZEMSER, l KIRA TILIS, 2 ALFRED STILLER, 3
ISRAEL FLAM, 3 AND YAKOV GAT a
Department of Pharmaceutical Chemistry, The Hebrew University of Jerusalem, School of Pharmacy, Faculty of
Medicine, Jerusalem 91120, l Food Division, Ministry of Industry and Trade, Jerusalem, 2 and Carmel
Wine Company, Rishon-Le-Zion, 3 Israel
Received 22 May 1992/Accepted 26 November 1992
Spect roscopi c analysis was used t o study the effect o f wi ne processi ng o n phenol i c compos i t i on. Vari ous
classes o f phenol i c c ompomt ds were detected and characterized by ultraviolet ( UV) and Infrared (IR) spectro-
scopy i n whi te grapes o f Sauvi gnon Bl anc and French Col ombard, as wel l as in wi nes prepared f r om these
grapes. Comb/ ned treatment wi t h bent oni t e, egg al bumi n and Pol yci nr AT decreased t he m o u n t s o f
eat ecboi s, f l avoael s , anthoeymains and l euenant hot vani ns. Pol ypbenoi s ( 3 2 - 1 7 ~ ) , ant hoeyanogens ( 64- - 4g~)
and prot ei ns ( 6 2 - 7 7 ~ ) were removed by thi s t echnol ogi cal process. The best results were received when not
onl y wi nes, but al s o must s were pretreated wi th bent oni t e. Compari s ons o f t he pol ypbenoi c ompos i t i ons o f
wi nes made f r om the s ame grape variety grown in different l ocat i ons o f the same vi ntage and bet ween t wo
vintages are reported.
Phenolic compounds contribute in an important man-
ner to the taste, bitterness, and bacteriological effects of
wines. These compounds include catechins, leucoantho-
cyanidins, flavonois, tiavonol glycosides, high-molecular-
weight tannins, hydrocinnamic acid-tartaric esters and
their glucose esters, proanthocyanidins and anthocya-
nidin~, phenofic benzoic and p h e n o l i c c i n n a m i c a c i d s (1, 2).
Turbidity formation caused by the interaction between
must proteins and phenols has been studied by several au-
thors (3-8).
The concentration of phenolic compounds in white
wines depends on the methods of grape processing, grape
crushing and must preparation (9-12).
High levels of polyphenob increase susceptibility to o x i -
d a t i o n , leading to de, creed visual and organoleptic quali-
ties (13). Sauvignon Blanc is the least susceptible to brown-
ing (9, 14). The levels of polyphenols and proteins are
reduced in wine material treated with different adsorbents,
such as bentonite, polyclar, gelatin, egg albumin and
others (8, 15-18). There is a lack of data corresponding to
the quality of final products and the fractions of poly-
phenols and proteins in Israeli white wines. Therefore, two
varieties of white grapes (Sauvignon Blanc and French
Colomhard of 1988 vintage), musts and the correspond-
ing wines were subjected to spectroscopic analysis using
variables selected by correlation to the quality of the final
product.
MATERIALS AND ME T HOD S
T wo t ype s o f Israeli I l r l i ~ Sauvignon Blanc from
three growing areas (Slmalabim, Gshor and Hulda) and
French Colombard (M. Tat). These two varieties of grapes
were selected as the basic grapes for different technological
treatments. All technological processes(crushing, fermen-
tation, racking, and fining) were carried out under indus-
trial conditions for grapes for Sauvignon Blanc at Car-
mel Wine Corporation at Rishon-Le-Zion and for French
Colombard at the same Company at Zichron-Yakov.
* Corresponding author.
Analysis of wines Samples of wines were treated in
the laboratory with either bentonite (=B), egg albumin
(= E) and Polyclar AT (= P)-PVPP-(polyvinylpolypyrroli-
done), or with a mixture of these three additives (B, E
and P) in different variations in order to determine if this
treatment affects the stability of the final product with em-
phasis on the content of polyphenols.
Bentonite was mixed with water at ratios of 1 : 10 and
1: 7 w/v. All details of such fining are summarized in
Tables 1 and 2. The time of fining with different adsorb-
ents was the same as for the industrial conditions (7-
10 d). Juice of French Colomhard grapes was treated with
bentonite (0.5 g//) at 10C during 24 h. Control 1 was the
wine sample before treatment and Control 2 represented
the same wine sample after treatment at the winery (Table
1). Samples of musts and wines were subsampled during
different stages of wine preparation, such as fresh grape
juice, complete juice fermentation, last racking, before
fining, after fining with different adsorbents and also sam-
pies after filtration.
Standard wine and must analysis was carried out using
conventional methods (19). The wine samples were dia-
lyzed against water for 72 h at 4C and then freeze-dried.
Concentrates of wines were diluted to their original
degrees Brix before analysis.
Analysis of phenolic acids
UV spectroscopy Twenty one phenols were chosen
as standards for UV analysis: (eatechin; rutin; quercetin;
fisetin; cinnamic acid; caffeic acid gallic acid; salicylic acid;
p-hydroxybenzoic acid; 2,3-dihydroxybenzoic acid; 2,4-
dihydroxybenzoic acid; 2,5-dihydroxybenzoic acid; 3,4-
dihydrobenzoic acid, 2,4,6-trihydroxybenzoic acid; 2,3,
4-trihydroxybenzoic acid; 4-hydroxyphenylacetic acid;
resorcinol; tannic acid; catechol; caffein and vanillin).
Sample preparation All determinations were carried
out in triplicate. Samples were prepared according to
Tryon at al. (20), with some of our modifications, based on
the solubility of phenols and origin of wine. 5 ml of wine
were diluted to 10Oral with a solution of 12% (v/v)
ethanol and 5~ (v/v) dextrose.
Wine samples which were previously freeze-dried were
1 1 5
116 GORINSTEIN ET AL. J. FI/RI~ITr. BIOENG.,
TABLE 1. ResuRs* of fining with different adsorbents in white wine (Sauvignon Blanc) from grapes collected at Gshor
Indices Control 1 Control 2 Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 Test 7 Test 8
Bentonite (1 : 1 0 ) , g/l
Bentonite (1 : 7), g/ I
Egg albnmin~ mg/I
Polyclar AT, mg/ !
Proteins, ms/ l
Polyphenols, mg/I
Leucoanthocyanin, mg/ i
Cu, mg/i
Fe, mg/l
Stability - 5 oC
+55C
- - +
- -
- -
- - -I-
34.8+1.0 18.2-+0.7 16.5-+0.5 14.8--+0.3 14.5-+0.4 30.5-+1.1 13.7--+0.2 12.5-+0.3 11.2--+0.2 10.2-+0.1
420+9.0 390+7.3 390-+7.1 385-+6.5 383-+7.0 365-+8. 8 370-+6.9 370-+6. 8 360+8.1 350-+7.3
180-+3.1 165-+2. 4 160-+1.9 155-+1.8 153-+2. 0 112-+2.3 150-+1.5 148_+1.7 104-+1. 1 95_+0.9
0.15-+0.09 0.13-+0.08 0.12-+0.10 0.11-+0.10 0.10-+0.09 0.10-+0.09 0.10-+0.09 0.09-+0.01 0.08-+0.03 0.07-+0.01
6.0-+0.2 5.7-+0.1 5.6+0.2 5.2-+0.1 5.1-+0.2 5.0-+0.2 4.9-+0.3 4.7+0.4 4.6-+0.2 4.4+0.2
Unstab. Stab. Stab. Stab. Stab. Stab. Stab. Stab. Stab. Stab.
* Each value is the mean of three determinations -+ SD.
TABLE 2. Results* of finin~ with different adsorbents in white wine (Trench Colombard) from grapes collected at M. Tut
Indices Control I Control 2 Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 Test 7 Test 8
Bentonite (1 : 10), g/ ! - - +
Bentonite (1 : 7), g/ I - -
Egg albumin, rag~! - -
Polyclar AT, mg/ ! - - +
Proteins, rag~! 10.3+0.3 5.5--+0.2 5. 3_+0. 4 5. 1_+0. 2 4.9-+0.1 9.4--+0.4 5.0-+0.3 4.8-+0.2 4.5-+0.4 4.0-+0.3
Polyphenols, mg/l 450-+9.8 380-+8.4 380-+8. 4 375-+8.0 375-+5. 7 330-+6.9 375-+7.9 370-+7.6 320-+6. 8 310-+5.9
Leucoanthocyanin, mg/! 210-+3.6 160-+3.7 150+3.2 148-+2.8 146-+3.1 100-+1. 6 145-+2.9 140-+3. 0 90-+1.8 72-+1.2
Cu, mg/ l 0.15-+0.04 0.12-+0.05 0.10-+0.06 0.08-+0.08 0.08+0.02 0.08-+0.03 0.09-+0.01 0.07_+0.03 0.06+0.01 0.05-+0.01
Fe, mg/i 5.6+0.4 5.3-+0.4 5.1+0.3 5.0-+0.4 4.9-+0.2 4.9+0.1 4.9-+0.5 4.7+0.3 4.2-+0.4 4.0-+0.2
Stability , 5 C Unstab. Stab. Stab. Stab. Stab. Stab. Stab. Stab. Stab. Stab.
+55C
* Each value is the mean of three determinations_+SD.
di ssol ved i n wat er accor di ng t o t he l yophi l i zed wei ght i n
or der t o add 5 mi o f or i gi nal wine, and were t hen di l ut ed
t o 100ml wi t h an et hanol - dext r ose sol ut i on. Phenol s
(quercet i n, r ut i n and f ~ t i n ) as well as phenol i c f r act i ons
o f l yophi l i zed wi ne sampl es were first di ssol ved i n et hanol
and t hen di l ut ed wi t h t he et hanol - dext r ose sol ut i on.
~
t ock sol ut i ons (100 r ag/ / ) o f each phenol were pr epar ed
y di ssol vi ng t he sampl e i n et hanol - dext r ose sol ut i on.
The abs or pt i on o f s t andar d sol ut i ons ( 10mg/ / ) , a s well
as wi nes were meas ur ed wi t h a Wi con 930 Ka ut r on UV
s pect r ophot omet er at a scan r ange o f 250--350 n m and a
scan speed o f 200 u m/ mi n . Al l ul t ravi ol et spect r a were
r ecor ded usi ng et hanol , dext r ose sol ut i on I s a bl ank.
The a mo u n t o f t ot al pol yphenol s ext r act ed wi t h met h-
anol was det er mi ned s pect r ophot omet r i cal l y at 275 nm.
Resor ei nol was used as a s t andar d (21). Phenol i c c om-
pounds were al so det er mi ned by t he Fol i n- Ci ocal t eu
me t hod (22).
I R anal ysi s The I R spect r a o f l yophi l i zed wi ne sam-
ples were meas ur ed by Four i er Tr a ns f or ma t i on I nf r ar ed
Spect r os copy ( FTI R) as a film bet ween t wo KBr pl at es
wi t h a Four i er Tr a ns f or ma t i on (FT) I R Anal ect i nst ru-
ment . The r ecor di ng was done f r o m 4, 000 t o 2, 500 cm -~
wave number . St andar ds f or t ot al pol yphenol cont ent
were pur chased f r o m Si gma Chemi cal Co. (St. Loui s,
USA) .
Pr ot ei n anal ysi s Tot al pr ot ei ns i n must s and wi nes
were det er mi ned i n 1 mi o f t he sampl e whi ch was t r eat ed
wi t h 4 ml Bri l l i ant Bl ue G- 250 f or 10mi n, f ol l owed by
col or i met r y at 595 n m (23, 24).
100 ml o f j ui ce or wi ne at di fferent st ages o f vi ni l i cat i on
were l yophi l i zed and t he wei ght o f dr y subst ance was det er-
mi ned. Dr y pr eci pi t at e was di ssol ved i n acet one 1 : 5
( w/ v) . The a c e t o n ext r act abl e f r act i ons, cont ai ni ng pr o-
teins, were di ssol ved i n var yi ng amount s o f 2- mer capt o-
et hanol (2-ME) f r om 2 - 2 0 ~ f or ma xi mum ext r act i on.
Tot al pr ot ei ns were pr eci pi t at ed al so f r o m 1 1 o f wi ne
by sat ur at i ng t he sol ut i on wi t h a mmo n i u m sul fat e (540
g), l eavi ng it st and over ni ght at 4 C, cent r i f ugi ng at
9, 000 r pm, and t hen di al yzat i on f ol l owed by l yophi l i za-
t i on.
Acet one ext ract abl e and a mmo n i u m sul phat e preci pi -
t abl e pr ot ei ns were i nvest i gat ed i n 10--20~o gr adi ent s b y
s odi um dodecyl sul phat e pol yacr yl ami de gel el ect r opho-
resis ( SDS- PAGE) accor di ng t o Laemml i (25). St andar ds
obt ai ned f r o m Si gma Chemi cal Co. o f 8. 1; 14.2; 20. 0;
24. 0; 29. 0; 36. 0; 45. 0 and 66. 0 kDa were used f or t he mo -
l ecul ar wei ght est i mat i on o f pr ot ei n subuni t s.
Ot her pr ocedur es St abi l i t y t o be a t / c ol d t est i ng was
done at 55C and - - 5 C f or 48 h. The PVPP- t r e a t e d wi nes
were al so subj ect ed t o a f or ced br owni ng test at + 50C
f or 72 h.
Cu and Fe were det er mi ned by at omi c abs or pt i on spec-
t r ophot ome t r y.
RESULTS AND DI SCUSSI ON
Tr eat ment wi t h bent oni t e suspensi on under di fferent
concent r at i ons i n wat er t oget her wi t h t he addi t i on o f egg
al bumi n and Pol ycl ar AT depressed t he a mo u n t o f pol y-
phenol s up t o 1 7 - 3 2 ~ and t he l eucoant hocyani n cont ent
o f t he wines was di mi ni shed t o 48- 64~ c ompa r e d t o initial
cont r ol sampl es (Tabl es 1 and 2).
Our da t a s how si mi l ar amount s o f pol yphenol s and l eu-
coant hocyani ns i n Sauvi gnon Bl anc whi t e wines ma de
f r o m grapes col l ect ed i n Shaal abi m, Gs hor and Hul da .
Ther ef or e ori gi nal r epor t s o f Shaal abi m and Hu l d a have
been omi t t ed i n t hi s r epor t . Differences i n t he phenol i c
make- up o f t he 2 cul t i vars (Tabl es 1 and 2) were small.
The phenol and t anni n cont ent s were i dent i cal wi t h ones
VoL. 75, 1993 UV AND IR SPECTROSCOPY AND WHITE WINES 117
2.0
~
1.0
250 300 350
Wavelength, nm
FIG. 1. UV spectra of some phenolics. 1: Resorcinol; 2:
catechol; 3: catechin; 4: rutin; 5: quercetin; 6: fisetin; 7: cinnamic
acid; 8: caffein; 9: 4-hydroxyphenyl acetic acid; 10: tannic acid; 11:
caffeic acid.
r epor t ed i n t he l i t erat ure (26). However , some aut hor s
have f ound ver y hi gh a mount s o f l eucoant hocyani n con-
t ent in whi t e wines (11, 27).
Mean values o f phenol i c subst ances were l ower i n this
season t han i n t he 1978 vi nt age (6, 15). Fr ench Col omba r d
i nvest i gat ed i n this s t udy was t r eat ed wi t h bent oni t e (Tabl e
2) dur i ng mus t f er ment at i on. The a mount o f t ot al pol y-
phenol s, as well as pr ot ei ns decr eased mor e abr upt l y
(Tabl e 2) t ha n i n sampl es not t r eat ed wi t h adsor bent s
dur i ng mus t f er ment at i on (Tabl e 1). Si mi l ar resul t s have
been r epor t ed b y Ama t i (3) and Yokot s uka et al . (8).
Tot al pol ypbenol s and l eucoant hocyani ns were meas-
ur ed i n all wi ne sampl es t r eat ed wi t h Pol yct ar AT and af t er
agi ng dur i ng f our mont hs . The a mount s o f pol yphenol s
and l eucoant hocyani ns were si mi l ar f or Cont r ol 2 and
Tests 7 and 8 o f wi ne sampl es, pr epar ed f r o m gr apes col -
lected i n f our di fferent l ocat i ons (Tabl es 1 and 2). Br own
col or test was negative i n all sampl es t r eat ed wi t h Pol ycl ar .
The~e dat a ar e si mi l ar t o pr evi ous r epor t s (3, 28). Ther e
is no l i t erat ure da t a avai l abl e on t he phenol i c cont ent o f
Israeli Sauvi gnon Bl anc and Fr ench Co l o mb a r d wi nes
t o whi ch a compar i s on can be made.
TABLE 3. Absorption maxima of some phenols
Phenolic compounds
A b s o r p t i o n p e a k s , n m
1 2 3 4 5 6
Flavonoids
Catechin 328 279
Rutin 299 295 256
Quercetin 326 282 274 253
Fisetin 301 256
Nonflavonoids
Cinnamicacid 340 333 328 270
Catfeicacid 312 287 285
Benzoic acid derivatives
G a l f i c a c i d 3 4 4 3 " 4 0 3 3 3 2 6 1
S a l i c y l i c acid 299 295 256
p-Hydroxybenzoic acid 271
2,3-Dihydroxybenzoic acid 306
2,4-Dihydroxybenzoic acid 342 292
2,5-Dihydroxybenzoic acid 321
3,4-Dihydroxybenzoic acid 341 290 253
2,4,6-Trihydroxybenzoicacid 341 293 255
2,3,4-Trihydroxybenzoicacid 347 295 260
Others
Caffeine 340 333
Catechol 346 335
4-Hydroxyphenylacetic acid 343 325
Resorcinol 335 328
Tannic acid 3 4 8 3 4 0
V a n i l l i n 311
328 324 314 272
328 273
317 306 275
314 309 302 273
338 277
The UV and I R spect r a o f several whi t e Israel i wines
were used t o det er mi ne t hei r phenol i c cont ent , as a resul t
o f P VP P t r eat ment wi t h and wi t hout ot her adsor bent s
(Tabl es 1 and 2). Fi gures 1 and 2 s how t he UV spect r a f or
all phenol s i nvest i gat ed. The charact eri st i c abs or pt i on
da t a o f Figs. 1 and 2 ar e s hown i n Tabl e 3. As can be seen
f r o m Tabl e 3, t he abs or pt i on ma xi ma f or all s t andar ds
were bet ween 350 and 250 nm.
Fi gur e 3 present s t wo wi ne sampl es i n c ompa r i s on wi t h
cat hechi n, caffeic, gallic and t anni c acids, t he mos t
represent at i ve aci ds i n t he phenol i c compos i t i on o f wines.
The wi ne spect r um (Fig. 3) was si mi l ar t o one r epor t ed
by Tr y o n e t al . (1988), wher e t he per cent ages o f phenol s
ads or bed by P VP P were t he f ol l owi ng: quer cet i n (69-73);
r ut i n (14); cat echi n (36); caffeic aci d (70--78); gallic aci d
( 5 7 ) a n d s a l i c y l i c a c i d ( 4 5 ) . T h e a b s o r b a n c e o f w i n e f i n i n g
o ~ 1 . 0
~ o ~ ~
0.5
o
0 250 3oo 350 0.5 . ~ ~ ~ , ~ - ~__. ~_ \ ~ ~
WaveLength. nm 0.0
250 300 350
FIG. 2. UV spectra of benzoic acid derivatives and nonfla-
v o n o i d s . I : R e s o r c i n o l ; 2 : g a l l i c a c i d ; 3 : v a n i l l i n ; 4 : s a l i c y l i c a c i d ; Wavelength, nm
5 : p - h y d r o x y b e n z o i c a c i d ; 6 : 2 , 3 - d i h y d r o x y b e n z o i c a c i d ; 7 : 2 , 4 - F I G . 3 . U V s p e c t r a o f I , T A n n i c a c i d ; 2 , c a l f e i c a c i d ; 3 , g a l l i c
dihydroxybenzoic acid; 8: 2,5-dihydroxybenzoic acid; 9: 3,4-dihydro- acid; 4, catechin; 5, wine treated with bentonite and egg albumin
xybenzoic acid; 10: 2,3,4-trihydroxybenzoic acid; 11: 2,4,6-trihydro- (corresponding to Test 6, Table 1); 6, wine treated with bentonite,
xybenzoic acid. egg albumin and Polyclar (corresponding to Test 8, Table 1).
118 GORI NSTEI N ET AL. J. FI/Kt~N'T. BIOENO.,
with PVPP was lower t han in unt reat ed samples, indicat-
ing the removal of phenols had occurred (Fig. 3).
Tabl e 4 illustrates t hat wine samples revealed absorb-
ance peaks in t he range of all 21 phenolic standards (Figs.
1 and 2, Tabl e 3). The number of peaks in wine samples
was smaller t han in pure standards, suggesting t hat the ex-
tracted compounds were a mixture of phenols.
The phenolic O- H stretch absorbs at 3705-3125cm -1
for phenols, catechols and resorcinols. Aromat i c carbox-
ylic acids as cinnamic, gallic and others O-H stretch ab-
sorbs at 3,335-2,500 cm -1.
IR spectra recorded the following peaks (in cm-1): for
catechin (2643; 2746, 2849; 2937; 3106;); caffeic acid (2541;
3210); gallic acid (2875; 3132; 3544) and tannic acid (3232;
3441).
In wine samples shown in Fig. 4 peaks were recorded
nearly in the same range (cm-1): in samples of wine t reat ed
with bent oni t e and egg albumin (corresponding to Test 6,
Tabl e 1): 2515; 3081; 3389; 3492; 3544 and for samples of
wine t reat ed with bentonite, egg albumin and polyclar (cor-
responding t o Test 8, Tabl e 1): 2566; 2721; 2926; 3184;
3284; 3441; 3698. Ther ef or e t he FTI R spectra of standards
and wine samples were t ot al l y consistent with one anot her
in the O-H-st ret ch region.
Three tested fining agents decreased the amount of poly-
phenols and l eucoant hocyani ns in the order Pol ycl ar
AT > Bentonite > Egg albumin. Leucoant hocyani n cont ent
is an i ndi cat or of wine stability. A wine sample f r om
Shaalabim (Test 8) was the most stable and its amount of
l eucoant hocyani n was the lowest. Most likely proteins
produce t urbi di t y onl y with t anni n fractions. This has
been shown in a model system (8).
All wine samples had <4 0 0 mg / / polyphenols and
showed resistance t o oxi dat i on, findings which correspond
to those of Vacca et al. (13). The decreases in t he amount s
TABLE 4 . Ab s o r p t i o n ma xi ma i n whi t e wi ne ( Sauvi gnon Bl anc)
f r om gr apes col l ect ed at Gs hor
Abs or pt i on peaks, n m
I nvest i gat ed Tr e a t me nt
sampl es 1 2 3
Fr es h j ui ce (L) ~ - - 282
Fr es h j ui ce ( L + D) b - - 278
Ext . phenol s f r om - - 281 279
f r esh j ui ce c (L)
Mus t (L + D) af t er f e r me nt a t i on 278 263
Wi ne ( L + D) be f or e fi ni ng 277 266
Ext . phenol s f r om bef or e fi ni ng 272
wind (L)
Ext . phenol s f r om be f or e fi ni ng 275 273
wi ne (L + D)
Wi ne (L) B (1 : 7)g 282 279
Wi ne ( L + D) B (1 : 7 ) + E h 277 263
Wi ne (ori gi nal ) f B (1 : 7 ) + E + P h 266
Wi ne (L) B (1 : 10)g 281
Wi ne (ori gi nal ) B (1 : 1 0 ) +E 268
Wi ne ( L + D) B (1 : 1 0 ) + E + D 281 276
277
271
a Lyophi l i zed.
b Di al yzed.
c Phenol i c c o mp o u n d s ext r act ed wi t h me t ha nol f r om l yophi l i zed
f r esh j ui ce.
d Phe nol i c c o mp o u n d s ext r act ed wi t h me t ha nol f r om l yophi l i zed
wi ne.
e Phenol i c c ompounds ext r act ed wi t h me t ha nol f r om l yophi l i zed
a nd di al yzed wi ne.
f Fr es h or i gi nal wi ne.
g Fi ni ng wi t h be nt oni t e (1 : 7), g/l.
h Fi ni ng wi t h egg al bumi n, mg/ / ; fi ni ng wi t h Pol ycl ar AT, mg/I.
55 A 55 B
49 ~
33
~. 25
I
ffl
55 ~ m - 4 0 0 1600
W o v e t u r b e r ( c m - ' )
c-
25
I - -
17
9 3 2 b 0 4 ' 0 0 1 6 b 0
WaveLength, (cm -~)
FI G. 4. FTI R spect r a o f A: 1, t anni c aci d; 2, cat echi n; 3,
gallic aci d; B: 4, wi ne t r eat ed wi t h bent oni t e and egg al bumi n ( cor r e-
s pondi ng t o Test 6, Tabl e 1); 5, wi ne t r eat ed wi t h bent oni t e, egg al bu-
mi n and Pol ycl ar ( cor r es pondi ng t o Test 8, Tabl e l ).
(mg//) of Fe and Cu were ar ound 5. 6-3. 0 and 0.15-0.04,
respectively (Tables 1 and 2). These values were lower
t han f or vintage 1978 (6, 15).
The amount of prot ei n decreased in the range of 62 t o
72~ compared t o initial amount s. Accordi ng t o t he results
of isoclectric focusing of proteins in musts of Sauvignon
Blanc and French Col ombard, onl y t wo mai n bands were
f ound at pH 4.0. The wines ferment ed f r om the same
musts showed 2 prot ei n bands at about pH 3.8. The differ-
ences in t he compositions of proteins in wines and musts
were very small. The t ot al number of isoelectric focusing
bands was about 8 between pH 3.8 and 7.0. These results
are similar t o those of ot her aut hor s (29, 30).
SDS-PAOE demonst rat ed t hat t he amount s of must and
wine proteins were about 8; 10; 15; 22; 29 kDa, with mi nor
bands of 45 and 66 kDa. It was observed t hat ferment at i on
as well as the fining process reduced t he t ot al amount and
number of fractions. Using chr omat ogr aphy on Superose
MW
Kt '
1 2 3
FI G. 5. SDS- PAGE o f wi ne pr ot ei ns i n 10-20~0 gr adi ent P AAG.
1, Standard (10 #1); 2, wine (20 #1) treated with bentonite, egg albu-
min and Polyclar (corresponding to Test 8, Table 1); 3, wine (20 ~1)
treated with bentonite, egg albumin and Polyclar (corresponding to
Test 8, wine from grapes collected in Hulda).
VOL. 75, 1993 UV AND IR SPECTROSCOPY AND WHITE WINES 119
12 wi t h di fferent sol vent syst ems, 9 pr ot ei n f r act i ons were
f ound bet ween 190 and I k Da (31). However , Mes r ob e t
al . (32) r epor t ed t hat wi ne pr ot ei ns were separ at ed i nt o 4
f r act i ons, 28; 128; 144; and 160.
Wi ne pr ot ei ns ext r act ed b y a mmo n i u m sul fat e and sepa-
r at ed by SDS- PAGE s howed di ffused and unseparated
bands i n t he r ange bet ween 8 and 20 kDa, while acet one ex-
t r act abl e pr ot ei ns demons t r at ed shar p bands i n t he same
regi on. Acet one ext r act i on was i nvest i gat ed as a me t hod o f
avoi di ng possi bl e i nt erference f r o m pol yphenol i c com-
pounds whi ch have been encount er ed i n t he f r act i onat i on
o f pl ant pr ot ei ns. Two wi ne sampl es (Fig. 5) f r om grapes
col l ect ed i n Gs hor (line 2) and Hul da (line 3) were si mi l ar
f or bands o f 8, 20 and 29 kDa. Test 8 (wine f r om grapes
col l ect ed i n Hul da) showed s ome addi t i onal bands be-
t ween 8 and 14 k Da and ver y weak ones at 45 and 66 kDa.
Slight differences were f o u n d bet ween 24 and 29 kDa. Thi s
sampl e s howed mor e pr ot ei n cont ent bet ween 24 and 29
kDa t han i n t he Gs hor sampl e (line 2, Fi g. 5). These dat a
cor r es pond exact l y t o t he a mount o f pr ot ei n (test 8 i n
Tabl e 1 and i n wi ne f r om gr apes col l ect ed i n Hul da) , as
well as t o t he st abi l i t y o f wi ne sampl es. Some separ at ed
sampl es ar e not s hown i n t hi s paper because t he bands
were si mi l ar t o ones whi ch have been present ed. The or der
o f adsor bent s used i n t hi s s t udy f or pr ot ei n t r eat ment is
t he f ol l owi ng; Pol ycl ar ~ Bent oni t e ~ Egg Al bumi n.
These studies ma ke it possi bl e t o est abl i sh opt i mi zi ng
profiles o f Israeli whi t e wines wi t h regards t o phenol i c sub-
st ances and pr ot ei ns whi ch can t hen be used as i ndi cat or s
o f wi ne qual i t y.
Pr ot ei n haze seems t o be caused by t he r emoval o f l ow
mol ecul ar wei ght pr ot ei ns by bent oni t e t r eat ment . These
resul t s are c ompa r a bl e wi t h t hose o f di fferent aut hor s f or
var i ous t ypes o f wi ne whi ch f ound t hat t he values obt ai ned
are wi t hi n a r easonabl e r ange (8, 17, 18, 29, 33).
Di fferences i n t he t hr ee Sauvi gnon Bl anc gr ape sampl es
were smal l and coul d general l y be at t r i but ed t o cl i mat i c
vi t i cul t ural condi t i ons (different t i mes and areas o f grapes
col l ect i on) and some slight differences dur i ng fining. Com-
posi t i on was si mi l ar wi t hi n an i ndi vi dual gr ape vari et y
(different areas o f col l ect i on) and differed bet ween vari e-
ties ( Sauvi gnon Bl anc and Fr ench Col ombar d) . Dur i ng t he
t en year s bet ween vi nt ages 1978 and 1988 t her e has been an
i mpr ovement i n i rri gat i on, as well as var i at i ons i n cl i mat e.
However , t hese f act or s had ver y little i nfl uence on t he qual -
i t y o f t he final pr oduct . Tr eat ment wi t h di fferent adsor b-
ent s i nfl uenced t he qual i t y and st abi l i t y o f whi t e wines.
UV and I R s pect r os copy can be appl i ed t o grapes and
wines wi t h sat i sf act or y results. Fi ni ng wi t h different adsor b-
ent s was effective at decreasi ng t he a mount o f phenol i cs.
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