J. Agric. Food Chem.

1999, 47, 21092115

2109

P h e n o lic Au to x id a tio n Is Re s p o n s ible fo r Co lo r D e g ra d a tio n in

P ro c e s s e d Ca rro t P u re e
S. T. Ta lcot t a n d L. R. H owa r d*,
Depa r t m en t of H or t icu lt u r a l Scien ces, Texa s A&M Un iver sit y, College St a t ion , Texa s 77843-2133

St r a in ed ca r r ot s wer e t h er m a lly pr ocessed wit h r edu ced oxygen pr et r ea t m en t s a n d exposed t o

eleva t ed st or a ge t em per a t u r es t o a cceler a t e ph ysicoch em ica l ch a n ges (40 C for 4 weeks). St r a in ed
ca r r ot pr et r ea t m en t s pr ior t o pr ocessin g in clu ded a n it r ogen spa r ge (N 2 ), bla n ch /fr ozen wit h n it r ogen
spa r ge (BF N 2 ), oxygen spa r ge (O 2 ), a n d a con t r ol (C) t h a t r eceived n o pr et r ea t m en t . Ch a n ges in
color , t ot a l solu ble ph en olics, t ot a l ca r ot en oids, ph en olic a cid m olecu la r weigh t , su ga r s, a n d pH wer e
m on it or ed du r in g st or a ge. Gr ea t er losses of color , t ot a l solu ble ph en olics, a n d t ot a l ca r ot en oids
occu r r ed in con trol a n d O 2-spa rged sa mples a s com pa r ed t o N 2-spa r ged a nd BF N 2 sa m ples. Molecu la r
weigh t of ph en olic a cids wa s lower in n it r ogen -spa r ged sa m ples t h a n con t r ol a n d oxygen -spa r ged
sa m ples. P h en olic polym er iza t ion du e t o a u t oxida t ion wa s r espon sible for color loss in pr ocessed
st r a in ed ca r r ot s. P r ocessin g t r ea t m en t s t h a t r edu ce r esidu a l oxygen m a y r esu lt in bet t er color
r et en t ion a ft er pr ocessin g a n d du r in g st or a ge. Det er m in in g t h e m ech a n ism (s) a n d m a gn it u de of
t h ese r ea ct ion s a r e im por t a n t for devisin g st r a t egies t o pr even t qu a lit y loss in st r a in ed ca r r ot s.
Ke y w o rd s : S train ed carrots; ph en olic acid s; au toxid ation ; color; caroten oid s
INTRODUCTION

Br own color for m a t ion in pr ocessed st r a in ed ca r r ot s

is per ceived a s a qu a lit y defect , a n d m et h ods t o pr eser ve
fr esh ca r r ot color a r e im por t a n t for im pr ovin g con su m er
a ccept a n ce. Ma cer a t ion of ca r r ot r oot s followed by
h ea t in g in h oldin g t a n ks ca u ses oxida t ive ch a n ges t h a t
r esu lt in br own color a n d possible off-fla vor for m a t ion .
P r ocessin g in en closed syst em s r esu lt ed in bet t er fla vor
scor es for pea r s t h a t wer e a t t r ibu t ed t o fa st in a ct iva t ion
of en zym es a n d essen ce ca pt u r in g (Leon a r d et a l., 1976).
Nu m er ou s br own in g r ea ct ion s in food syst em s h a ve
been attributed to changes in pH (Cilliers and Singleton,
1990; Yeo a n d Sh iba m ot o, 1991) a n d t im e a n d t em per a t u r e of pr ocessin g (F r iedm a n a n d Moln a r -P er l, 1990;
Reyes et a l., 1982). Alka li t r ea t m en t of cooked, gr a t ed
ca r r ot s r esu lt ed in pr odu ct da r ken in g a s sh own by
decr ea sed ligh t n ess va lu es (Ar ch a n a et a l., 1994). Rea ct ion r a t es for Ma illa r d a n d ph en olic n on en zym a t ic
br own in g (NE B) a r e gr ea t ly in flu en ced by t em per a t u r e
a n d pH . Th er m a l pr ocessin g of low a cid foods, in volvin g
eleva t ed t em per a t u r es for ext en ded per iods of t im e,
im pa ct s t h e over a ll qu a lit y of ca r r ot s (Sim on , 1985).
Ma illa r d br own in g r ea ct ion s in st r a in ed ca r r ot s h a ve
been pr oposed bu t n ot con clu sively pr oven . Ma illa r d
br own in g wa s post u la t ed t o con t r ibu t e t o color , n u t r itional, and flavor losses in retorted strained carrots (Luh
et a l., 1969), a n d st u dies r epor t in g loss of r edu cin g
su ga r s a n d a m in o a cids in pr ocessed ca r r ot s wer e
con sist en t wit h t h is t ype of br own in g (H owa r d et a l.,
1996; Tor ibio a n d Loza n o, 1986). H owever , r epor t ed
losses of a m in o a cids in m a n y food syst em s ca n r esu lt
* To wh om cor r espon den ce sh ou ld be a ddr essed
[ph on e, (501)575-2978; fa x, (501)575-2165; e-m a il, lu keh @
com p.u a r k.edu ].
P r esen t a ddr ess: Un iver sit y of Ar ka n sa s, In st it u t e of F ood
Scien ce a n d E n gin eer in g, 272 You n g Ave, F a yet t eville, AR
72704.

fr om com plexa t ion wit h o-qu in on es, wh ich a r e in t er m edia t es in ph en olic a cid oxida t ion (Ch eft el et a l., 1985).
Sin ce Ma illa r d r ea ct ion s do n ot occu r ext en sively below
pH 6.0 (r et a il ca r r ot pu r ee pH 5.0-5.5) or a t com m er cia l
st or a ge t em per a t u r es, ot h er sou r ces of br own in g m a y
be sign ifica n t .
P h en olic polym er iza t ion r ea ct ion s du e t o a u t oxida t ion
ca n r esu lt in br own -color ed pigm en t s t h a t a r e det r im en t a l t o pr ocessed food qu a lit y. Br own in g r a t es of
ca ffeic a cid solu t ion s a r e pH depen den t wit h yellow
pigm en t s for m ed in it ia lly, followed by br own pigm en t s
a s pH in cr ea ses (Cillier s a n d Sin glet on , 1990). Th e
lim it in g fa ct or for ph en olic a u t oxida t ion is t h e pr esen ce
a n d con cen t r a t ion of t h e ph en ola t e ion , wh ich declin es
wit h decr ea sin g pH (Cillier s a n d Sin glet on , 1989). Th e
ph en ola t e ion is a h igh ly r ea ct ive species t h a t ca t a lyzes
oxida t ive r ea ct ion s in ph en olic a cid syst em s. Acidified
ph en olic solu t ion s will gen er a lly r em a in color less u n less
polym er iza t ion occu r s dir ect ly fr om m olecu la r oxygen
pr esen t in t h e syst em . Th er efor e, oxygen exclu sion
cou pled wit h pr odu ct pH is im por t a n t for con t r ollin g
ph en olic polym er iza t ion r ea ct ion s (Bu ch eli a n d Robin son , 1994). Ma jor ph en ols in ca r r ot s in clu de ch lor ogen ic,
ca ffeic, a n d p-h ydr oxyben zoic a cids a lon g wit h n u m er ou s cin n a m ic a cid der iva t ives (Ba bic et a l., 1993). Act in g
a s a ph yt oa lexin , ph en olic a cids m a y in cr ea se u p t o
7-fold in ca r r ot peel du e t o a biot ic st r ess du r in g post h a r vest h a n dlin g a n d st or a ge (Sa r ker a n d P h a n , 1979).
E xclu din g oxygen in low a n d m ediu m a cid food syst em s
m a y pr even t con den sa t ion of ph en olic a cids, r esu lt in g
in gr ea t er color r et en t ion . Th is st u dy wa s u n der t a ken
t o explor e t h e con t r ibu t ion of ph en olic NE B r ea ct ion s
t o color degr a da t ion of pr ocessed st r a in ed ca r r ot s.
MATE RIALS AND ME TH ODS
Ma te ria ls a n d P ro c e s s in g . Ca r r ot s (Dau cu s carota L.)
wer e pu r ch a sed fr om a loca l m a r ket a n d st or ed a t 4 C u n t il
pr ocessed. Root s wer e wa sh ed, h a n d-peeled, cu t in t o 2-cm

10.1021/jf981134n CCC: $18.00 1999 American Chemical Society

Published on Web 04/28/1999

2110 J. Agric. Food Chem., Vol. 47, No. 5, 1999

pieces, st ea m bla n ch ed for 30 m in , a n d cooled in a n ice wa t er
ba th . Roots were then combined with 75% wa ter (ca rrot weight
a ft er bla n ch in g) a n d blen ded in t o a pu r ee u sin g a kit ch en sca le blen der . All blen ds wer e com bin ed t o cr ea t e a h om ogen eou s pooled sa m ple. Tr ea t m en t s a pplied t o su bsa m ples
in clu ded (1) a va cu u m /n it r ogen spa r ge (N 2 ) wh er e t h e pu r ee
wa s dea er a t ed for 10 m in in a 4-L sidea r m fla sk u n der a
va cu u m pr essu r e of 635 m m H g a n d spa r ged for 20 m in wit h
n it r ogen ga s, a n d t h e ja r h ea dspa ce wa s flu sh ed wit h n it r ogen
pr ior t o ca ppin g; (2) a n oxygen spa r ge (O 2 ) wh er e t h e sa m ple
wa s spa r ged for 20 m in , a n d t h e ja r h ea dspa ce wa s flu sh ed
wit h oxygen pr ior t o ca ppin g; a n d (3) a bla n ch /fr ozen wit h
n it r ogen spa r ge (BF N 2 ) t r ea t m en t wh er e peeled wh ole ca r r ot s
wer e st ea m bla n ch ed, cooled in ice wa t er , pa cka ged in polyet h ylen e pou ch es, a n d fr ozen a t -20 C. Aft er bein g t h a wed
t o r oom t em per a t u r e, ca r r ot s wer e blen ded t o a pu r ee a n d
t r ea t ed in a m a n n er sim ila r t o t h e N 2 -spa r ged sa m ples. A
con t r ol (C) wa s pr ocessed dir ect ly fr om t h e pooled pu r ee wit h
n o pr et r ea t m en t . F or a ll t r ea t m en t s, ca r r ot pu r ee wa s h a n dfilled in t o 11-oz gla ss ja r s, h ea t ed t o 60 C, a n d t h er m a lly
pr ocessed a t 121 C for 30 m in in a r ot a r y r et or t (St ock P ilot Rot or 900, St ock Am er ica In c., Milwa u kee, WI) a t 19 r pm (F o
> 6). An u n pr ocessed pu r ee sa m ple wa s a n a lyzed t o det er m in e
ph ysicoch em ica l ch a n ges du r in g pr ocessin g. Tr ea t ed sa m ples
wer e st or ed 4 weeks a t 40 C a n d a n a lyzed 0, 1, 2, 3, a n d 4
weeks a ft er pr ocessin g for ph ysicoch em ica l a t t r ibu t es.
In a sepa r a t e exper im en t , ca r r ot pu r ee wa s t h er m a lly
pr ocessed, a s descr ibed a bove, t o sim u la t e a n a t u r a lly h igh
occu r r en ce of ph en olic a cids or ca r r ot s t h a t h a d exper ien ced
st r ess-in du ced syn t h esis of ph en olic a cids. Two t r ea t m en t s, a
con t r ol a n d a ph en olic spike, wer e a pplied t o m on it or t h e r a t e
of ph en olic oxida t ion a n d su bsequ en t ph ysicoch em ica l ch a n ges
du r in g st or a ge. P h en olics spiked in t o r a w ca r r ot pu r ee in clu ded equ a l a m ou n t s of gen t isic, p-h ydr oxyben zoic, m -cou m a r ic, o-cou m a r ic, a n d ca ffeic a cids (1250 m g/kg t ot a l).
Ca r r ot s wer e t h er m a lly pr ocessed a n d m on it or ed weekly
a ga in st a n on spiked con t r ol for 4 weeks a t 40 C.
To ch em ica lly sim u la t e st r a in ed ca r r ot pu r ee, a m odel
solu t ion con t a in in g 3 g/100 g su cr ose, 0.5 g/100 g glu cose, 0.5
g/100 g fr u ct ose, 50 m g/L ca ffeic a cid, 50 m g/L syr in gic a cid,
100 m g/L cyst ein e a n d cit r ic/m a lic a cids (1:1 m ixt u r e for pH
a dju st m en t ) wa s h ea t ed for 2 h a t 100 C t o sim u la t e r esiden ce
time of ca rrots in batch and hold tanks in a commercia l setting.
Va r ia t ion s in m odel solu t ion pH r a n ged fr om 5 t o 7.0. P h en olic
br own in g wa s differ en t ia t ed fr om Ma illa r d br own in g by t h e
exclu sion of cyst ein e in iden t ica l m odels a n d ca lcu la t in g t h e
differ en ce in a bsor ba n ce a t 420 n m .
P h y s ic o c h e m ic a l An a ly s is . Color , t ot a l solu ble ph en olics,
t ot a l ca r ot en oids, m olecu la r weigh t of ph en olic a cids, su ga r s,
a n d pH wer e m ea su r ed in pr ocessed st r a in ed ca r r ot s. Ca r r ot
pH was measured using a Corning model 125 pH meter. Brown
color for m a t ion in m odel syst em s wa s m ea su r ed a t 420 n m
(Ga r cia et a l., 1992). Tr ist im u lu s color wa s m ea su r ed u sin g a
H u n t er La bsca n m odel 5100 color im et er . H u n t er a* a n d b*
va lu es wer e con ver t ed t o h u e (a r ct a n b*/a*) a n d ch r om a (a*2
+ b*2 )0.5 va lu es. Tot a l wa t er -solu ble ph en olics wer e ext r a ct ed
in deion ized wa t er (5 g/50 m L) a n d m ea su r ed u sin g t h e F olin Cioca lt ea u a ssa y a s descr ibed by Swa in a n d H illis (1959).
Su ga r s wer e ext r a ct ed in wa t er (5 g/50 m L) u sin g a Tekm a r
m odel TP 18 t issu em izer (Cin cin n a t i, OH ). Aft er filt er in g
t h r ou gh Mir a clot h (Ca lbioch em , Sa n Diego, CA), t h e isola t e
wa s pa ssed t h r ou gh a pr ep-colu m n con t a in in g 1 g of 200-400
mesh Bio-Rex 5 anion-exchange resin (Bio-Rad, Richmond, CA)
t o r em ove or ga n ic a cids. Sa m ples wer e filt er ed t h r ou gh a 0.45m filter and injected into a Spectra Physics model P100 HPLC
equ ipped wit h a 300 7.8 m m Am in ex H P X-87C (Bio-Ra d,
Rich m on d, CA) ca r boh ydr a t e colu m n h ea t ed a t 85 C. Mobile
ph a se con sist in g of 100% Milli-Q wa t er , r u n isocr a t ica lly a t
0.8 m L/m in , wa s m on it or ed wit h a Sh odex m odel RI-71
r efr a ct ive in dex det ect or . Su cr ose, glu cose, a n d fr u ct ose wer e
qu a n t ified u sin g ext er n a l st a n da r ds.
Tot a l ca r ot en oids wer e ext r a ct ed (2 g/25 m L) wit h a solu t ion
con t a in in g a cet on e/et h a n ol (1:1) wit h 200 m g/L BH T a dded.
Samples were extracted in the dark, filtered through Whatman
No. 4 filt er pa per , a n d wa sh ed u n t il t h e r esidu e wa s color less.

Talcott and Howard

Sa m ples wer e a dju st ed t o 100 m L, a n d t h e a bsor ba n ce wa s
m ea su r ed u sin g a H ewlet t -P a cka r d 8452A diode a r r a y spect r oph ot om et er . Tot a l ca r ot en oids wer e ca lcu la t ed a ccor din g t o
Gr oss (1991) u sin g t h e equ a t ion (AV 10 6 )/(A 1% 100G),
wh er e A is t h e a bsor ba n ce a t 470 n m , V is t h e t ot a l volu m e of
ext r a ct , A 1% is t h e ext in ct ion coefficien t for a m ixt u r e of
solven t s a r bit r a r ily set a t 2500, a n d G is t h e sa m ple weigh t
in gr a m s.
Molecu la r weigh t det er m in a t ion of solu ble ph en olic com pou n ds wa s per for m ed a t 4 C u sin g Seph a r ose CL-6B a ga r ose
r esin (Sigm a Ch em ica l Co., St . Lou is, MO) pa cked in a gla ss
colu m n (4.5 cm 100 cm ). Solu ble ph en olic a cid isola t es wer e
obt a in ed by cen t r ifu gin g st r a in ed ca r r ot s u n t il su per n a t a n t
cou ld be pa ssed t h r ou gh a 0.45-m filt er . Deion ized wa t er
ca r r ied 3 m L of st r a in ed ca r r ot isola t e t h r ou gh t h e colu m n ,
a n d fr a ct ion s wer e collect ed ever y 125 dr ops u sin g a dr op
cou n t er . Dext r a n st a n da r ds wer e u sed t o det er m in e m olecu la r
weigh t dist r ibu t ion s fr om t h e colu m n a n d fr a ct ion s a n a lyzed
for t ot a l ca r boh ydr a t e (Du Bois et a l., 1956) by m ixin g 1 m L
fr om ea ch fr a ct ion wit h 1 m L of 5% ph en ol a n d 5 m L of
con cen t r a t ed H 2 SO 4 in a t est t u be. Tu bes wer e h ea t ed for 10
m in a t 80 C a n d cooled, a n d t h e a bsor ba n ce wa s r ea d a t 490
n m . F r a ct ion s fr om t h e ca r r ot isola t e wer e m ea su r ed a t 280
n m (Lit r idou et a l., 1996), a n d t h e da t a wer e r epor t ed a s t ot a l
solu ble ph en olics.
S ta tis tic a l An a ly s is . Th r ee ja r s fr om t h e fou r t r ea t m en t s
were a na lyzed in triplica te a t ea ch stora ge time. Chemical data
wer e a n a lyzed by a n a lysis of va r ia n ce (SAS In st it u t e, In c.,
1985), a n d m ea n sepa r a t ion wa s con du ct ed u sin g Du n ca n s
m u lt iple r a n ge t est (P < 0.05). St epwise lin ea r r egr ession
a n a lysis wa s per for m ed t o pr edict ca r r ot color fr om ch em ica l
da t a .
RE SULTS AND DISCUSSION

Ch e m ical An alysis. Browning reactions in processed

st r a in ed ca r r ot s wer e pr im a r ily r ela t ed t o ph en olic a cid
con den sa t ion wit h a m in or con t r ibu t ion fr om ca r ot en oid
oxida t ion . Ch a n ges in color , pH , t ot a l solu ble ph en olics,
ca r ot en oids, a n d su cr ose wer e sign ifica n t ly a ffect ed by
st or a ge t im e a n d t r ea t m en t (P < 0.05). A declin e in
solu ble ph en olic con t en t wa s con sist en t wit h a n in cr ea sed level of oxygen in cor por a t ed in t o t h e sa m ples
(F igu r e 1A). Sin ce n o en zym e a ct ivit y r em a in s in
t h er m a lly pr ocessed pr odu ct s, oxida t ion of ph en olic
a cids occu r r ed via a u t oxida t ion r ea ct ion s. Levels of
wa t er -solu ble ph en olics declin ed t h r ou gh ou t t h e st u dy
wit h t h e gr ea t est losses occu r r in g du r in g t h er m a l
pr ocessin g. Th er m a l pr ocessin g a ccou n t ed for 58.5% of
t ot a l ph en olic a cids lost in t h e st u dy for t h e con t r ol,
27.2% in N 2 -spa r ged, 27.8% in BF N 2 , a n d 51.4% in O 2 spa r ged sa m ples. Losses in dica t e t h a t u n it oper a t ion s
t h a t exclu de oxygen pr ior t o t h er m a l pr ocessin g a r e
cr it ica l for m a in t a in in g st r a in ed ca r r ot color . P h en olic
a cids, a s m on it or ed by t h e F olin -Cioca lt eu a ssa y, wer e
polym er ized en ou gh t o becom e wa t er -in solu ble sin ce t h e
a ssa y will det ect a n y solu ble r edu cin g com pou n ds
including some conjugated phenols and reducing suga rs.
Th e declin e in pH wa s gr ea t est a ft er t h er m a l pr ocessin g
(0.9 u n it ), wh ich ca n be a t t r ibu t ed t o a r edu ct ion in
pa r t icle size a n d su bsequ en t r elea se of or ga n ic a cids or
a ddit ion a l u n kn own r ea ct ion s (F igu r e 1B). Th e pH
con t in u ed t o declin e du r in g st or a ge a t eleva t ed t em per a t u r e a n d r ea ch ed it s lowest va lu e of 5.27 in O 2 -spa r ged
sa m ples. Decr ea sed pH is det r im en t a l t o over a ll ca r r ot
fla vor , a n d a ddit ion a l wor k is n eeded t o u n der st a n d t h e
m ech a n ism r espon sible for t h e declin e in t h er m a lly
pr ocessed pr odu ct s.
Gr ea t er ca r ot en oid r et en t ion wa s obser ved in N 2 spa r ged a n d BF N 2 t r ea t m en t s a s com pa r ed t o O 2 spa r ged a n d con t r ol sa m ples (F igu r e 1C). Ca r ot en oid

Effect of Phenolic Acids on Carrot Puree Color

J. Agric. Food Chem., Vol. 47, No. 5, 1999 2111

F ig u re 1. E ffect s of pr et r ea t m en t on (A) t ot a l solu ble ph en olics, (B) pH , a n d (C) t ot a l ca r ot en oids in st r a in ed ca r r ot s. Tr ea t m en t s

in clu de n o pr et r ea t m en t (con t r ol), n it r ogen spa r ge (N 2 spa r ge), bla n ch ed/fr ozen wit h n it r ogen spa r ge (BF N 2 ), a n d oxygen spa r ge
(O 2 spa r ge). Un pr ocessed pu r ee dem on st r a t es ch a n ges du r in g t h er m a l pr ocessin g. Ba r s r epr esen t st a n da r d er r or of t h e m ea n .

con cen t r a t ion s declin ed expect edly in con t r ol (6.4%) a n d

O 2 -spa r ged (40.6%) a s com pa r ed t o N 2 t r ea t m en t s.
Gr ea t er r et en t ion in N 2 -t r ea t ed sa m ples dem on st r a t ed
t h a t O 2 exclu sion fr om st r a in ed ca r r ot s wa s ben eficia l
for ca r ot en oid a n d color r et en t ion . H u e a n gle va lu es
(F igu r e 2A) sign ifica n t ly in cr ea sed a ft er t h er m a l pr ocessing in O 2-sparged samples and then declined throughou t t h e st u dy, wh ich m a y be du e t o t h e for m a t ion of
color ed pigm en t s fr om NE B ph en olic polym er iza t ion
r ea ct ion s. Nit r ogen spa r gin g t r ea t m en t s r esu lt ed in
lower h u e a n gles a ft er t h er m a l pr ocessin g a n d declin ed
at a slightly slower rate over time as compared to control
a n d O 2 -spa r ged sa m ples. An eleva t ed h u e a n gle in O 2 spa r ged a n d con t r ol sa m ples wa s in dica t ive of in it ia l
oxida t ion pr odu ct s (yellow pigm en t s) of ph en olic a cids
t h a t wer e pr even t ed fr om polym er izin g in N 2 -t r ea t ed
sa m ples. Th er efor e, a h igh h u e a n gle t h a t declin ed over
t im e wa s in dica t ive of pr odu ct da r ken in g a s m on om er
u n it s polym er ized in t o br own pigm en t s.
Declin es in pH , solu ble ph en olics, a n d t ot a l ca r ot en oids a ll cor r ela t ed wit h decr ea sed ch r om a va lu es (r 2
) 0.49, 0.90, a n d 0.70, r espect ively) in st r a in ed ca r r ot s
(F igu r e 2B). Non pola r a n d polym er ized ph en olic a cids
m a y h a ve a ffin it y for t h e lipid fr a ct ion in ca r r ot s, wh ich
con t a in ca r ot en oids, dir ect ly lower in g ch r om a va lu es or
u pon t h eir su bsequ en t oxida t ion . Th is a ct ion wou ld
ser ve t o dull t he vivid ora n ge color fou n d in n on -oxidized

pu r ees. Lin ea r r egr ession a n a lysis dem on st r a t ed t h a t

h u e a n gle a n d ch r om a wer e sign ifica n t ly a ffect ed by t h e
levels of solu ble ph en olics a n d pu r ee pH (R 2 ) 0.91 a n d
0.83, r espect ively), wh ich dem on st r a t es t h e im por t a n ce
of st r ict oxygen exclu sion pr ior t o t h er m a l pr ocessin g.
Th e in a bilit y t o com plet ely r ever se color for m ed a ft er
oxygen spa r gin g of ph en olic a cid m odel syst em s h a s
been dem on st r a t ed in ou r la b; h owever , visible color
for m ed by a lka li a ddit ion ca n be r ever sed if t h e pu r ee
is a cidified wit h in a sh or t per iod of t im e. Sim ila r color ed
pigm en t s for m ed in a ca ffeic a cid m odel syst em a bove
pH 5.60 a n d wer e a t t r ibu t ed t o qu in on e for m a t ion
du r in g oxida t ion (F u lcr a n d a t a l., 1994). St r a in ed ca r r ot s t h a t wer e h eld in lon g-t er m st or a ge (>2 yr ) exh ibit ed im pr oved color (L * ) +1.59) wh en a cidified, in dicating that irreversible pigments from Maillard browning
wer e n ot t h e sole sou r ce of pigm en t a t ion for m ed. A
sim ila r in cr ea se in ligh t n ess du e t o a cidifica t ion wa s
dem on st r a t ed by Ar ch a n a et a l. (1994) t h a t wa s a t t r ibu t ed t o ch a n ges in ca r ot en oid isom er s pr esen t .
Ch a n ge in pigm en t a t ion wa s a lso n ot iced in pH -a lt er ed
sweet pot a t o pu r ee wh er e n or m a l color r et u r n ed a s t h e
sa m ple pH wa s a dju st ed ba ck t o it s or igin a l va lu e (Ice
et a l., 1980). Ligh t n ess (L *) va lu es in st r a in ed ca r r ot s
declin ed con t in u a lly t h r ou gh ou t t h e sh elf life st u dy wit h
t h e la r gest declin e occu r r in g du r in g pr ocessin g (F igu r e
2C). H igh L * va lu es in dica t e a ligh t er color wh ile

2112 J. Agric. Food Chem., Vol. 47, No. 5, 1999

Talcott and Howard

F ig u re 2. E ffect s of pr et r ea t m en t on (A) h u e a n gle, (B) ch r om a , a n d (C) H u n t er L * of st r a in ed ca r r ot s. Tr ea t m en t s in clu de n o

pr et r ea t m en t (con t r ol), n it r ogen spa r ge (N 2 spa r ge), bla n ch ed/fr ozen wit h n it r ogen spa r ge (BF N 2 ), a n d oxygen spa r ge (O 2 spa r ge).
Un pr ocessed pu r ee dem on st r a t es ch a n ges du r in g t h er m a l pr ocessin g. Ba r s r epr esen t st a n da r d er r or of t h e m ea n .

decr ea sed ch r om a a n d h u e va lu es cor r espon d t o da r ker

color s (Ya n g a n d Ya n g, 1987). Ba sed on ligh t n ess va lu es
of u n pr ocessed st r a in ed ca r r ot s, con t r ol sa m ples declin ed 9.8% a ft er pr ocessin g a s com pa r ed t o 5.8% a n d
5.3% losses for N 2 a n d BF N 2 sa m ples, r espect ively.
Du r in g st or a ge, O 2 -spa r ged sa m ples slowly blea ch ed
a n d wer e a lm ost com plet ely wh it e fr om t h e h ea dspa ce
t o t h e cen t er of t h e pr odu ct , r esu lt in g in eleva t ed
ligh t n ess va lu es. Th e pr esen ce of oxygen likely in it ia t ed
ext en sive ph en olic a n d lipid oxida t ion , r esu lt in g in t h e
for m a t ion of r ea ct ive per oxides r espon sible for ca r otenoid losses (R 2 ) 0.57). However, the regression model
for ligh t n ess wa s n ot gr ea t ly im pr oved wh en t h e O 2
t r ea t m en t wa s r em oved fr om t h e m odel (R 2 ) 0.64).
Com pa r ed t o con t r ol a n d O 2 -spa r ged sa m ples, N 2 a n d
BF N 2 t r ea t m en t s r esu lt ed in h igh er ligh t n ess va lu es,
dem on st r a t in g a r ela t ion sh ip wit h t h e degr ee of oxida t ion .
Molecu la r weigh t (MW) det er m in a t ion of wa t er solu ble ph en olics wa s per for m ed on sa m ples isola t ed
fr om da y 0 (im m edia t ely a ft er t h er m a l pr ocessin g) a n d
a ft er 4 weeks st or a ge (F igu r es 3 a n d 4, r espect ively).
Molecu la r weigh t fr a ct ion s n ea r t h e m a xim u m a bsor b-

a n ce a t 280 n m wer e qu a n t ified by t h e F olin -Cioca lt eu

a ssa y a n d cor r espon ded t o t h e r et en t ion of low m olecu la r weigh t ph en olic com pou n ds (Ta ble 1). Nit r ogen a n d
BF N 2 t r ea t m en t s h a d h igh er a m ou n t s of low m olecu la r
weigh t ph en olics (252 a n d 253 m g/kg, r espect ively) t h a n
con t r ol a n d O 2 -spa r ged sa m ples (223 a n d 207 m g/kg,
r espect ively) a ft er pr ocessin g, in dica t in g t h a t ph en olic
polym er iza t ion wa s r edu ced wit h oxygen exclu sion .
Molecular weight determination of phenolic acids served
t o fu r t h er iden t ify t h eir r ole in pr odu ct da r ken in g a n d
is a ppa r en t ly r ela t ed t o ph en olic polym er iza t ion r ea ct ion s r esu lt in g in t h e loss of t h ese com pou n ds.
Su cr ose con cen t r a t ion s wer e sligh t ly h igh er in BF N 2
sa m ples t h a n con t r ol, O 2 -spa r ged, a n d N 2 -spa r ged
sa m ples (da t a n ot sh own ). Th is r esu lt m a y be a t t r ibu t ed
t o t h e fr eeze-t h a w cycle t h a t r esu lt ed in cellu la r
disr u pt ion a n d gr ea t er liber a t ion of su ga r s. Ya n (1989)
a lso fou n d t h a t pr ocessin g fr ozen ca r r ot s in t o pu r ee
r esu lt ed in h igh er Br ix levels t h a n pu r ee obt a in ed fr om
n on fr ozen ca r r ot s. Glu cose a n d fr u ct ose con cen t r a t ion s
wer e u n a ffect ed by pr ocessin g t r ea t m en t s, in dica t in g
t h a t r edu cin g su ga r s wer e n ot con su m ed in Ma illa r d
r ea ct ion s. In con t r a st , H owa r d et a l. (1996) r epor t ed a

Effect of Phenolic Acids on Carrot Puree Color

F ig u re 3. E ffect of pr et r ea t m en t on m olecu la r weigh t dist r ibu t ion (kDa ) of ph en olic a cids in st r a in ed ca r r ot s im m edia t ely
a ft er t h er m a l pr ocessin g. Tr ea t m en t s in clu de n o pr et r ea t m en t
(con t r ol), n it r ogen spa r ge (N 2 spa r ge), bla n ch ed/fr ozen wit h
n it r ogen spa r ge (BF N 2 ), a n d oxygen spa r ge (O 2 spa r ge).

loss of r edu cin g su ga r s in ca r r ot pu r ee, wh ich wer e

h ypot h esized t o be con su m ed in Ma illa r d br own in g
r ea ct ion s. Or ga n ic a cids r em a in ed u n ch a n ged t h r ou gh ou t t h e st u dy a n d wer e u n a ffect ed by pr ocessin g t r ea t m en t s (da t a n ot sh own ).
St r a in ed ca r r ot sa m ples spiked wit h ph en olic a cids
followed sim ila r t r en ds for color degr a da t ion a s descr ibed in t h e a cceler a t ed sh elf life st u dy (da t a n ot
sh own ). Solu ble ph en olic ch a n ges in con t r ol a n d spiked
sa m ples declin ed a t a con st a n t r a t e wit h sign ifica n t
losses obser ved a ft er pr ocessin g (9.4% a n d 4.2%, r espect ively; F igu r e 5A). St r a in ed ca r r ot pH dem on st r a t ed a
con sist en t r a t e of declin e, bu t spiked sa m ples exh ibit ed
a lower pH (5.28) du e t o a cidit y im pa r t ed by t h e
ph en olics a s com pa r ed t o con t r ol sa m ples (pH 5.82;
F igu r e 5B). Th is a cidifica t ion effect ser ved t o lower t h e
in it ia l declin e in pH a ft er pr ocessin g for t h e spiked
sa m ples (0.08 u n it ) a s com pa r ed t o t h e con t r ol (0.74
u n it ). Th is sm a ll declin e in pH a ft er pr ocessin g for
spiked sa m ples dem on st r a t ed t h a t eit h er oxida t ive
r ea ct ion s or a ch a n ge in t h e bu ffer in g a bilit y of t h e
pu r ee r esu lt ed in a pH declin e du r in g pr ocessin g. Sin ce
oxida t ion r a t es a r e gen er a lly pH depen den t , t h e r a t e of
pH declin e wa s slower for spiked sa m ples a s com pa r ed
t o t h e con t r ol du r in g t h e fir st week of st or a ge. It wa s
h ypot h esized t h a t in cr ea sin g ph en olic a cid con cen t r a t ion wou ld r esu lt in m or e r a pid losses of solu ble
ph en olics, r esu lt in g in a fa st er declin e in pH , a n d wou ld
r esu lt in r a pid discolor a t ion du r in g st or a ge. H owever ,
spiked sa m ples r esu lted in a 16.6% loss of ph en olic a cids
a s com pa r ed t o a 31.8% loss in n on spiked sa m ples.
Resu lt s in dica t e t h a t t h e r ea ct ion r a t e of ph ysicoch em ica l ch a n ges wou ld likely be con st a n t u n der sim ila r
st or a ge con dit ion s r ega r dless of ph en olic con cen t r a t ion ,
bu t a lower pu r ee pH wou ld ser ve t o slow oxida t ive

J. Agric. Food Chem., Vol. 47, No. 5, 1999 2113

F ig u re 4. E ffect of pr et r ea t m en t on m olecu la r weigh t dist r ibu t ion (kDa ) of ph en olic a cids in st r a in ed ca r r ot s a ft er 4 weeks
storage at 40 C. Treatments include no pretreatment (control),
n it r ogen spa r ge (N 2 spa r ge), bla n ch ed/fr ozen wit h n it r ogen
spa r ge (BF N 2 ), a n d oxygen spa r ge (O 2 spa r ge).
Ta ble 1. Co n c e n tra tio n o f Wa te r-S o lu ble P h e n o lic s
(m g /k g ) in S tra in e d Ca rro ts a fte r Th e rm a l P ro c e s s in g
(D a y 0) a n d a fte r 4 We e k s o f S to ra g e (We e k 4) in a n
Ac c e le ra te d S h e lf Life He ld a t 40 C fo r 4 We e k s a
t r ea t m en t

da y 0 (m g/kg)

week 4 (m g/kg

con t r ol
N2
BF N 2
O2

223
252
253
207

188
200
202
145

a A m olecu la r weigh t colu m n wa s u sed t o sepa r a t e ph en olic a cid

fr a ct ion s (m a x ) 280 n m ) a n d qu a n t ifica t ion wa s con du ct ed by
t h e F olin -Cioca lt eu a ssa y.

r ea ct ion s a ffect in g st r a in ed ca r r ot color . On t h e ba sis

of per cen t a ge loss, t h e effect of ph en olic a cid con cen t r a t ion a n d pH declin e wa s sim ila r bet ween spiked a n d
con t r ol sa m ples. In dividu a l ph en olic a cids h a ve been
sh own t o oxidize a t differ en t r a t es (Ga r cia et a l., 1992);
t h er efor e, con cen t r a t ion a n d iden t it y of t h ese com pounds after processing and storage may impact strained
ca r r ot color .
Mo d e l S y s te m . Ca r r ot m odel solu t ion s wer e pr epa r ed t o ver ify t h e m a jor r ole of ph en olic oxida t ion in
color pr odu ct ion a n d t o det er m in e if Ma illa r d br own in g
wa s a pot en t ia l sou r ce of pigm en t a t ion du r in g com m er cia l pr ocessin g a pplica t ion s. Aft er h ea t in g ea ch
m odel syst em , solu t ion pH wa s a dju st ed t o or igin a l
levels a n d color wa s m on it or ed a t 420 n m . Color
for m a t ion du e t o Ma illa r d br own in g wa s det er m in ed by
su bt r a ct in g t h e a bsor ba n ce du e t o ph en olic a cid con den sa t ion (n o cyst ein e a dded) fr om t h e t ot a l color
for m ed (wit h cyst ein e) a n d expr essed on a per cen t a ge
ba sis (Ta ble 2). P h en olic a cid con den sa t ion wa s r espon sible for t h e yellow a n d br own pigm en t s for m ed, wit h

2114 J. Agric. Food Chem., Vol. 47, No. 5, 1999

Talcott and Howard

com bin a t ion wit h dissolved oxygen is det r im en t a l t o

st r a in ed ca r r ot color , a n d a t t em pt s sh ou ld be m a de t o
r edu ce oxygen con cen t r a t ion in t h e syst em t o pr even t
ph en olic a u t oxida t ion a n d ca r ot en e blea ch in g.
LITE RATURE CITE D

F ig u re 5. E ffect s of pr et r ea t m en t on (A) t ot a l solu ble ph en olics a n d (B) pH in st r a in ed ca r r ot s st or ed 4 weeks a t 40 C.

Tr ea t m en t s in clu ded a n on spiked con t r ol a n d a ph en olic a cid
spike (1250 mg/kg). Unprocessed puree demonstrates changes
du r in g t h er m a l pr ocessin g. Ba r s r epr esen t st a n da r d er r or of
t h e m ea n .
Ta ble 2. Co n tribu tio n (% o f To ta l Co lo r F o rm a tio n ) o f
P h e n o lic Co n d e n s a tio n a n d Ma illa rd B ro w n in g a t 420
n m to To ta l Co lo r F o rm a tio n in a S tra in e d Ca rro t Mo d e l
S o lu tio n a
t ot a l br own in g
in it ia l pH

ph en olic br own in g (%)

Ma illa r d br own in g (%)

7.00
6.50
6.00
5.50
5.00

54.9
68.3
97.5
88.0
100

45.1
31.7
2.2
12.0
0

a P h en olic br own in g ca lcu la t ed a s color for m ed wit h ou t cyst ein e

m in u s color for m ed wit h cyst ein e. Model solu t ion con t a in ed 3%
su cr ose, 0.5% glu cose, 0.5% fr u ct ose, 50 m g/L ca ffeic a cid, 50 m g/L
syringic acid, 100 mg/L cysteine, and citric/malic acid (1:1) adjusted
t o pH 5-7. Solu t ion s wer e h ea t ed for 2 h a t 212 F .

lit t le con t r ibu t ion by Ma illa r d br own in g. Mor e pigm en t a t ion wa s for m ed a t h igh er pH va lu es a s r epor t ed
by n u m er ou s in vest iga t or s (Cer r u t t i et a l., 1985; Yeo
a n d Sh iba m ot o, 1991; F r iedm a n a n d Moln a r -P er l, 1990)
wh ich wer e a t t r ibu t a ble t o bot h ph en olic a cid con den sa t ion a n d Ma illa r d br own in g. Resu lt s in dica t ed t h a t color
for m a t ion du e t o Ma illa r d r ea ct ion s wou ld develop a t a
slow r a t e a t t h e pH r a n ge of st r a in ed ca r r ot s wit h t h e
majority of pigmentation (>88%) resulting from phenolic
con den sa t ion .
CONCLUSIONS

P r ocessor s sh ou ld a void u n it oper a t ion s t h a t a llow for

excessive h ea t a n d oxygen in cor por a t ion in t o m a cer a t ed
fr u it a n d veget a ble pr odu ct s du e t o ph en olic a u t oxida t ion pot en t ia l. Ut iliza t ion of n it r ogen spa r ge t ech n iqu es
im m edia t ely a ft er m a cer a t ion followed by h ea dspa ce
flu sh in g wit h n it r ogen pr ior t o ca ppin g sh ou ld im pr ove
over a ll color r et en t ion in st r a in ed ca r r ot s. H ea t in

Ar ch a n a , G. N.; P r a ka sh , J .; Ash a , M. R.; Ch a n d, N. E ffect s

of pr ocessin g on pigm en t s of cer t a in select ed veget a bles. J .
Food Qu al. 1995, 18, 91-101.
Ba bic, I.; Am iot , M. J .; Ngu yen -Th e, C.; Au ber t , S. Ch a n ges
in ph en olic con t en t in fr esh r ea dy-t o-u se sh r edded ca r r ot s
du r in g st or a ge. J . Food S ci. 1993, 58, 351-356.
Bu ch eli, C. S.; Robin son , S. P . Con t r ibu t ion s of en zym a t ic
br own in g t o color in su ga r ca n e ju ice. J . Agric. Food Ch em .
1994, 42, 257-261.
Cer r u t t i, P .; Resn ik, S. L.; Seldes, A.; F on t a n , C. F . Kin et ics
of det er ior a t e r ea ct ion s in m odel food syst em s of h igh wa t er
a ct ivit y: glu cose loss, 5-h ydr oxym et h ylfu r fu r a l a ccu m u la t ion a n d flu or escen ce developm en t du e t o n on en zym a t ic
br own in g. J . Food S ci. 1985, 50, 627-639, 656.
Ch eft el, J . C.; Cu q, J . L.; Lor ien t , D. Am in o a cids, pept ides,
a n d pr ot ein s. In Food Ch em istry; F en n em a , O. R., E d.;
Dekker : New Yor k, 1985; p 347.
Cillier s, J . J . L.; Sin glet on , V. L. Non en zym a t ic a u t oxida t ive
ph en olic br own in g r ea ct ion s in a ca ffeic a cid m odel syst em .
J . Agric. Food Ch em . 1989, 37, 890-896.
Cillier s, J . J . L.; Sin glet on , V. L. Ca ffeic a cid a u t oxida t ion a n d
t h e effect s of t h iols. J . Agric. Food Ch em . 1990, 38, 17891796.
Du Bois, M.; Gilles, K. A.; H a m ilt on , J . K.; Ren er s, P . A.; Sm it h ,
F . Color im et r ic m et h od for det er m in a t ion of su ga r s a n d
r ela t ed su bst a n ces. An al. Ch em . 1956, 28, 350-356.
F r iedm a n , M.; Moln a r -P er l, I. In h ibit ion of br own in g by su lfu r
a mino a cids. 1. Hea ted a mino a cid-glucose system s. J . Agric.
Food Ch em . 1990, 38, 1642-1647.
F u lcr a n d, H .; Ch em in a t , A.; Br ou illa r d, R.; Ch eyn ier , V.
Ch a r a ct er iza t ion of com pou n ds obt a in ed by ch em ica l oxida t ion of ca ffeic a cid in a cidic con dit ion s. Ph ytoch em istry 1994,
35, 499-505.
Ga r cia , P .; Br en es, M.; Va t t a n , T.; Ga r r ido, A. Kin et ic st u dy
a t differ en t pH va lu es of t h e oxida t ion pr ocess t o pr odu ce
r ipe olives. J . S ci. Food Agric. 1992, 60, 327-331.
Gr oss, J . Ca r ot en oids. In Pigm en ts in Vegetables: Ch loroph ylls
an d Caroten oid s; Va n Nost r a n d Rein h old: New Yor k, 1991;
p 100.
H owa r d, L. R.; Br a swell, D. D.; Asela ge, J . Ch em ica l com posit ion a n d color of st r a in ed ca r r ot s a s a ffect ed by pr ocessin g
J . Food S ci. 1996, 61, 327-330.
Ice, J . R.; Hamann, D. D.; Purcell, A. E. Effects of pH, enzymes,
a n d st or a ge t im e on t h e r h eology of sweet pot a t o pu r ee. J .
Food S ci. 1980, 45, 1614-1618.
Leon a r d, S.; P a n gbor n , R. M.; Tom br opou lo, D. In flu en ce of
processing variables on flavor stability of pear puree. J . Food
S ci. 1976, 41, 840-844.
Lit r idou , M.; Lin ssen , J .; Sch ols, H .; Ber gm a n s, M.; P ost h u m u s, M.; Tsim idou , M.; Boskou , D. P h en olic com pou n ds in
vir gin olive oils: fr a ct ion a t ion by solid-ph a se ext r a ct ion a n d
a n t ioxida n t a ct ivit y a ssessm en t . J . S ci. Food Agric. 1997,
74, 169-174.
Lu h , B. S.; An t on a kos, J .; Da ou d, H . N. Ch em ica l a n d qu a lit y
ch a n ges in st r a in ed ca r r ot s ca n n ed by t h e a sept ic a n d r et or t
pr ocess. Food T ech n ol. 1969, 23, 103-107.
Reyes, F . G. R.; P ooch a r oen , B.; Wr olst a d, R. E . A r esea r ch
n ot e. Ma illa r d br own in g of su ga r -glycin e m odel syst em s:
ch a n ges in su ga r con cen t r a t ion , color a n d a ppea r a n ce. J .
Food S ci. 1982, 47, 1376-1377.
Sa r ker , S. K.; P h a n , C. T. Na t u r a lly-occu r r in g a n d et h ylen ein du ced ph en olic com pou n ds in t h e ca r r ot r oot . J . Food
Protein 1979, 42, 526-534.
SAS In st it u t e, In c. S AS Users Gu id e: S tatistics, Version 5;
SAS In st it u t e: Ca r y, NC, 1985.
Sim on , P . W. Ca r r ot F la vor : effect s of gen ot ype, gr owin g
con dit ion s, st or a ge, a n d pr ocessin g. In E valu ation of Qu ality

Effect of Phenolic Acids on Carrot Puree Color

of Fru its an d Vegetables; E skin , E ., E d.; AVI P u blish in g
Co.: West por t , CT, 1985; pp 315-328.
Swa in , T.; H illis, W. E . Th e ph en olic con st it u en t s of Pru n u s
d om estica. I. Th e qu a n t it a t ive a n a lysis of ph en olic con st it u en t s. J . S ci. Food Agric. 1959, 10, 63-68.
Tor ibio, J . L.; Loza n o, J . E . H ea t in du ced br own in g of cla r ified
a pple ju ice a t h igh t em per a t u r es. J . Food S ci. 1986, 51,
172-175.
Ya n , R. F a ct or s a ffect in g ph en olic com pou n ds in ba by food
ca r r ot s. Ma st er s Th esis. Un iver sit y of Ar ka n sa s, F a yet t eville, AR, 1989.

J. Agric. Food Chem., Vol. 47, No. 5, 1999 2115

Ya n g, C. S. T.; Ya n g, P . P . A. E ffect of pH , cer t a in ch em ica ls
a n d h oldin g t im e-t em per a t u r e on t h e color of lowbu sh
blu eber r y pu r ee. J . Food S ci. 1987, 52, 346-352.
Yeo, H .; Sh iba m ot o, T. Micr owa ve-in du ced vola t iles of t h e
Ma illa r d m odel syst em u n der differ en t pH con dit ion s. J .
Agric. Food Ch em . 1991, 39, 370-373.
Received for r eview Oct ober 14, 1998. Revised m a n u scr ipt
r eceived Ma r ch 2, 1999. Accept ed Ma r ch 9, 1999.
J F 981134N