The Effect of Light and Temperature on the Stability of Anthocyanin Pigment from Hibiscus sabdariffa L
The Effect of Light and Temperature on the Stability of Anthocyanin
Pigment from Hibiscus sabdariffa L
Written by
Siti Nuryanti,\Sabiriii Matsjeh2, Chairil Anwar Jand Tri Joko Raharjo2
"
Student of Doctoral Programme, Departement of Chemistry, Faculty of Mathematics
and Natural Sciences, Gadjah MadaUniversity, Yogyakarta, Indonesia.
2> Departement of Chemistry, Faculty of Mathematics and Natural Sciences, Gadjah Mada
University, Yogyakarta, Indonesia
"
Corresponding author, tel :085241045745, email: sitinoer untad@vahoo.com
ABSTRACT
A research has been conducted to investigate the effect of light and temperature on
the stability of anthocyanin pigment from Hibiscus sabdariffa L. Flower Hibiscus
sabdariffa L was extracted sequentially by n-hexane, ethyl acetate, and methanol-acetic
acid 1% solvent. The anthocyanin pigment was detected in methanol-acetic acid 1%
extract. The anthocyanin pigment then was examined under various conditions of no
light' condition, 25 watt of lighting condition, with the temperature of 20° and 30° C. The
experiment showed that the feet about the presence of light and the increase of temperature
causing the anthocyanin pigment unstable.
i
Keywords: stability, anthocyanin, light, temperature, Hibiscus sabdariffa L
Introduction
Rosella (Hibiscus sabdariffa L) crown has not widely used commercially and only
discarded as waste, while the red calyx has been many utilized as food coloring agent and
healthy herbal tea. Researches on the utilization of Roselle calyx show that the calyx has
benefits in lowering cholesterol, blood pressure, blood sugar levels for diabetics
,
neutralizing toxins, inhibiting the cancer cells growth maintaining stamina, balancing body
,
weight, recovering from drugs addiction, reducing dizziness curing cough and diarrhea
,
(Chen et at, 2003; Farombi and Fakoya, 2005; Maganha et al, 2010) and as an anti oxidant
(Amor and Allaf, 2009).
Evidently, the petal contains compounds sucha ac, cyanidin-3-rutinoside and
delphinidin-3-glucoxyIoside as major anthocyanins and some others. Questions that arise
are
"
Does the crown of rosella have the same anthocyanin like the calyx T and "Do the
International Conference for Young Chemists (1CYC) 23Ki-25'k June 20I()\ "
At q.
'
J
/
f
light and temperature influence the stability of the crown anthocyanin?". Therefore,
Isolation and stability test of the crown anthocyanin toward the light and temperature are
urgently to be done. By conducting this research, the properties of the crown anthocyanin
could be obtained and the crown may be converted into commercial product like the petals,
thus increasing the economical value of the crown.
Research Methods
Materials
Rosella (Hibiscus sabdariffa L) crown which was collected from Sleman,
Yogyakarta, Indonesia and identified in the Plant Taxonomy Laboratory, Universitas
Gadjah Mada. The following chemicals were purchased from E.Merck with p.a grade.
There are methanol, ethanol, butanol, n-hexane, ethyl acetate, hydrochloric acid, acetic
acid, thin layer chrmatography and silica gel F 254Equipment
Laboratory glassware, black bottle, transparent bottle, extractor (IKA KS 130 basic),
Rotary evaporator Buchii (R-I24), pH meter (Hanna HI-8314), analytical mass balance
(Libror EB-330 Shimadzu), Fourier Transfrom Infra Red (FT-IT, Shimadzu 8201 PC),
Spectrophotometer UV-Vis (Miltonroy Array 3000), ,H-NMR spectrometer 500 MHz (Joel
JNM-MY 500, LIPI).
Procedures
v
Isolation ofAnthocyanin in Roselle Crown
As much as 100 g Rosella crown was washed, cut into small pieces and put into
evaporator flask. Then, extraction was performed using 500 mL of n-hexane for 20 hours
and filtered. The residue was extracted with 500 mL of ethyl acetate for 20 hours and
iltered, the extraction of residue using methanol-acetic acid 1% for 20 hours was
conducted and followed by filtration with Whattman filter paper. The red filtrate was
collected. Again, the residue was extracted with methanol-acetic acid 1% for 20 hours and
filtered and filtered. Extraction using methanol-acetic acid I%was stopped until the
°
colorless extract gained. The filtrate of filtering was collected and then evaporated at 65
C
International Conference for Young Chemists (ICYC) 2 3ÿ-2 5'h June 2010)
2
,
f
/
to
to obta,
obta,n
approx,mate,y 60 mL of concentrated f,,trate. The anthocyan,n was ana,yzed
us,ng UV-V,s
U
spectrophotometer at 200-700 nm and co,or test w,th Pb-acetate 1%,
Stability test of Rosetla crown anthocyanin toward light and temperature effects
As much as I mg of the isolated crystal was dissolved in 10 mL of methanol. The
solution was then divided into 2 parts (5 mL for each). The first part was inserted to the
transparent bottle and placed in the room which subjected to 25-watt light, whereas the
second was poured into the black bottle and placed in a black box. The temperature was set
to be 20°C. Next, 20 pL of each part was dissolved to 500 pL with methanol and analyzed
using spectrophotometer UV-Vis at X 200-700 nm. Absorbance of sample was observed
once every week (Laleh et al, 2006). The same procedure was performed at different
temperature, i.e. 30°C.
Results and Discussions
Identification of Rosella Crown Anthocyanin
The isolated Rosella crown using methanol-acetic acid 1% yielded red extract and
had maximum wavelenght at 536 nm. Harborne (1987) reported that anthocyanin that gave
maximum absorption at 535 nm in methanol-HCI solvent was cyanidin. To ensure that
,
two
color tests was done. First, the crown extract was exposed to ammonia vapor and gave
color change rom red to blue. Then, color test using Pb-acetate 1% reagent produced the
same color change like ammonia test, due to the formation of Pb-anthocyanin complex
.
According,
aSvjoig Jdtfv uoijooJ
i tfuapj
"(1003
pire BJjanbiA-BioiBo) ojnitjjodiuoi
treqi 3 03J° 3JnjBJ3duiaj je
Xq papgjjB XjSuojjs sbm XjijTqBjs uiireKooqjue aqj aouaq
o
J3MOJS SBM UIUBXooqjUB JO 3SB3J03p 3DUBqjOSqB 3qj 'UOIJippB U] (£003
J® BAO(ipBS) 30g5
jb SuxjBsq 01 jubjsis3j iBqj sjuatuSid /Cwaqjapja jo %os Xjuo sbm. aiaqi SuijBaq jo sxnoq £
'
joyy *«TUBXoo™jBÿÿÿJ3pÿ
cm jo XjijiqBjs aqj poouartjjui osjb SuijEoqjo ssjftep puB aunj
3m *(9003 'lP Ja aqnÿqiiwÿ
)ÿ
njBjaduiaj oSbjojs aqj Xq papajjB sbm j3mo|j umajsvaddtfj
'
jo uiuByCooqjire 3ÿ
(3pisouBjXdoon{g-jj-{XsouBiXdouuiBqj-»-o-M9)-0-£-U!P?"Kÿ3 J°
/
jojoo oqx *uiugOS X 1K uorjdxosqB jo uoipnpaj aqi jb UMoqs sb j3mojj tunouqjoqvjnw
.svj$rf
>
jo uoijBpBiSap 3qi paj jqgquns jeqj pajejs (9003) 1° *9 euuef
001
L
-
LViL
£¥VQ
ZLtQ
6SC0
Lb169
L£LQ9
9
S
t
00 i
UO1O
66tQ
£
I8'€fr
60S'0
0S'l6
8?1€l
esi'o
66'99
lfr'01
908 0
8t'frfr
£0S"'0
I
0
906X)
0
906'0
0
'
(%)
asueqjosqe
aseajosQ
(%)
UiU 9£§
\v doueqjosqy
doueqjosqe
aseajosQ
Z
vau 9£§
dÿiieqjoj
-
7 nffuTjpqos snostqm umojo
juauiSid uiireXooqjUB jo XjqiqBjs aqj uo (3o0£) ajnjBjaduiaj puB jq3ij aqj pajjg ,Z ajqei
-
3®®
' ..
."»
J' S®
3E®
"S®&
se©
fe©
-fnTpTrÿ
f vr;.
l
600
«3&
-?00
ffafnoae-tsr"©
Figure 2. Spectra UV-Vis of anthocyanin cyanidin-3-0-(6"-0-a-rhamnopyranosil-p
giocopyranoside) from crown rosella
SaisBfbTterE
Figure 3. Spectra UV-Vis after degradation of anthocyanin cyanidin-3-0(6"-0arhamnopyranosil-p-glocopyranoside) from crown rosella
The
degradation
processes
of
cyanidin-3-O-(6"-0-a-rMmnopyranosyI-|3-
glucopyranoside) from Rosella crown by light and temperature, most likely the same as
happened to cyanidin-3-glucosyde from elderberry. There were deglicosylation (to form
cyanidin), followed by the breaking of A ring (to yield phloroglucinaldehyde) and the
breaking of B ring (to give protocatheeuic acid) as shown in Figure 4.
International Conference for Young Chemists (1CYC) 23ÿ-25* June 2010)
6
Protocateehme acid
Figure. 4. Possible temperature degradation mechanism of anthocyanin cyanidin-3-0-(6"O-a-
rhamnopyranosil-p- glocopyranoside) from crown rosella
Conclusions
To sum up, it can be concluded that Roselle (Hibiscus sabdariffa L) crown contained
anthocyanin with sugar functional group. The anthocyanin was the same with Hippeastrum
"
anthocyanin, i.e. cyanidin-3-0-(6
-
0-a-rhamnopyranosyl-P-glucopyranoside) The effect of
.
light and temperature, evidently, affect the stability of the crown anthocyanin.
This research is a first step and research about the convertion of Roselle crown into
commercial product is urgently required.
Acknowledgement
The author would like to thank the Directorate General of Higher Education, Ministry of
National Education, Republic of Indonesia, which has provided funding of research through
Grants Doctoral Research Program, Universitas Gadjah Mada, with
ÿ
contract number.
LPPM-UGM/1212/2009, date May 15"12009.
References
Adji, F., Yaves, F. L,, Emmanuelle, M., Paul, L., Agustin. A., Georges, A. N. and Emile. M.
G 2008, Anthocyanin Characterization of Pilot Plant Water Extracts of Delanoc
regia Flowers , Molecules, 13, 1238-1245.
,
Amor, B. and Allaf, K., 2009, Impact of Texturing Using Instant Pressure Drop Treatment
Prior to Solvent Extraction of Anthocyanins from Malaysian Roselle {Hibiscus
sabdariffa), J. Food Chem., 115, 820-825.
International Conference for Young Chemists (ICYC) 23"'-25'h June 2010)
1
Byamukama RMonica, J., Bernard, K„ Namukobe, J. and Qyvind M, A., 2006,
Athocyanin from Flowers of Hippeastrum Cultivars, J. Sceintia Hortic., 109, 262C., Hsu, J, D., Wang, S. F,, Chiang, H. C., Yang, M.Y,, Kao, E. S,, Ho, Y, C. and
Wang, C. J., 2003, Hibiscus sabdariffa extract Inhibits the Development of
Atherosclerosis in Cholesterol-fed Rabbits, J. Agric & Food Chem., 51, 5465-5472
C
,
arombi, E, O, & Fokoya, A„ 2005, Free Radical Scavenging and Antigenotoxic Activities
of Natural Phenolic Compounds in Dried Flowers of Hibiscus sabdariffa L, J.
Molecular Nutrition and Food Reseach, 49, 1118-1120.
ircia-Viquera, C., Zafirilla, P., Abellan, P., Artes, F. and Tomas-Barberan, F. A„ 1999,
Color stability of Stawberry Jam as Affected by Cultivar and Strorage Temperatura,
J of Food science, 64(2), 243-247.
.
ÿrborne. J. B., 1958, Spectral Methods of Characterizing Anthocyanins, J. Biochem., 70,
22-28.
inna, O., KhairuL A., Maziah, M. and Mohd, Y., 2006, Flower Pigment Analysis of
Melastoma Malabathricum, J. Afr. BiotechnoL, 5 (2) 170-174.
,
\y
Iÿaleh, d H., Frydoonl~ar, R„ Heldary, R., Jamee and Zare, S., 2006, The Effect of Light,
Temperature, pH and Species on Stability of Anthocyanin Pigments in Four
Berber is Species, J. Nutr., 5 (1) 90-92.
j
/
JlAaganha, E. G„ Rafael da Costa, H„ Renato, M, R„ Joao, A.P, II,. Ana Ligia Lia de Paula,
R. and Jeifer, S., 2010, Pharmacological Evidences for the Extracts and Secondary
Metabolites from Plants of the Genus Hibisus-a Review, J. Food Chem., 118, 1-10.
j /Patras A„ Nigel P. B„ Colm O. D, and Tiwari B. K., 2010, Effect of Thermal Processing on
Anthocyanin Stability in Foods; Mechanisms and Kinetics of Degradation a
Rreview, J. Food Sci & Tech., 21 3-11
Ribereau-Gayon, P., 1969, Les Composess Phenoliquesdes Vegetaux Les Anthocyanin,
Dunod Paris, 142-147.
Sadilova, E., Carle, R. and Stintzing, F. C., 2007, Thermal Degradation of Anthocyanins
and its Impect on Color and In Vitro Antioxidant Capacity,./ of Food Scin, 71, 504512.
International Conference for Young Chemists (ICYC) 2 3rJ-2 ÿJune 2010)
8
Pigment from Hibiscus sabdariffa L
Written by
Siti Nuryanti,\Sabiriii Matsjeh2, Chairil Anwar Jand Tri Joko Raharjo2
"
Student of Doctoral Programme, Departement of Chemistry, Faculty of Mathematics
and Natural Sciences, Gadjah MadaUniversity, Yogyakarta, Indonesia.
2> Departement of Chemistry, Faculty of Mathematics and Natural Sciences, Gadjah Mada
University, Yogyakarta, Indonesia
"
Corresponding author, tel :085241045745, email: sitinoer untad@vahoo.com
ABSTRACT
A research has been conducted to investigate the effect of light and temperature on
the stability of anthocyanin pigment from Hibiscus sabdariffa L. Flower Hibiscus
sabdariffa L was extracted sequentially by n-hexane, ethyl acetate, and methanol-acetic
acid 1% solvent. The anthocyanin pigment was detected in methanol-acetic acid 1%
extract. The anthocyanin pigment then was examined under various conditions of no
light' condition, 25 watt of lighting condition, with the temperature of 20° and 30° C. The
experiment showed that the feet about the presence of light and the increase of temperature
causing the anthocyanin pigment unstable.
i
Keywords: stability, anthocyanin, light, temperature, Hibiscus sabdariffa L
Introduction
Rosella (Hibiscus sabdariffa L) crown has not widely used commercially and only
discarded as waste, while the red calyx has been many utilized as food coloring agent and
healthy herbal tea. Researches on the utilization of Roselle calyx show that the calyx has
benefits in lowering cholesterol, blood pressure, blood sugar levels for diabetics
,
neutralizing toxins, inhibiting the cancer cells growth maintaining stamina, balancing body
,
weight, recovering from drugs addiction, reducing dizziness curing cough and diarrhea
,
(Chen et at, 2003; Farombi and Fakoya, 2005; Maganha et al, 2010) and as an anti oxidant
(Amor and Allaf, 2009).
Evidently, the petal contains compounds sucha ac, cyanidin-3-rutinoside and
delphinidin-3-glucoxyIoside as major anthocyanins and some others. Questions that arise
are
"
Does the crown of rosella have the same anthocyanin like the calyx T and "Do the
International Conference for Young Chemists (1CYC) 23Ki-25'k June 20I()\ "
At q.
'
J
/
f
light and temperature influence the stability of the crown anthocyanin?". Therefore,
Isolation and stability test of the crown anthocyanin toward the light and temperature are
urgently to be done. By conducting this research, the properties of the crown anthocyanin
could be obtained and the crown may be converted into commercial product like the petals,
thus increasing the economical value of the crown.
Research Methods
Materials
Rosella (Hibiscus sabdariffa L) crown which was collected from Sleman,
Yogyakarta, Indonesia and identified in the Plant Taxonomy Laboratory, Universitas
Gadjah Mada. The following chemicals were purchased from E.Merck with p.a grade.
There are methanol, ethanol, butanol, n-hexane, ethyl acetate, hydrochloric acid, acetic
acid, thin layer chrmatography and silica gel F 254Equipment
Laboratory glassware, black bottle, transparent bottle, extractor (IKA KS 130 basic),
Rotary evaporator Buchii (R-I24), pH meter (Hanna HI-8314), analytical mass balance
(Libror EB-330 Shimadzu), Fourier Transfrom Infra Red (FT-IT, Shimadzu 8201 PC),
Spectrophotometer UV-Vis (Miltonroy Array 3000), ,H-NMR spectrometer 500 MHz (Joel
JNM-MY 500, LIPI).
Procedures
v
Isolation ofAnthocyanin in Roselle Crown
As much as 100 g Rosella crown was washed, cut into small pieces and put into
evaporator flask. Then, extraction was performed using 500 mL of n-hexane for 20 hours
and filtered. The residue was extracted with 500 mL of ethyl acetate for 20 hours and
iltered, the extraction of residue using methanol-acetic acid 1% for 20 hours was
conducted and followed by filtration with Whattman filter paper. The red filtrate was
collected. Again, the residue was extracted with methanol-acetic acid 1% for 20 hours and
filtered and filtered. Extraction using methanol-acetic acid I%was stopped until the
°
colorless extract gained. The filtrate of filtering was collected and then evaporated at 65
C
International Conference for Young Chemists (ICYC) 2 3ÿ-2 5'h June 2010)
2
,
f
/
to
to obta,
obta,n
approx,mate,y 60 mL of concentrated f,,trate. The anthocyan,n was ana,yzed
us,ng UV-V,s
U
spectrophotometer at 200-700 nm and co,or test w,th Pb-acetate 1%,
Stability test of Rosetla crown anthocyanin toward light and temperature effects
As much as I mg of the isolated crystal was dissolved in 10 mL of methanol. The
solution was then divided into 2 parts (5 mL for each). The first part was inserted to the
transparent bottle and placed in the room which subjected to 25-watt light, whereas the
second was poured into the black bottle and placed in a black box. The temperature was set
to be 20°C. Next, 20 pL of each part was dissolved to 500 pL with methanol and analyzed
using spectrophotometer UV-Vis at X 200-700 nm. Absorbance of sample was observed
once every week (Laleh et al, 2006). The same procedure was performed at different
temperature, i.e. 30°C.
Results and Discussions
Identification of Rosella Crown Anthocyanin
The isolated Rosella crown using methanol-acetic acid 1% yielded red extract and
had maximum wavelenght at 536 nm. Harborne (1987) reported that anthocyanin that gave
maximum absorption at 535 nm in methanol-HCI solvent was cyanidin. To ensure that
,
two
color tests was done. First, the crown extract was exposed to ammonia vapor and gave
color change rom red to blue. Then, color test using Pb-acetate 1% reagent produced the
same color change like ammonia test, due to the formation of Pb-anthocyanin complex
.
According,
aSvjoig Jdtfv uoijooJ
i tfuapj
"(1003
pire BJjanbiA-BioiBo) ojnitjjodiuoi
treqi 3 03J° 3JnjBJ3duiaj je
Xq papgjjB XjSuojjs sbm XjijTqBjs uiireKooqjue aqj aouaq
o
J3MOJS SBM UIUBXooqjUB JO 3SB3J03p 3DUBqjOSqB 3qj 'UOIJippB U] (£003
J® BAO(ipBS) 30g5
jb SuxjBsq 01 jubjsis3j iBqj sjuatuSid /Cwaqjapja jo %os Xjuo sbm. aiaqi SuijBaq jo sxnoq £
'
joyy *«TUBXoo™jBÿÿÿJ3pÿ
cm jo XjijiqBjs aqj poouartjjui osjb SuijEoqjo ssjftep puB aunj
3m *(9003 'lP Ja aqnÿqiiwÿ
)ÿ
njBjaduiaj oSbjojs aqj Xq papajjB sbm j3mo|j umajsvaddtfj
'
jo uiuByCooqjire 3ÿ
(3pisouBjXdoon{g-jj-{XsouBiXdouuiBqj-»-o-M9)-0-£-U!P?"Kÿ3 J°
/
jojoo oqx *uiugOS X 1K uorjdxosqB jo uoipnpaj aqi jb UMoqs sb j3mojj tunouqjoqvjnw
.svj$rf
>
jo uoijBpBiSap 3qi paj jqgquns jeqj pajejs (9003) 1° *9 euuef
001
L
-
LViL
£¥VQ
ZLtQ
6SC0
Lb169
L£LQ9
9
S
t
00 i
UO1O
66tQ
£
I8'€fr
60S'0
0S'l6
8?1€l
esi'o
66'99
lfr'01
908 0
8t'frfr
£0S"'0
I
0
906X)
0
906'0
0
'
(%)
asueqjosqe
aseajosQ
(%)
UiU 9£§
\v doueqjosqy
doueqjosqe
aseajosQ
Z
vau 9£§
dÿiieqjoj
-
7 nffuTjpqos snostqm umojo
juauiSid uiireXooqjUB jo XjqiqBjs aqj uo (3o0£) ajnjBjaduiaj puB jq3ij aqj pajjg ,Z ajqei
-
3®®
' ..
."»
J' S®
3E®
"S®&
se©
fe©
-fnTpTrÿ
f vr;.
l
600
«3&
-?00
ffafnoae-tsr"©
Figure 2. Spectra UV-Vis of anthocyanin cyanidin-3-0-(6"-0-a-rhamnopyranosil-p
giocopyranoside) from crown rosella
SaisBfbTterE
Figure 3. Spectra UV-Vis after degradation of anthocyanin cyanidin-3-0(6"-0arhamnopyranosil-p-glocopyranoside) from crown rosella
The
degradation
processes
of
cyanidin-3-O-(6"-0-a-rMmnopyranosyI-|3-
glucopyranoside) from Rosella crown by light and temperature, most likely the same as
happened to cyanidin-3-glucosyde from elderberry. There were deglicosylation (to form
cyanidin), followed by the breaking of A ring (to yield phloroglucinaldehyde) and the
breaking of B ring (to give protocatheeuic acid) as shown in Figure 4.
International Conference for Young Chemists (1CYC) 23ÿ-25* June 2010)
6
Protocateehme acid
Figure. 4. Possible temperature degradation mechanism of anthocyanin cyanidin-3-0-(6"O-a-
rhamnopyranosil-p- glocopyranoside) from crown rosella
Conclusions
To sum up, it can be concluded that Roselle (Hibiscus sabdariffa L) crown contained
anthocyanin with sugar functional group. The anthocyanin was the same with Hippeastrum
"
anthocyanin, i.e. cyanidin-3-0-(6
-
0-a-rhamnopyranosyl-P-glucopyranoside) The effect of
.
light and temperature, evidently, affect the stability of the crown anthocyanin.
This research is a first step and research about the convertion of Roselle crown into
commercial product is urgently required.
Acknowledgement
The author would like to thank the Directorate General of Higher Education, Ministry of
National Education, Republic of Indonesia, which has provided funding of research through
Grants Doctoral Research Program, Universitas Gadjah Mada, with
ÿ
contract number.
LPPM-UGM/1212/2009, date May 15"12009.
References
Adji, F., Yaves, F. L,, Emmanuelle, M., Paul, L., Agustin. A., Georges, A. N. and Emile. M.
G 2008, Anthocyanin Characterization of Pilot Plant Water Extracts of Delanoc
regia Flowers , Molecules, 13, 1238-1245.
,
Amor, B. and Allaf, K., 2009, Impact of Texturing Using Instant Pressure Drop Treatment
Prior to Solvent Extraction of Anthocyanins from Malaysian Roselle {Hibiscus
sabdariffa), J. Food Chem., 115, 820-825.
International Conference for Young Chemists (ICYC) 23"'-25'h June 2010)
1
Byamukama RMonica, J., Bernard, K„ Namukobe, J. and Qyvind M, A., 2006,
Athocyanin from Flowers of Hippeastrum Cultivars, J. Sceintia Hortic., 109, 262C., Hsu, J, D., Wang, S. F,, Chiang, H. C., Yang, M.Y,, Kao, E. S,, Ho, Y, C. and
Wang, C. J., 2003, Hibiscus sabdariffa extract Inhibits the Development of
Atherosclerosis in Cholesterol-fed Rabbits, J. Agric & Food Chem., 51, 5465-5472
C
,
arombi, E, O, & Fokoya, A„ 2005, Free Radical Scavenging and Antigenotoxic Activities
of Natural Phenolic Compounds in Dried Flowers of Hibiscus sabdariffa L, J.
Molecular Nutrition and Food Reseach, 49, 1118-1120.
ircia-Viquera, C., Zafirilla, P., Abellan, P., Artes, F. and Tomas-Barberan, F. A„ 1999,
Color stability of Stawberry Jam as Affected by Cultivar and Strorage Temperatura,
J of Food science, 64(2), 243-247.
.
ÿrborne. J. B., 1958, Spectral Methods of Characterizing Anthocyanins, J. Biochem., 70,
22-28.
inna, O., KhairuL A., Maziah, M. and Mohd, Y., 2006, Flower Pigment Analysis of
Melastoma Malabathricum, J. Afr. BiotechnoL, 5 (2) 170-174.
,
\y
Iÿaleh, d H., Frydoonl~ar, R„ Heldary, R., Jamee and Zare, S., 2006, The Effect of Light,
Temperature, pH and Species on Stability of Anthocyanin Pigments in Four
Berber is Species, J. Nutr., 5 (1) 90-92.
j
/
JlAaganha, E. G„ Rafael da Costa, H„ Renato, M, R„ Joao, A.P, II,. Ana Ligia Lia de Paula,
R. and Jeifer, S., 2010, Pharmacological Evidences for the Extracts and Secondary
Metabolites from Plants of the Genus Hibisus-a Review, J. Food Chem., 118, 1-10.
j /Patras A„ Nigel P. B„ Colm O. D, and Tiwari B. K., 2010, Effect of Thermal Processing on
Anthocyanin Stability in Foods; Mechanisms and Kinetics of Degradation a
Rreview, J. Food Sci & Tech., 21 3-11
Ribereau-Gayon, P., 1969, Les Composess Phenoliquesdes Vegetaux Les Anthocyanin,
Dunod Paris, 142-147.
Sadilova, E., Carle, R. and Stintzing, F. C., 2007, Thermal Degradation of Anthocyanins
and its Impect on Color and In Vitro Antioxidant Capacity,./ of Food Scin, 71, 504512.
International Conference for Young Chemists (ICYC) 2 3rJ-2 ÿJune 2010)
8