Anthocyanin Complex: Characterization and Cytotoxicity Studies
Complexation of anthocyanins to mimic natural
copigmentation process was investigated. Cyanidin-rich extracts from
Zea mays L. ceritina Kulesh. and delphinidin-rich extracts from
Clitoria ternatea L. were used to form 4 anthocyanin complexes,
AC1, AC2, AC3 and AC4, in the presence of several polyphenols and
a trace metal. Characterizations of the ACs were conducted by UV,
FTIR, DSC/TGA and morphological observations. Bathochromic
shifts of the UV spectra of 4 formulas of ACs were observed at peak
wavelengths of about 510-620 nm by 10 nm suggesting complex
formation. FTIR spectra of the ACs indicate shifts of peaks from
1,733 cm-1 to 1,696 cm-1 indicating interactions and a decrease in the
peak areas within the wavenumber of 3,400-3,500 cm-1 indicating
changes in hydrogen bonding. Thermal analysis of all of the ACs
suggests increases in melting temperature after complexation. AC
with the highest melting temperature was morphologically observed
by SEM and TEM to be crystal-like particles within a range of 50 to
200 nm. Particle size analysis of the AC by laser diffraction gave a
range of 50-600 nm, indicating aggregation. This AC was shown to
have no cytotoxic effect on cultured HGEPp0.5 and HGF (all p>
0.05) by MTT. Therefore, complexation of anthocyanins was simple
and self-assembly process, potentially resulting in nanosized particles
of anthocyanin complex.
[1] E. Pojer, F. Mattivi, D. Johnson, C. S. Stockley, “The Case for
Anthocyanin Consumption to Promote Human Health: A Review”
Compr.Rev. Food Sci. F. vol. 12, pp. 483, 2013.
[2] M. Shiono, N. Matsugaki, K. Takeda, “Phytochemistry: structure of the
blue cornflower pigment” Nature vol. 436, pp. 791, 2005.
[3] Y. Zhu et al., “Anti-inflammatory effect of purified dietary anthocyanin
in adults with hypercholesterolemia: A randomized controlled trial”
Nutr. Metab. Cardiovasc. Dis. vol. 23, pp. 843, 2013.
[4] J. Y. Choi et al., “Analysis and tentative structure elucidation of new
anthocyanins in fruit peel of Vitiscoignetiae Pulliat (meoru) using LCMS/
MS: Contribution to the overall antioxidant activity” J. Sep. Sci. vol.
33, pp. 1192, 2010.
[5] C. Ubeda et al., “Employment of different processes for the production
of strawberry vinegars: Effects on antioxidant activity, total phenols and
monomeric anthocyanins” LWT - Food Sci. Technol. vol. 52, pp. 139,
2013.
[6] S. C. Thomasset, University of Leicester (2008).
[7] L. C. Vasconcelos, M. C. Sampaio, F. C. Sampaio, J. S. Higino, “Use of
Punicagranatum as an antifungal agent against candidosis associated
with denture stomatitis” Mycoses vol. 46, pp. 192, 2003.
[8] A. Cisowska, D. Wojnicz, A. Hendrich, “Anthocyanins as antimicrobial
agents of natural plant origin.” Nat. Prod. Commun.vol. 6, pp. 149,
2011.
[9] Z. Yang, Y. Han, Z. Gu, G. Fan, Z. Chen, “Thermal degradation kinetics
of aqueous anthocyanins and visual color of purple corn (Zea mays L.)
cob” Innov. Food Sci. Emerg. Technol. vol. 9, pp. 341, 2008.
[10] Z. Yang, W. Zhai, “Identification and antioxidant activity of
anthocyanins extracted from the seed and cob of purple corn (Zea mays
L.)”Innov. Food Sci. Emerg. Technol. vol. 11, pp. 169, 2010.
[11] K. Kazuma, N. Noda, M. Suzuki, “Flavonoid composition related to
petal color in different lines of Clitoria ternatea” Phytochemistry vol. 64,
pp. 1133, 2003.
[12] M. Ponec, “In vitro cultured human skin cells as alternatives to animals
for skin irritancy screening” Int.J.Cosmet.Sci. vol. 14, pp. 245, 1992.
[13] R. K. Ward, A. W. Hubbard, H. Sulley, M. J. Garle, R. H. Clothier, “
Human keratinocyte cultures in an in vitro approach for the assessment
of surfactant-induced irritation” Toxicol.in Vitro vol. 12, pp. 163, 1998.
[14] H. Wang et al., “Antioxidant and anti-inflammatory activities of
anthocyanins and their aglycon, cyanidin, from tart cherries” J.
Nat.Prod.vol. 62, pp. 294, 1999.
[15] I. T. Nizamutdinova et al., “Anthocyanins from black soybean seed
coats preferentially Inhibit TNF-α-mediated Induction of VCAM-1 over
ICAM-1 through the regulation of GATAs and IRF-1” J. Agri. Food
Chem.vol. 57, pp. 7324, 2009.
[16] F. Afaq et al., “Delphinidin, an anthocyanidin in pigmented fruits and
vegetables, protects human HaCa T keratinocytes and mouse skin
against UVB-mediated oxidative stress and apoptosis” J. Invest.
Dermatol. vol. 127, pp. 222, 2007.
[17] D. X. Hou, X. Tong, N. Terahara, D. Luo, M. Fujii, “Delphinidin 3-
sambubioside, a Hibiscus anthocyanin, induces apoptosis in human
leukemia cells through reactive oxygen species-mediated mitochondrial
pathway” Arch. Biochem. Biophys. vol. 440 pp. 101, 2005.
[18] P.H. Shih, C.T. Yeh, G.C. Yen, “Anthocyanins induce the activation of
phase II enzymes through the antioxidant response element pathway
against oxidative stress-induced apoptosis” J. Agri. Food Chem.vol. 55,
pp. 9427, 2007.
[19] K. Tsoyi et al., “Protective effect of anthocyanins from black soybean
seed coats on uvb-induced apoptotic cell death in vitro and in vivo” J.
Agri. Food Chem.vol. 56, pp. 10600, 2008.
[20] S. R. Mallery et al., “Formulation and in-vitro and in-vivo evaluation of
a mucoadhesive gel containing freeze dried black raspberries:
implications for oral cancer chemoprevention” Pharm. Res. vol. 24, pp.
728, 2007.
[21] B. S. Shumway et al., “Effects of a topically applied bioadhesive berry
gel on loss of heterozygosity indices in premalignant oral lesions” Clin.
Cancer Res. vol. 14, pp. 2421, 2008.
[22] R. Boulton, “The copigmentation of anthocyanins and its role in the
color of red wine: a critical review” Am. J. Enol. Vitic. vol. 52, pp. 67,
2001.
[23] Y. Osawa, Copigmentation of anthocyanins. In: anthocyanins as food
colors. P. Markakis, Ed., (Academic Press Inc., New York, 1982).
[24] M. Rein, University of Helsinki (2005).
[25] M. S. Starr, F. J. Francis, “Effect of metallic ions on color and pigment
content of cranberry juice cocktail” J. Food Sci. vol. 38, pp. 1043, 1974.
[26] J. Sun, X. Cao, W. Bai, X. Liao, X. Hu, “Comparative analyses of
copigmentation of cyanidin 3-glucoside and cyanidin 3-sophoroside
from red raspberry fruits” Food Chem. vol. 120, pp. 1131, 2010.
[27] C. H. Brenes, D. D. Pozo-Insfran, S. T. Talcott, “Stability of
copigmented anthocyanins and ascorbic acid in a grape juice model
system” J. Agri. Food Chem. vol. 53, pp. 49, 2005.
[28] E. F. Gris, E. A. Ferreira, L. D. Falca˜o, M. T. Bordignon-Luiz, “Caffeic
acid copigmentation of anthocyanins from Cabernet Sauvignon grape
extracts in model systems” Food Chem. vol. 100, pp. 1289, 2007.
[29] M. Kopjar, V. Piližota, “Copigmentation effect of phenolic compounds
on red currant juice anthocyanins during storage” Croatian J. Food Sci.
Technol.vol. 1, pp. 16, 2009.
[30] S. Kunsági-Máté, E. Ortmann, L. Kollár, K. Szabó, M. P. Nikfardjam,
“Effect of ferrous and ferric ions on copigmentation in model solutions”
J. Mol. Struct. vol. 891, pp. 471, 2008.
[31] C. Zhang, Y. Ma, X. Zhao, J. Mu, “Influence of copigmentation on
stability of anthocyanins from purple potato peel in both liquid state and
solid state” J. Agri. Food Chem.vol. 57, pp. 9503, 2009.
[32] K. Torskangerpoll, Ø. M. Andersen, “Colour stability of anthocyanins in
aqueous solutions at various pH values” Food Chem. vol. 89, pp. 427,
2005. [33] M. A. Eskan, G. Hajishengallis, D. F. Kinane, “Differential activation of
human gingival epithelial cells and monocytes by
porphyromonasgingivalis fimbriae” Infect. Immun. vol. 75, pp. 892,
2007.
[34] A. Beklen, T. Sorsa, Y. T. Konttinen, “Toll-like receptors 2 and 5 in
human gingival epithelial cells co-operate with T-cell cytokine
interleukin-17” Oral Microbiol. Immunol. vol. 24, pp. 38, 2009.
[35] J. I. Gallin, R. Snyderman, D. T. Fearon, B. F. Haynes, C. Nathan,
Inflammation: Basic Principles and Clinical Correlates.(Lippincott
Williams & Wilkins, Philadelphia, ed. 3, 1999).
[36] T. Ara et al., “Human gingival fibroblasts are critical in sustaining
inflammation in periodontal disease” J. Periodontal Res. vol. 44, pp. 21,
2009.
[37] S. Singhabutta, 200 types of Thai herbal activities by
SountareeSinghabutta. (Koon, Bangkok, 1993).
[38] B. Khampaenjiraroch, A. Priprem, K. Lertrat, T. Damrongrungruang,
“Rapid HPLC of cyanidin and delphinidin of an anthocyanin complex
exposed to human gingival epithelial cells” Appl. Mech. Mater.vol. 563,
pp. 403, 2014.
[39] G. T. Sigurdson, M. M. Giusti, “Bathochromic and hyperchromic effects
of aluminum salt complexation by anthocyanins from edible sources for
blue color development” J. Agri. Food Chem.vol. 62, pp. 6955, 2014.
[40] C. A. Sims, J. R. Morris, “Effect of acetaldehyde and Tannins on the
color and chemical age of red muscadine (vitisrotundifolia) wine” Enol.
Vitic. vol. 37, pp. 163, 1986.
[1] E. Pojer, F. Mattivi, D. Johnson, C. S. Stockley, “The Case for
Anthocyanin Consumption to Promote Human Health: A Review”
Compr.Rev. Food Sci. F. vol. 12, pp. 483, 2013.
[2] M. Shiono, N. Matsugaki, K. Takeda, “Phytochemistry: structure of the
blue cornflower pigment” Nature vol. 436, pp. 791, 2005.
[3] Y. Zhu et al., “Anti-inflammatory effect of purified dietary anthocyanin
in adults with hypercholesterolemia: A randomized controlled trial”
Nutr. Metab. Cardiovasc. Dis. vol. 23, pp. 843, 2013.
[4] J. Y. Choi et al., “Analysis and tentative structure elucidation of new
anthocyanins in fruit peel of Vitiscoignetiae Pulliat (meoru) using LCMS/
MS: Contribution to the overall antioxidant activity” J. Sep. Sci. vol.
33, pp. 1192, 2010.
[5] C. Ubeda et al., “Employment of different processes for the production
of strawberry vinegars: Effects on antioxidant activity, total phenols and
monomeric anthocyanins” LWT - Food Sci. Technol. vol. 52, pp. 139,
2013.
[6] S. C. Thomasset, University of Leicester (2008).
[7] L. C. Vasconcelos, M. C. Sampaio, F. C. Sampaio, J. S. Higino, “Use of
Punicagranatum as an antifungal agent against candidosis associated
with denture stomatitis” Mycoses vol. 46, pp. 192, 2003.
[8] A. Cisowska, D. Wojnicz, A. Hendrich, “Anthocyanins as antimicrobial
agents of natural plant origin.” Nat. Prod. Commun.vol. 6, pp. 149,
2011.
[9] Z. Yang, Y. Han, Z. Gu, G. Fan, Z. Chen, “Thermal degradation kinetics
of aqueous anthocyanins and visual color of purple corn (Zea mays L.)
cob” Innov. Food Sci. Emerg. Technol. vol. 9, pp. 341, 2008.
[10] Z. Yang, W. Zhai, “Identification and antioxidant activity of
anthocyanins extracted from the seed and cob of purple corn (Zea mays
L.)”Innov. Food Sci. Emerg. Technol. vol. 11, pp. 169, 2010.
[11] K. Kazuma, N. Noda, M. Suzuki, “Flavonoid composition related to
petal color in different lines of Clitoria ternatea” Phytochemistry vol. 64,
pp. 1133, 2003.
[12] M. Ponec, “In vitro cultured human skin cells as alternatives to animals
for skin irritancy screening” Int.J.Cosmet.Sci. vol. 14, pp. 245, 1992.
[13] R. K. Ward, A. W. Hubbard, H. Sulley, M. J. Garle, R. H. Clothier, “
Human keratinocyte cultures in an in vitro approach for the assessment
of surfactant-induced irritation” Toxicol.in Vitro vol. 12, pp. 163, 1998.
[14] H. Wang et al., “Antioxidant and anti-inflammatory activities of
anthocyanins and their aglycon, cyanidin, from tart cherries” J.
Nat.Prod.vol. 62, pp. 294, 1999.
[15] I. T. Nizamutdinova et al., “Anthocyanins from black soybean seed
coats preferentially Inhibit TNF-α-mediated Induction of VCAM-1 over
ICAM-1 through the regulation of GATAs and IRF-1” J. Agri. Food
Chem.vol. 57, pp. 7324, 2009.
[16] F. Afaq et al., “Delphinidin, an anthocyanidin in pigmented fruits and
vegetables, protects human HaCa T keratinocytes and mouse skin
against UVB-mediated oxidative stress and apoptosis” J. Invest.
Dermatol. vol. 127, pp. 222, 2007.
[17] D. X. Hou, X. Tong, N. Terahara, D. Luo, M. Fujii, “Delphinidin 3-
sambubioside, a Hibiscus anthocyanin, induces apoptosis in human
leukemia cells through reactive oxygen species-mediated mitochondrial
pathway” Arch. Biochem. Biophys. vol. 440 pp. 101, 2005.
[18] P.H. Shih, C.T. Yeh, G.C. Yen, “Anthocyanins induce the activation of
phase II enzymes through the antioxidant response element pathway
against oxidative stress-induced apoptosis” J. Agri. Food Chem.vol. 55,
pp. 9427, 2007.
[19] K. Tsoyi et al., “Protective effect of anthocyanins from black soybean
seed coats on uvb-induced apoptotic cell death in vitro and in vivo” J.
Agri. Food Chem.vol. 56, pp. 10600, 2008.
[20] S. R. Mallery et al., “Formulation and in-vitro and in-vivo evaluation of
a mucoadhesive gel containing freeze dried black raspberries:
implications for oral cancer chemoprevention” Pharm. Res. vol. 24, pp.
728, 2007.
[21] B. S. Shumway et al., “Effects of a topically applied bioadhesive berry
gel on loss of heterozygosity indices in premalignant oral lesions” Clin.
Cancer Res. vol. 14, pp. 2421, 2008.
[22] R. Boulton, “The copigmentation of anthocyanins and its role in the
color of red wine: a critical review” Am. J. Enol. Vitic. vol. 52, pp. 67,
2001.
[23] Y. Osawa, Copigmentation of anthocyanins. In: anthocyanins as food
colors. P. Markakis, Ed., (Academic Press Inc., New York, 1982).
[24] M. Rein, University of Helsinki (2005).
[25] M. S. Starr, F. J. Francis, “Effect of metallic ions on color and pigment
content of cranberry juice cocktail” J. Food Sci. vol. 38, pp. 1043, 1974.
[26] J. Sun, X. Cao, W. Bai, X. Liao, X. Hu, “Comparative analyses of
copigmentation of cyanidin 3-glucoside and cyanidin 3-sophoroside
from red raspberry fruits” Food Chem. vol. 120, pp. 1131, 2010.
[27] C. H. Brenes, D. D. Pozo-Insfran, S. T. Talcott, “Stability of
copigmented anthocyanins and ascorbic acid in a grape juice model
system” J. Agri. Food Chem. vol. 53, pp. 49, 2005.
[28] E. F. Gris, E. A. Ferreira, L. D. Falca˜o, M. T. Bordignon-Luiz, “Caffeic
acid copigmentation of anthocyanins from Cabernet Sauvignon grape
extracts in model systems” Food Chem. vol. 100, pp. 1289, 2007.
[29] M. Kopjar, V. Piližota, “Copigmentation effect of phenolic compounds
on red currant juice anthocyanins during storage” Croatian J. Food Sci.
Technol.vol. 1, pp. 16, 2009.
[30] S. Kunsági-Máté, E. Ortmann, L. Kollár, K. Szabó, M. P. Nikfardjam,
“Effect of ferrous and ferric ions on copigmentation in model solutions”
J. Mol. Struct. vol. 891, pp. 471, 2008.
[31] C. Zhang, Y. Ma, X. Zhao, J. Mu, “Influence of copigmentation on
stability of anthocyanins from purple potato peel in both liquid state and
solid state” J. Agri. Food Chem.vol. 57, pp. 9503, 2009.
[32] K. Torskangerpoll, Ø. M. Andersen, “Colour stability of anthocyanins in
aqueous solutions at various pH values” Food Chem. vol. 89, pp. 427,
2005. [33] M. A. Eskan, G. Hajishengallis, D. F. Kinane, “Differential activation of
human gingival epithelial cells and monocytes by
porphyromonasgingivalis fimbriae” Infect. Immun. vol. 75, pp. 892,
2007.
[34] A. Beklen, T. Sorsa, Y. T. Konttinen, “Toll-like receptors 2 and 5 in
human gingival epithelial cells co-operate with T-cell cytokine
interleukin-17” Oral Microbiol. Immunol. vol. 24, pp. 38, 2009.
[35] J. I. Gallin, R. Snyderman, D. T. Fearon, B. F. Haynes, C. Nathan,
Inflammation: Basic Principles and Clinical Correlates.(Lippincott
Williams & Wilkins, Philadelphia, ed. 3, 1999).
[36] T. Ara et al., “Human gingival fibroblasts are critical in sustaining
inflammation in periodontal disease” J. Periodontal Res. vol. 44, pp. 21,
2009.
[37] S. Singhabutta, 200 types of Thai herbal activities by
SountareeSinghabutta. (Koon, Bangkok, 1993).
[38] B. Khampaenjiraroch, A. Priprem, K. Lertrat, T. Damrongrungruang,
“Rapid HPLC of cyanidin and delphinidin of an anthocyanin complex
exposed to human gingival epithelial cells” Appl. Mech. Mater.vol. 563,
pp. 403, 2014.
[39] G. T. Sigurdson, M. M. Giusti, “Bathochromic and hyperchromic effects
of aluminum salt complexation by anthocyanins from edible sources for
blue color development” J. Agri. Food Chem.vol. 62, pp. 6955, 2014.
[40] C. A. Sims, J. R. Morris, “Effect of acetaldehyde and Tannins on the
color and chemical age of red muscadine (vitisrotundifolia) wine” Enol.
Vitic. vol. 37, pp. 163, 1986.
@article{"International Journal of Biological, Life and Agricultural Sciences:68956", author = "Sucharat Limsitthichaikoon and Kedsarin Saodaeng and Aroonsri Priprem and Teerasak Damrongrungruang", title = "Anthocyanin Complex: Characterization and Cytotoxicity Studies", abstract = "Complexation of anthocyanins to mimic natural
copigmentation process was investigated. Cyanidin-rich extracts from
Zea mays L. ceritina Kulesh. and delphinidin-rich extracts from
Clitoria ternatea L. were used to form 4 anthocyanin complexes,
AC1, AC2, AC3 and AC4, in the presence of several polyphenols and
a trace metal. Characterizations of the ACs were conducted by UV,
FTIR, DSC/TGA and morphological observations. Bathochromic
shifts of the UV spectra of 4 formulas of ACs were observed at peak
wavelengths of about 510-620 nm by 10 nm suggesting complex
formation. FTIR spectra of the ACs indicate shifts of peaks from
1,733 cm-1 to 1,696 cm-1 indicating interactions and a decrease in the
peak areas within the wavenumber of 3,400-3,500 cm-1 indicating
changes in hydrogen bonding. Thermal analysis of all of the ACs
suggests increases in melting temperature after complexation. AC
with the highest melting temperature was morphologically observed
by SEM and TEM to be crystal-like particles within a range of 50 to
200 nm. Particle size analysis of the AC by laser diffraction gave a
range of 50-600 nm, indicating aggregation. This AC was shown to
have no cytotoxic effect on cultured HGEPp0.5 and HGF (all p>
0.05) by MTT. Therefore, complexation of anthocyanins was simple
and self-assembly process, potentially resulting in nanosized particles
of anthocyanin complex.
", keywords = "Anthocyanins, complexation, purple corn cops,
butterfly pea, physicochemical characteristics, cytotoxicity.", volume = "9", number = "2", pages = "147-7", }
