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ARS Home » Southeast Area » Little Rock, Arkansas » Arkansas Children's Nutrition Center » Research » Publications at this Location » Publication #195539

Title: ANTHOCYANIN (ACN) STABILITY IN CELL CULTURE MEDIA

Author
item WU, XIANLI - ACNC/UAMS
item Prior, Ronald
item BURT, T - INDIANA UNIVERSITY
item BELL, D - INDIANA UNIVERSITY

Submitted to: Journal of Federation of American Societies for Experimental Biology
Publication Type: Abstract Only
Publication Acceptance Date: 2/15/2006
Publication Date: 3/26/2006
Citation: Wu, X., Prior, R.L., Burt, T., Bell, D. 2006. Anthocyanin (ACN) stability in cell culture media [abstract]. Experimental Biology Annual Meeting, April, 1-5, 2006, San Francisco, CA. Paper No. LB377. Available: http://www.faseb.org/meetings/eb2006/call/default.htm.

Interpretive Summary: Anthocyanins (ACN's) are potential oxygen radical scavengers that have coronary vasoactive and vasoprotective properties. Cell or tissue culture systems have been used to examine the bioactivity and mechanisms of action of ACN's on the vascular system. However, due to their unique chemical structure, the stability of ACN's is a major concern in studies of this nature. In this study, three ACN enhanced extracts prepared from chokeberry (CB), bilberry (BB) and elderberry (EB) (10 mg/L) were added in Dulbecco’s Modified Eagles Media (HEPES buffered and phenol red free with bicarbonate) to evaluate their stability after 1h, 2h, 4h, 8h, 16h and 24h incubation at 37°C. Identification and quantification of ACN was carried out using a reverse phase HPLC-ESI/MS/MS system. ACN's were unstable in this media. The half life of most ACN's tested was less than 5h. However, different ACN's exhibited differences in stability. Aglycone seemed to be the major factor determining the stability for ACN's with similar sugar patterns. Delphinidin ACN's were least stable with a half life of only 0.43 ± 0.01 h (n=3), followed by petunidin (2.34 ± 0.04 h, n=3); malvidin (4.04 h, n=1); penonidin (4.31 h, n=2); and cyanidin (4.45 ± 0.60 h, n=10). A complex ACN (cyanidin 3-sambubioside-5-glucoside) was more stable than cyanidin 3-glucoside (8.76 h vs 4.05 h). Results from this study indicate that caution should be used in attributing changes in bioactivity measured in culture to the original ACN's, when degradation products could be responsible for the observed responses particularly when extended incubation times are used.

Technical Abstract: Anthocyanins (ACNs) are potential oxygen radical scavengers that have coronary vasoactive and vasoprotective properties. Cell or tissue culture systems have been used to examine the bioactivity and mechanisms of action of ACNs on the vascular system. However, due to their unique chemical structure, the stability of ACNs is a major concern in studies of this nature. In this study, three ACN enhanced extracts prepared from chokeberry (CB), bilberry (BB) and Elderberry (EB) (10 mg/L) were added in Dulbecco’s Modified Eagles Media (HEPES buffered and phenol red free with bicarbonate) to evaluate their stability after 1h, 2h, 4h, 8h, 16h and 24h incubation at 37°C. Identification and quantification of ACN was carried out using a reverse phase HPLC-ESI/MS/MS system. ACNs were unstable in this media. The half life of most ACNs tested was less than 5 h. However, different ACNS exhibited differences in stability. Aglycone seemed to be the major factor determining the stability for ACNs with similar sugar patterns. Delphinidin ACNs were least stable with a half life of only 0.43±0.01 h (n=3), followed by petunidin (2.34±0.04 h, n=3); malvidin (4.04 h, n=1); penonidin (4.31 h, n=2); and cyanidin (4.45±0.60 h, n=10). A complex ACN (cyanidin 3-sambubioside-5-glucoside) was more stable than cyanidin 3-glucoside (8.76 h vs 4.05 h). Results from this study indicate that caution should be used in attributing changes in bioactivity measured in culture to the original ACNs, when degradation products could be responsible for the observed responses particularly when extended incubation times are used.