Volume 91, Issue 5, May (2004), pp. 757-764 © The Author 2004
doi:10.1079/BJN20041094

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Use of Tween 40 and Tween 80 to deliver a mixture of phytochemicals to human colonic adenocarcinoma cell (CaCo-2) monolayers

Sinead M. O'Sullivan¹, Julie A. Woods¹ and Nora M. O'Brien^2
1Department of Food and Nutritional Sciences, University College, Cork, Republic of Ireland
²Photobiology Unit, University of Dundee, Ninewells Hospital, Dundee, DD1 9SY, Scotland, UK

 (Received 15 September 2003–Revised 12 December 2003–Accepted 7 January 2004)

Epidemiological evidence suggests that dietary intake of carotenoids and tocopherols may influence the risk of certain chronic diseases, such as cancer and CVD. In vitro studies investigating the synergistic effects of mixtures of carotenoids and tocopherols have been hindered due to the difficulty of solubilising these lipophilic compounds. The objective of the present study was to develop a system for delivering tocopherols and carotenoids simultaneously to cells in culture. Differentiated human colonic adenocarcinoma cells (CaCo-2) were incubated with a mixture of these phytochemicals for 24h. The phytochemical mixture included carotenoids (astaxanthin, canthaxanthin, lutein, lycopene, α-carotene, β-carotene) and tocopherols (α-tocopherol and γ-tocopherol). The emulsifiers polyoxyethylene sorbitan monopalmitate (Tween 40) and polyoxyethylene sorbitan monooleate (Tween 80) were employed as the delivery vehicles, and were compared with tetrahydrofuran (THF). Each vehicle was added at a maximum concentration of 1ml/l. No toxic effects to the CaCo-2 cells were noted when Tween 40 or Tween 80 were used. Both Tween 40 and Tween 80 resulted in greater solubility of the mixture and delivered substantially more carotenoids and tocopherols to the cells than THF. In particular, lycopene was detected within the cells when Tween 40 and Tween 80 were employed, whereas it was below the limits of detection by HPLC when THF was used as the delivery vehicle. The phytochemicals were retained within the cells for 24h after supplementation. Tween 40 and Tween 80 have potential as simple, rapid and non-toxic methods for delivering mixtures of carotenoids and tocopherols to cells in culture.

Keywords:
Carotenoids, Polyoxyethylene sorbitan esters, Tween, Tetrahydrofuran, CaCo-2 cells

Abbreviations:
CAT, catalase, LDH, lactate dehydrogenase, MTT, 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide, NRUA, Neutral Red uptake assay, SOD, superoxide dismutase, THF, tetrahydrofuran