Spectrochimica Acta Part A 64 (2006) 1002–1009 UV/vis, 1 H, and 13 C NMR spectroscopic studies to determine mangiferin pK a values Berenice G ´ omez-Zaleta a , Mar´ ıa Teresa Ram´ ırez-Silva a , Atilano Guti´ errez a , Enrique Gonz´ alez-Vergara b , Marisol G ¨ uizado-Rodr´ ıguez c , Alberto Rojas-Hern´ andez a, a Universidad Aut´ onoma Metropolitana-Iztapalapa, Depto. Qu´ ımica, ´ Area de Qu´ ımica Anal´ ıtica, Apdo. Postal 55-534, 09340 M´ exico DF, M´ exico b Centro de Qu´ ımica, Instituto de Ciencias, Benem´ erita Universidad Aut´ onoma de Puebla, M´ exico c Depto. de Qu´ ımica Inorg´ anica, Instituto de Qu´ ımica, UNAM, Ciudad Universitaria, M´ exico DF, M´ exico Received 14 June 2005; accepted 12 September 2005 Abstract The acid constants of mangiferin (a natural xanthonoid) in aqueous solution were determined through an UV/vis spectroscopic study employing the SQUAD program as a computational tool. A NMR study complements the pK a values assignment and evidences a H- bridge presence on 1-C. The chemical model used was consistent with the experimental data obtained. The pK a values determined with this procedure were as follows: H 4 (MGF) = H 3 (MGF) - +H + ,pK a 1 (6-H) = 6.52 ± 0.06; H 3 (MGF) - =H 2 (MGF) 2- +H + ,pK a 2 (3-H) = 7.97 ± 0.06; H 2 (MGF) 2- = H(MGF) 3- +H + ,pK a 3 (7-H) = 9.44 ± 0.04; H(MGF) 3- = (MGF) 4- +H + ,pK a 4 (1-H) = 12.10 ± 0.01; where it has been considered mangiferin C 19 H 18 O 11 as H 4 (MGF). Mangiferin UV/vis spectral behavior, stability study in aqueous solution as well as NMR spectroscopy studies: one-dimensional 1 H, 13 C, 2D correlated 1 H/ 13 C performed by (g)-HSQC and (g)-HMBC methods; are also presented. pK a values determination of H 4 (MGF) in aqueous solution is a necessary contribution to subsequent pharmacokinetic study, and a step towards the understanding of its biological effects. © 2005 Elsevier B.V. All rights reserved. Keywords: Mangiferin; Acidity constants; UV/vis spectroscopic determination; 1 H NMR; 13 C NMR 1. Introduction Mangiferin (1,3,6,7-tetrahydroxyxanthone-C 2 --D-glucosi- de) (Scheme 1) is a glucoside xanthone widely distributed in higher plants in families such as Anarcardiaceae [1], Gen- tianaceae [2] and Guttiferae [3]. Some of these have been rec- ommended as traditional medicinal plants [4,5]. Recently, in Cuba, it has been developed an aqueous decoction of mango (Mangifera indica L.) stem bark used as a nutritional supplement and phytomedicine registered as Vimang ® [7]. The Cuban stud- ies underline its anti-tumoral effect. Something similar occurs with Salaretin ® , a Sri Lanka’s product that can be used in the obesity treatment and particularly on diabetes type II [8,9]. Actu- ally, in the present, the impact about the mangiferin is due to its pharmacological effects as an anti-tumoral, anti-viral, and anti- Corresponding author. Tel.: +52 55 58044670; fax: +52 55 58044666. E-mail address: suemi918@xanum.uam.mx (A. Rojas-Hern´ andez). diabetic agent [6,10–16]. The mangiferin bioactivities observed have been related to some antioxidative as well as free radi- cal captodative functions [17–19] since its structure seems to be associated with certain flavonoid systems and their activities [20,21]. However, the studies about mangiferin generally are done from the pharmacological perspective, not from the physico- chemical point of view. The previous UV/vis and NMR studies were focused on its identification [22–25]. There is no infor- mation about the mangiferin equilibrium constants, pK a values or complexation constants with inorganic cations. The present work is focused on obtaining at least some of this informa- tion. In this study, we considered some graphic methods and the use of computational programs to determine the equilibrium constants. SQUAD [26] program has been used before by our research group, and excellent results for UV/vis spectroscopic data [27–29] have been obtained. Thus, a similar approach was undertaken for the equilibrium constants of mangiferin in aque- ous solution. pK a values determination of H 4 (MGF) in aqueous 1386-1425/$ – see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.saa.2005.09.009