Among them, carotenoids such as for example halocynthiaxanthin and fucoxanthinol of are reported to exhibit antiproliferative activity against human leukemia, breast and colon cancer cells (Konishi et al. VBPintestine and VBPblood plasma against \glucosidase were 28.34 and 12.60?g/ml, respectively. The values of VBPintestine and VBPblood plasma were 4.29, 0.036, 6.58 and 1.53??103, and 7.63?mM, 0.057?mM/min, 10.41?s?1 and 1.36??103 (M?sec)?1, respectively. There Asiaticoside was a synergistic interaction between VBPblood plasma and VBPintestine on rat intestinal -glucosidase inhibitory activity. Electronic supplementary material The online version of this article (10.1007/s13197-018-3099-9) contains supplementary material, which is available to authorized users. recorded the highest concentration of 350?mM which was 107 times higher than that of sea water (Ueki and Michibata 2011). The vanadium concentrations in the internal tissues and blood of the solitary ascidian were higher than that of the whole tunic, and at low pH values, vanadium metabolites exerted chemical defense in the ascidian Rabbit Polyclonal to mGluR4 (Odate and Pawlik 2007). Several proteins are known to be involved in the accumulation of vanadium in ascidians (Yoshinaga et al. 2007). Among them, vanabin family in blood cells (vanadocytes) and coelomic fluid of the ascidian gained much attention due to their selective binding ability for vanadium (Ueki et al. 2003, 2008). A vanadium binding protein of the sea cucumber intestine had adipocyte differentiation inhibitory effect which was mediated by activating the WNT/-catenin pathway (Liu et al. 2015a). Inorganic vanadium (IV) and (V) complexes and synthetic vanadyl complexes with organic ligands are known to have antioxidant activities (Francik et al. 2011; Wazalwar and Bhave 2012). But the antioxidant activities of vanadium binding proteins have not been investigated so far. The sea squirt is a popular sea food in Korea and Japan, in which many bioactive Asiaticoside compounds have been extracted. Among them, carotenoids such as halocynthiaxanthin and fucoxanthinol of are reported to exhibit antiproliferative activity against human leukemia, breast and colon cancer cells (Konishi et al. 2006). However, there is no comprehensive study on vanadium binding proteins of the sea squirt so far. Therefore, the objectives of this study were to determine the vanadium concentrations in the tissues of and \glucosidase and rat intestinal acetone powder were purchased from Sigma Chemical Co. (St. Louis, MO, USA). DEAE Sepharose fast flow and Sephacryl S-200 HR gels were purchased from GE Healthcare (Uppsala, Sweden). All other chemicals used in this study were of analytical grade. Sample preparation Fresh sea squirt obtained from a local fishery market (Gangneung, Korea) packed on ice was transported to the laboratory within 1?h. Blood plasma, intestine and muscle tissue of the sea squirt were separated, immediately deep frozen (at ??80?C) and stored at ??25?C until use. Determination of vanadium concentration Samples (0.2?g) in a Kjeldahl flask were mixed with 10?ml of water and 2?ml of nitric acid. This mixture was gently heated for 3C5?min. After cooling, 5?ml of sulfuric acid was added and heated again until the yellow color became colorless. After cooling down to room temperature, 30?ml of water Asiaticoside and 10?ml of saturated ammonium hydroxide solution were added, and then heated until the white smoke occurred. Ten ml of the decomposed solution was mixed with 2?ml of 25% ammonium citrate and 2?ml of bromothymol blue, neutralized with aqueous ammonia, and then mixed with 2?ml of 40% ammonium sulfate. Afterwards, 1?ml of 10% diethyl dithocarbamic acid was added and mixed well. After adding 20?ml of chloroform, it was vigorously shaken and then the chloroform layer was separated. This was repeated with 20?ml of.