Betalains Assay of Fermented Red Beet Root Extract by High Performance Liquid Chromatography

Abstract

A high performance liquid chromatographic process was developed for betacyanin and betaxanthin pigments in a fermented red beet root extract. The method employs a reverse-phase microparticular C& column, ion pairing, and a methanol-water mobile phase, using a solvent gradient program. Spectrophotometric evidence and an enzymatic method were used to unequivocably identity peaks obtained from LC chromatograms. This accurate method for betalains separation and quantification was rapid and efficient. This method of assay enables the stability of the colorant in drug forms to be monitored routinely.

Aqueous two phase extraction for purification and concentration of betalains

“…For the HPLC chromatograms of betalains detected at 538 nm, the peaks are (1) betanin, (2) isobetanin, (3) betanidin, (4) isobetanidin, (5) prebetanin and for the HPLC chromatograms of betalains detected at 478 nm, the peaks are (1) vulgaxanthine I, (2) vulgaxanthine II, (3) betanin, (4) isobetanin, (5) betanidin, (6) isobetanidin and (7) prebetanin. The chromatograms obtained showed the peaks are similar to that of the fermented beet juice (Pourrat et al, 1988). The HPLC chromotograms depicts that betalains are concentrating in the top phase of the aqueous two-phase system.…”

“…The individual components of the betalains after the extraction, was determined by HPLC method (Pourrat, Lejeune, Grand, & Pourrat, 1988). C-18 ODS column (250 mm  4.6 mm SGE, Japan,) having a pore size of 5 lm was used for HPLC analysis of betalains.…”

Betalains and Phenolics in Red Beetroot (Beta vulgaris) Peel Extracts: Extraction and Characterisation

“…O ther betacyanins reported in red beetroot m aterials are betanidin, isobetanidin, isobetanin, prebetanin, neobetanin, amaranthin, lam pranthin I and lampranthin II (Alard et al, 1985;Pourrat et al, 1988;Bokern etal., 1991;Jackman and Smith, 1996). The solvents used to extract betalains are mainly water and aqueous methanol (50-80% ).…”

Inactivation of peroxidase and polyphenol oxidase in red beet (Beta vulgaris L.) extract with high pressure carbon dioxide

“…The HPCD treatment resulted in the contents of betanin and isobetanin, the two compounds mainly responsible for the red color of red beet (Pourrat et al, 1988), to decrease by 7.13% and 12.79%, respectively, which were lower than the reductions caused by blanching. The HPCD treatment also resulted in a loss of approximately 17% and 19% of vulgaxanthin I and II, respectively, which are mainly responsible for the yellow hue of the RBE (Pourrat et al, 1988). In contrast, the betaxanthin content in the blanched RBE samples increased by about 14%, suggesting that other compounds might be converted to betaxanthins.…”

“…The absorbance of the samples at 480 and 535 nm was measured to calculate the color shade value (ratio of absorbance at 480 to that at 535 nm (Stintzing, Schieber, & Carle, 2003). Betanin, isobetanin, vulgaxanthin I and vulgaxanthin II in the samples were determined by high performance liquid chromatography-mass spectrophotometry (HPLC-MS) following the method of Pourrat, Lejeune, Grand, and Pourrat (1988) and Herbach et al (2004) with minor modifications. HPLC-MS was performed with an Agilent 1100 HPLC-ESI-MS system.…”