Anthocyanins in red cabbage -
The optimum extraction efficiency was obtained at ultrasonic output power of W, pulse mode of 30 s ON: 30 s OFF, at 15 °C for 90 min. Fractionation of crude extract was further carried out using three different concentration of ethanol in ion-exchange column chromatography HP This is a preview of subscription content, log in via an institution to check access.
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Manley, S. Comunian, R. Ravanfar, I. de Castro, R. Dando, C. Favaro-Trindade, A. Selig, A. Dando, A. Alcaine, A. Shirsath, S. Sonawane, P.
Gogate, Chem. Chandrasekhar, M. Madhusudhan, K. Raghavarao, Food Bioprod. Xavier, T. Lopes, M. Quadri, M. Quadri, Braz. Vinson, B. Hontz, J. Food Chem. Agbor, J. In batch procedures, agitation improves the mass transfer and the time needed to achieve a dye concentration that provides an economical viable process Cacace and Mazza, The column process is a dynamic batch process where the solid phase raw material is stationary in a fixed porous bed, and the liquid phase solvent moves through the bed by forced convection.
In column extraction, the solvent can be recirculated at different flow rates or operated in one single step.
In both the situations, a concentration gradient is established throughout the column due to the contact of the solvent with the raw material. Extraction occurs until the dye concentration in the solvent reaches equilibrium with the raw material.
Column extraction in one single step is completed when all colorant is removed from the raw material. The objective of this work was to investigate the variables influencing the column extraction of red cabbage anthocyanins aiming to determine the optimum operation conditions.
Red cabbage, a widely cultivated vegetable in all the regions of Santa Catarina, south of Brazil, was obtained from the local market. Batch assays were carried out macerating chopped red cabbage in ml of different solvents in a shaker for 24h, followed by the filtration.
The filtrate was centrifuged at rpm for 15 min. To evaluate the effect of pH on the bathochromic and hyperchromic behavior of the anthocyanins, 12 values of pH, from 2.
The wavelength and the absorbance intensity at the maximum peak of absorption were analyzed in order to obtain a stable absorbance range.
Once defined the best proportion of solid and liquid in the static assays considering maximum extraction at a minimal cost, the concentration of acetic acid was evaluated.
Experiments were carried out in triplicate and the dye concentration was expressed as the equivalent concentration of Congo red mg L -1 , as proposed by Sondheimer and Kertesz Column tests were performed with or without recirculation of the solvent.
The column extraction system with no solvent recirculation was comprised of a 20 L tank where a submerged pump pushed the solvent through a cylindrical glass column in an ascendant flow. A good flow distribution at the entrance of the column of 3 cm internal diameter × 30 cm height was guaranteed by a perforated glass plate and a layer of 2 cm height made of glass beads of 4 mm diameter, as shown in Figure 1.
The porous bed of 25 cm height was formed of chopped red cabbage particles with the same dimensions used for the static essays, mixed with the glass beads at a mass ratio of g g The column ontained 70 g maximum quantity of 7. At the top of the column, a 2 cm layer of glass beads was also added to minimize dragging of the cabbage particles.
A flow control device was placed after the column and 5 mL samples of the eluted solutions were collected over 48 hours to determine the dye concentration. Exhaustion assays with no recirculation of red cabbage anthocyanin with a pre-determined solvent were carried out at different flow rates: 0.
Only the first flow rate showed to be economically viable due to the very high consumption of acetic acid at the two other conditions. In the extraction system using recirculation, a reservoir of 2 L was used instead of the 20 L tank, as in the previous arrangement.
Samples of 1 mL were taken directly from the reservoir for the analysis. A factorial experimental design was performed to investigate the effects of the five factors shown in Table 1. A five level experimental design was used to optimize the significant factors selected from the first design Table 2.
It was observed that the increase in pH was followed by a l máx displacement in the visible range from pH 3. The intensity of the peak diminished, as observed by the reduction in the absorbance. According to Brouillard , the largest coloring changes of the anthocyanins occur near to the pK values where the concentration of the flavilium cation and the quinoidal base are equal.
The flavilium cation is red, while the quinoidal base is blue, and the pH variations modify the equilibrium of the proton transfer between these two chromophores. At pH 2. The results of the Duncan test were obtained with the aid of Statistica ® Software. No bathochromic shift was observed from pH 2.
The above two factors must be associated to give results within a steady range of readings, aiming to produce no bathochromic effect, and obtain, at the same time, hyperchromic results that fit the Beer law. A convenient value was observed at pH 3. At this value, which was also recommended by Sondheimer and Kertesz and Lopes , a high absorbance was obtained.
Cabrita et al. A gradual bathochromic shift from to nm was found when pH varied from 1. The extraction capacities of the several solvents studied are shown in Table 4.
Since it was the most effective solvent, it was chosen to be used in further tests. Vegetable tissue is comprised of cells joined by middle lamellae, compounds of which are associated with celluloses, hemicelluloses, lignins and pectic substances.
Treatments with acid solutions break the hydrogen bonds between these compounds, destroying the selective permeability of the red cabbage cell, and the solutes easily leave the cell. As acetic concentration increases, sugar hydrolysis also rises, and more anthocyanin is extracted. It appeared that the acetic acid was more toxic to the cell than the McIlvaine buffer since the 5.
These results explained the lower extraction rate found in this study, at high ethanol concentrations Table 4. The results of Duncan test for the solvents are also given in Table 4 , at superscript letters, where the comparison for the different solvents and the resulting extracted colorant concentration are shown.
It gave a significant difference from all other solvents. The best pH for the extraction considering the mass extracted has not been well established in the literature, but extraction at low pH values has always been recommended.
Using ethanol-water mixtures, the most common applied solutions for extraction, there was no significant difference between the concentrations used. The extraction capacity was similar to the buffer solutions at pH 4. Since an increase in the cabbage mass occured in the mixture used for the extraction, the anthocyanins content in the extract also increased, but it had no linear response.
The best combination was found in the range of 0. In this interval, dye concentration decreased from 1 to 0.
Changes in the solvent:solid ratio alter the final equilibrium between the dye concentrations in the solid and liquid phases. The performance of the process is also affected by the extraction conditions, the solubility modifications and the solute:solvent interactions Cacace and Mazza, As a consequence, the principal effect of the solvent:solid ratio is to modify the equilibrium constant, and thus to improve the anthocyanin extraction when a greater solvent:solid ratio is used Table 5.
The influence of acetic acid on dye extraction is shown in Table 6. A non linear relationship between the increase of extracted mass and acid concentrations was indicated by the results. The significant differences obtained under the studied conditions are shown by results of Duncan test, indicated by the superscript letters.
The mass of colorant was exhausted from the red cabbage using tests without recirculation of the solvent, and 1. Figure 2 shows the mass evolution of dye extracted from red cabbage and the colorant concentration at the outlet of the column operating at 0.
Complete extraction needed more than 33 h of the processing. In order to study the effect of the five variables chosen, salt concentration SC NaCl [g L -1 ], pH, flow rate, FR [L h -1 ], solvent volume, SV [L], and mass of red cabbage, CM [g], a factorial experimental design was carried out, and the anthocyanin concentration Conc.
The analysis of the effects indicated that the pH, the volume of solvent and the mass of red cabbage were significant boldface since the p-level was lower than 0.
Table 8 also shows the t-test and the confidence limits. From these results, a central composite design was proposed in order to optimize the anthocyanin extraction in the column with a recirculation system. Sixteen tests Table 9 were run at 6 L h -1 recirculation flow rate to obtain a response surface.
The values found for the effects showed that: a pH and SV had negative effects on the concentration of the extracted dye, b CM had a positive one, and c SC and FR were inert variables.
The negative effects showed that as the pH increased lower dye mass was extracted, and as the solvent volume indreased from 1. On the other hand, an increase of the mass of red cabbage from 30 to 50 g gave a significant increase in the dye concentration because there was more dye source available for extraction.
FR was set on 6. Sodium chloride SC was not used hereafter. Considering the significant terms only, the fitted reduced model Equation 1 had a determination coefficient R 2 of 0. The residual square sum, SSr, was smaller compared to that of the regression, SSR, indicating that the model was suitable, which explained Residual values were not significant, and the model could be used for prediction Montgomery, of dye concentration extraction in similar situations.
The contour graphs generated by the model are shown in Figure 3 for the pH and SV as a function of the red cabbage mass, CM.
The optimal condition range was observed in the Figure 3. It varied from 40 to 50 g for CM, from 2. In this studied range, the optimal conditions were pH 2. At the exhausting assay, the concentration was 2. The major advantage of using the solvent recirculation system was that the higher the concentration of the dye in the liquid phase, the lower the energy consumption needed to concentrate and purify the pigment.
Although anthocyanin extraction is a subject that is of concern to the scientific community, processes employing static methods are generally used. Almost no information can be found for the extraction in columns. This paper proposes that anthocyanin extraction from red cabbage in columns is a viable process, inexpensive and relatively quick, providing a final extracted dye solution of such a high concentrated that it can be commercialized immediately.
Also, the solvent applied is the diluted acetic acid that is largely used as vinegar.
Ancient healing therapies pigments, called anthocyanins, provide fruit and vegetables Anfhocyanins Benefits of vitamin D blue, purple and Ajthocyanins coloring, and Anthocyanins in red cabbage a group of healthful compounds that fall within Protein intake and hunger control flavonoid class of Antjocyanins nutrients. ARS scientists erd identified 36 Muscle definition diet in red cabbage, including eight that have never before been detected in the cabbage. The study was conducted at the ARS Beltsville Human Nutrition Research Center BHNRC in Beltsville, Md. According to the scientists, emerging evidence suggests that anthocyanins may provide cancer protection, improve brain function and promote heart health. An earlier ARS study showed that some anthocyanins yield twice the antioxidant power of the same amount of vitamin C in test tubes, although the amount absorbed by the human body was not explored.
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