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1

Study of Optical and Electrochemical Properties of Solvent-Dependent Natural Dye Extracted from Rivina humilis L

Aliah Hasniah, Iman Ryan N., Sriwidati Sriwidati, Sawitri Asti, Setiawan Andhy, Putri Assa P. D., Kurniawati Fera

Journal of Ecological Engineering

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2023

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Vol. 24, nr 9

312--321

EN

This work aimed to study the natural dye extracted from Indonesian wild plants (Rivina humilis L.) using different solvents. The natural dye was extracted using the maceration method. Three different solvents, namely, aquades, acetone, and ethanol 96%, were used to extract natural dye from Rivina humilis L fruit. The absorbance spectra of the extracted dye were recorded using Ultraviolet-Visible (UV-Vis) spectroscopy. The different spectra of betalain pigment revealed the dye extract’s dependence on the solvent. The functional groups of the extracted dye were analyzed using Fourier transform infrared (FTIR) spectroscopy. The adherence of carbonyl and hydroxyl groups from FTIR spectra indicated that this dye could anchor to a semiconducting material, e.g., TiO2, which was commonly used in dye-sensitized solar cells (DSSC). The electrochemical properties of the extracted pigments were studied through higher occupied molecular orbital (HOMO) and lower unoccupied molecular orbital (LUMO) energy levels. Based on the results, the best performance to construct DSSC was achieved by natural dye adsorption with aquades solvent.

2

Przeciwprądowa chromatografia cieczowa w rozdzielaniu betalain

Spórna-Kucab A., Wybraniec S.

Przemysł Chemiczny

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2016

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T. 95, nr 10

1904--1906

PL

Dokonano przeglądu literaturowego zastosowań przeciwprądowej chromatografii cieczowej w rozdzielaniu betalain (barwniki naturalne). Dodatkowo zwrócono uwagę na cenne właściwości prozdrowotne betalain i innych związków naturalnych.

EN

Areview, with 36 refs. Some therapeutic properties of betalains were also taken into consideration.

3

Separation of betalains by gradient high-speed counter-current chromatography

Spórna-Kucab A., Udziela D., Sobocińska M., Szot D., Wybraniec S.

Challenges of Modern Technology

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2016

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Vol. 7, no. 1

23--28

EN

A study on separation of betalain mixture obtained from red beet juice (Beta vulgaris L.) by analytical high-speed counter-current chromatography (HSCCC) was performed. The extract was obtained by thermal treatment of acidified red beet juice for 30 min in 85 °C. The pigment mixture consisted of betanin/isobetanin as well as their decarboxy- and dehydro-derivatives. The HSCCC process was accomplished in the ‘tail to head’ mode with two polar solvent systems containing salt: BuOH-EtOH-NaClsolution-H2OH3PO4 (1300:700-1000:1300:700:2.5-5.5 (system I), 1300:200-400:1300:700:2.5-4.5 (system II); v/v/v/v/v). The retention of the stationary phase was 73% (system I) and 79% (system II). The mobile phase was pumped at 2 ml/min flow rate. HPLC-DAD-ESI-MS analyses were performed in reversed phase mode for the obtained HSCCC fractions and crude extract. The solvent systems enabled separation of betanin and decarboxy-betanins (system I and II) as well as neobetanin (system II). Additionally, some pure fractions of 17-decarboxy-betanin and 2,17-bidecarboxy-betanin were obtained in system II.

4

Research on solvent systems with tetra-n-butylammonium bromide for counter-current chromatography of betalains

Spórna-Kucab A., Wysoczańska J., Bartel A., Świergosz T., Szot D., Wybraniec S.

Challenges of Modern Technology

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2016

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Vol. 7, no. 1

12--16

EN

In this study, new two-phase solvent systems for counter-current chromatography (CCC) consisting of n-butanol and water as well as various amounts of acetic acid, acetonitrile, ethanol, acetone or ethyl acetate were tested. Additionally, tetra-n-butylammonium bromide (TBAB) was introduced into the system in the form of aqueous solutions or phosphate-citrate buffer (pH 6.7) in order to form ion-pairs with betalains. The selection of buffer pH was based on their ability to create ion pairs by tetraalkylammonium salts, with selected betalains under these conditions. In this study, it is shown that the settling time of two phases is longer with the increase of acetic acid/acetonitrile/ethanol/acetone/ethyl acetate. For selected solvent systems with high amounts of acetonitrile, ethanol and acetone two phases were not observed. The systems with acetone have the largest increase of settling time. Ethyl acetate systems were characterized by a slow settling time increase. In systems containing additionally 2% aqueous TBAB, smaller changes in settling time than in similar systems without TBAB were observed. Addition of TBAB in the buffer resulted in a prolongation of settling time. Solvent systems in which the separation between the aqueous and organic phases was visually best, were selected from among all the tested systems and the betalain partition coefficients were measured by LC-DAD-ESI-MS. The best results were observed for systems: n-butanol-water-acetic acid (2:2.5:0.75, v/v/v), n-butanol-water-acetic acid (2:2.5:1, v/v/v) and n-butanol-TBAB in water-acetonitrile (2:2.5:0.5, v/v/v).

5

Enzymatic oxidation of neobetanin monitored by liquid chromatography with mass spectrometric detection

Starzak K., Szot D., Skopińska A., Swiergosz T., Wybraniec S.

Challenges of Modern Technology

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2016

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Vol. 7, no. 1

29--31

EN

The aim of this study was monitoring of enzymatic oxidation of neobetanin, an interesting type of betalains which is a partially oxidized betacyanin. As it belongs to betalains, it is water soluble and non-toxic, but a presence of a few functional groups makes it very reactive. Oxidation reactions were performed using horseradish peroxidase followed by spectrophotometric and mass spectrometric detection (LC-DAD-ESI-MS/MS) of obtained products. Enzymatic oxidation of neobetanin leads to a formation of new decarboxy- and dehydro-derivatives. The main identified oxidation product is 2-decarboxy-2,3-dehydroneobetanin. Searching for all formed oxidation products is extremely important for elucidation of the betalains oxidation mechanism.

6

The effect of ascorbic acid supplementation on betacyanin stability in purple pitaya (Hylocereus polyrhizus) juice

Skopińska A., Szot D., Starzak K., Mizrahi Y., Wybraniec S.

Challenges of Modern Technology

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2015

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Vol. 6, no. 4

14--18

EN

Current studies on betalains are focused on searching of new plant sources of these pigments. In this light, purple pitaya (Hylocereus polyrhizus) is a valuable batalain-containing fruit. Betalains are known to be sensitive at elevated temperatures conditions. Nevertheless, addition of specific food stabilizers (EDTA, ascorbic acid, citric acid) as well as natural matrix compounds may exert stabilizing effect on maintenance of pigments. Therefore, in presented study, the stability of betalains in Hylocereus polyrhizus juices with protective addition of ascorbic acid at 85 ̊C was examined. Spectrophotometric analyses enable monitoring of reaction during heating of solutions. The hypsochromic shift of λmax to 460 nm is observed in more acidic media without ascorbic acid, suggesting the generation of new reaction products. These absorption bands in solutions with ascorbic acid are not detected, indicating on retaining of the basic structure of betalain chromophore. Generally, pigment retention is diminishing with prolonged heating and depends on pH-values of tested solutions. The addition of ascorbic acid significantly affects the betalain stability. After 60 min of heating, more than 60% of the initial pigment is retained at pH 3-4 in samples of purple pitaya with ascorbic acid. In summary, betacyanins in purple pitaya juice are shown to exhibit proper heat stability, when are stabilized with ascorbic acid. However, a protective effect of natural juice matrix and ascorbic acid is more significant at acidic pH.

7

Semi-synthesis of red beet betacyanin ethyl-esters by esterification

Tuwalska D., Starzak K., Szot D., Wybraniec S., Winterhalter P., Jerz G.

Challenges of Modern Technology

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2014

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Vol. 5, no. 3

27--31

EN

Red beet pigments recovered from Beta vulgaris L. (Chenopodiaceae), mainly consist of purple betacyanins, such as 15R/15S-betanin. Currently,this pigment class is intensively investigated in respect of stability in food systems. Quite popular is the use of red beet extracts in dairy products such as yoghurts, and ice-creams. So far, there are no issues of toxicity using these natural pigments in food – so they seem to be a very good alternative to replace synthetic dyes which had been frequently discussed for negative side-effects to human health. Betacyanins, the glycosidation products of betanidin are very polar and therefore water soluble pigments, which are biosynthetically derived from condensation of cyclo-DOPA and betalamic acid. The significant polarity of betacyanins requires the use of ion-pair reagents such as trifluoro-acetic acid (TFA) or other homologue fluorinated additives under standard separation conditions on larger scale in spiral coil countercurrent chromatography (spCCC). In our synthetic experiment, the polarity of the natural betacyanins was reduced. This omitted the use of toxic per-fluoro ion-pair reagents. The three carboxylic groups of the betanin pigment backbone were the target functional groups of the derivatisation. In semi-synthesis, fortified red beet pigment extract was esterified with water-free ethanolic hydrogen chloride solution. The reaction was carried out at ambient temperature (7 days) to yield a mixture of betacyanin mono-, di- and tri-ethyl-esters, and their epimeric forms, as well as the respective betanidin-ethyl-esters. The time course of the reaction mixture was monitored by analytical C18-HPLC with ESI-DAD-MS/MS detection. The detected molecular weights of the pigments confirmed the presence of expected products. To the best of our knowledge, the presence of betalains containing ethyl-ester groups in biological samples have not been studied before.

8

Mass spectrometric detection of products of decarboxy-betanins UV-irradiation

Starzak K., Skopińska A., Szot D., Wybraniec S.

Challenges of Modern Technology

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2014

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Vol. 5, no. 3

22--26

EN

Betalains are a group of natural plant pigments, responsible for red-violet colour of numerous flowers, fruits and roots. They commonly occur in order of Caryophyllales (Caryophyllales). Recent studies revealed great health promoting properties of these pigments, but their natural origin make them very labile under various physicochemical conditions. In the case of betanin degradation, many new, decarboxy-betanins are formed. Latest study showed that UV-irradiation had significant impact on betanin stability, however, no deeper studies were performed in this field. In order to know an impact of the process conditions on degradation of betanin and its decarboxylated derivatives, series of experiments with UV-photo-irradiation was performed in aqueous solutions as well as in various organic solvents at wide range of pH. The highest pigment stability was observed in aqueous solutions for betanin and 17-decarboxybetanin at pH 4.5–7, and for 2,17-didecarboxybetanin at low pH. In all investigated organic solutions, all of the studied compounds have been degraded. The LC-DAD-ESI-MS analysis identified new reaction products. The results showed that UV-photo-irradiation leads mainly to a formation of many new and various 14,15-dehydrobetanins.

9

Wpływ warunków ogrzewania roztworów koncentratu soku z buraka ćwikłowego na parametry barwy i zawartość barwników betalainowych

Gościnna K., Walkowiak-Tomczak D., Czapski J.

Aparatura Badawcza i Dydaktyczna

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2014

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T. 19, nr 2

183-189

PL

Celem pracy było określenie wpływu ogrzewania roztworów koncentratu soku z buraka ćwikłowego rozcieńczonego do zawartości ekstraktu 14%, na zmiany parametrów barwy i barwników betalainowych. Zmiany te badano w zależności od pH roztworu, czasu i temperatury ogrzewania. Badania przeprowadzono z zastosowaniem metody płaszczyzn odpowiedzi wykorzystując układ doświadczalny Box-Behnkena. W czasie ogrzewania następowała degradacja fioletowych barwników betalainowych, tym większa, im dłuższy był czas i temperatura ogrzewania. Na zawartość żółtych barwników betalainowych największy wpływ miało pH. Największą stabilność barwników betalainowych stwierdzono w próbach o pH 4,5. Wartości składowych barwy X, Y, Z wzrastały w czasie ogrzewania, co związane jest ze spadkiem zawartości barwników fioletowych. Parametr L* określający jasność, a* określający udział barwy czerwonej oraz parametr b* określający udział barwy żółtej wzrastały w czasie ogrzewania. Największy wpływ na zmianę tych wyróżników miał czas ogrzewania, a najmniejszy wpływ miało pH. Wartość tonu barwy h* wzrastała w czasie ogrzewania, co wskazuje na zmianę barwy w kierunku żółtej.

EN

The aim of this study was to determine the changes of colour parameters and betalain pigments contents in solutions of red beet juice concentrate (14% extract) during heating in dependence on pH, time and temperature of heating. Analyses were conducted in model systems using the response surface method according to Box-Behnken design. During heating of solutions, violet betalain contents decreased with an increase in time and temperature of heating. The pH value had the greatest effect on yellow betalain pigment contents. The highest stability of betalains was determined in pH 4,5. Values of the X, Y and Z components increased during heating, what was associated with decreasing of violet pigment contents. Lightness of colour L*, the share of the red colour a* and share of the yellow colour b* increased during heating. The time of heating had the greatest effect on these responses, the effect of pH value was relatively small. Values of hue angle h* increased, what indicate a change in colour tone towards yellow.

10

Stability of Betanidin Influenced by Copper Cations

Tuwalska D., Starzak K., Szot D., Skopińska A., Wybraniec S.

Logistyka

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2013

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nr 4

EN

Latest studies revealed some antioxidant properties of betalain pigments that contribute to the growth of interest in these compounds. During the isolation and storage, betalains are exposed to many different chemical and physical degradative factors, which often lead to their decomposition and discoloration. An effect of copper cations on betanidin stability was investigated in aqueous and organic-aqueous solutions. Betanidin stability experiments were performed at different pH’s, with different concentrations of metal cations and in different solvents. Using the modern spectrophotometric microplate reader it was possible to monitor changes of absorption spectra of the pigment samples during their incubation with copper ions in parallel at various physico-chemical conditions. Incubation with copper cations leads to betanidin degradation in all investigated solvents in the whole range of pH. The formed products are its new oxidized derivatives. A possibility of a formation of metal-pigment complexes is also considered. Recognition of betalains degradation paths is important because of their not yet fully recognized antioxidant and chemopreventive properties.

PL

Ostatnie badania wykazały przeciwutleniające właściwości betalain, co przyczyniło się do wzrostu zainteresowania tą grupą związków. Podczas izolacji i przechowywania, betalainy są narażone na wiele różnych degradujących czynników, chemicznych i fizycznych, które często powodują ich rozkład i odbarwienie. Zbadano wpływ kationów miedzi na stabilność betanidyny w roztworach wodnych i wodno-organicznych. Eksperymenty były przeprowadzane w różnym pH, z różnymi stężeniami kationów metalu i rozpuszczalników organicznych. Zastosowanie nowoczesnego spektrofotometrycznego czytnika mikropłytek pozwoliło na monitorowanie zmian w widmie absorpcyjnym pigmentu w różnych warunkach fizyko-chemicznych. Inkubacja betanidyny z jonami miedzi prowadzi do jej degradacji we wszystkich badanych rozpuszczalnikach w całym zakresie pH. Powstałe produkty są jej nowymi, utlenionymi pochodnymi. Rozważano również powstawanie kompleksu metal-pigment.Poznanie szlaków utleniania betalain jest ważne z powodu nie w pełni jeszcze poznanych ich antyutleniających i chemoprewencyjnych właściwości.

11

Spiral-coil countercurrent chromatographic separation of betanin and betanidin ethyl-esters monitored by continuous ESI-MS/MS injection and LC-ESI-MS/MS

Tuwalska D., Szot D., Wybraniec S., Winterhalter P., Jerz G.

Challenges of Modern Technology

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2013

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Vol. 4, no. 4

21--25

EN

Betalains present in Beta vulgaris L. (Chenopodiaceae) have a high potential as natural pigments for food applications. They have been used as a substitute of synthetic dyes in processing of gelatine, strawberry yogurts, ice creams, fruits cocktails, candies and biscuits. The stability of betalains is strongly influenced by numerous factors – sugar content, and impact of light, oxygen, water activity, pH and temperature. A mixture of betacyanin mono-, di- and tri-ethyl-esters was separated by a large-volume spiral-coil countercurrent chromatography (spCCC) prototype which yielded fractions continuously off-line injected to anÂ ESI-MS/MS device. This method yielded a reconstituted pigment profile and finally enabled the MS-target-guided isolation procedure for the whole spCCC-experiment. The individual fractions were analyzed by LC/ESI-MS/MS. Betanin and betanidin mono-, di- and tri-ethyl-esters as well as other decarboxylated derivatives were detected in the chromatograms. The ethyl-esters of betanin were only partially fractionated, but clearly fractionated from the ethyl-esters of betanidin which will be of great value for biological evaluations.

12

Zmiany aktywności przeciwutleniającej, zawartości barwników betalainowych i polifenoli w roztworach koncentratu soku z buraka ćwikłowego podczas ogrzewania

Gościnna K., Czapski J., Walkowiak-Tomczak D.

Aparatura Badawcza i Dydaktyczna

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2013

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T. 18, nr 4

373-379

PL

Celem badań było określenie wpływu czasu, temperatury i pH roztworów koncentratu z soku buraka ćwikłowego rozcieńczonego do zawartości ekstraktu 14% na zmiany zawartości barwników betalainowych, polifenoli i aktywności przeciwutleniającej. Doświadczenie zaplanowano z zastosowaniem metody płaszczyzn odpowiedzi, w układzie doświadczalnym Box-Behnkena przy trzech poziomach zmiennych niezależnych: temperatury ogrzewania (70-90°C), czasu ogrzewania (0-120 min) i wartości pH (3-6). Największą stabilność barwników stwierdzono w próbach o pH 4,5. Istotny wpływ na zmianę zawartości barwników, na podstawie współczynników równań płaszczyzn odpowiedzi, miała temperatura i czas ogrzewania oraz interakcje między nimi. Temperatura i czas ogrzewania nie miały istotnego wpływu na zawartość polifenoli. Istotny był natomiast wpływ pH. Zaobserwowano wysoką korelację pomiędzy zawartością barwników betalainowych, a zdolnością przeciwutleniającą badanych prób. Największy wpływ na zdolność przeciwutleniającą miała temperatura ogrzewania. Dla większości zmiennych zależnych uzyskano dobre dopasowanie dla płaszczyzn drugiego stopnia. Metoda płaszczyzn odpowiedzi w badanym zakresie czynników doświadczalnych umożliwia dobre przewidywanie zmian zawartości związków biologicznie aktywnych oraz zdolności przeciwutleniającej w roztworach koncentratu soku z buraka ćwikłowego, w zależności od warunków ogrzewania.

EN

The aim of this study was to determine the effect of temperature, time and pH values on changes in betalain and polyphenol contents and antioxidant capacity in solutions (14% extract) of red beet juice concentrate during heating. Analyses were conducted in model systems with the use of response surface methodology, according to the Box-Behnken design for three independent variables: temperature (70-90°C), time (0-120 min) and pH (3-6). The highest stability of betalain pigments was determined in pH 4,5. The time and temperature of heating and interactions between them had the greatest effect on betalain pigments. pH value had the greatest effect on polyphenol contents, while temperature and time of heating had no significant effect on them. The temperature of heating had the strongest effect on antioxidant capacity. The pigment contents was significantly correlated with antioxidant capacity. The goodness of fit for quadratic equations was obtained for the most responses. Within the tested range of experimental factors the response surface method makes it possible to predict changes in bioactive compound contents and antioxidant activity, dependent on heating conditions.

13

Spectrophotometric investigation on betanin degradation induced by copper cations

Starzak K., Skopińska A., Wybraniec S.

Challenges of Modern Technology

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2012

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Vol. 3, no. 4

44--48

EN

Betanin pigment structurally belong to red-violet betacyanins which represent a class of N-heterocyclic water soluble plant pigments providing the colours in a wide variety of fl owers and fruits [1, 2]. Most of betacyanins, such as betanin, are 5-O-glucosides of betanidin (the basic chromophoric aglycone unit). Betanin colorant (E-162) is produced from red beet roots (Beta vulgaris L.) and is available in concentrates produced by evaporating beet juice under vacuum or as powders made by spray--drying the concentrate. In this study, an eff ect of copper cations on betanin stability was investigated in aqueous and organic-aqueous solutions. The presence of organic solvents (methanol, ethanol and acetonitrile) increased substantially the decomposition of the pigment induced by metal cations in acidic media, for which generally much lower degradation in aqueous solutions was observed. A possibility of a complex formation between betanin and Cu2+ was also stated at pH 7–8 and its relatively high stability in aqueous samples was observed. A presence of numerous products of betanin decomposition was noticed at the wavelength range of 380–500 nm in spectra obtained for the investigated metal cations, especially in the organic-aqueous solutions.

14

Research on betanidin oxidation by ABTS radicals

Starzak K., Skopińska A., Wybraniec S.

Challenges of Modern Technology

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2012

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Vol. 3, no. 4

39--43

EN

Betanidin is a basic betacyanin with 5,6-dihydroxyl moiety which causes its high antioxidant activity. It belongs to betalains, a group of natural, water-soluble plant pigments, which elicit a red-violet coloration of fruits and fl owers. One of the most popular sources of betanidin is red beet root (Beta vulgaris L.). Recent studies have shown importance of betalains oxidation, because of their high natural, antiradical and antioxidant activity and potential benefi ts for human health [1]. An eff ect of oxidation of betanidin using ABTS radicals was investigated in aqueous solutions at pH 3–8 and compared to activity of horseradish peroxidase. Products of the biomimetic betanidin oxidation were monitored by high performance liquid chromatography (HPLC) coupled to optical detection and mass spectrometry (LC-DAD-ESI-MS/MS). The presence of two main oxidation products: 2-decarboxy-2,3-dehydro-betanidin and 2,17-didecarboxy-2,3-dehydro- -betanidin at pH 3 indicates their generation through two possible reaction ways with two diff erent quinonoid intermediates: dopachrome derivative and quinone methide. Both reaction paths lead to the decarboxylative dehydrogenation of betanidin.

15

High salt-solvent systems in separation of betanin and its derivatives from red beet (Beta vulgaris L.) by high-performance countercurrent chromatography (HPCCC)

Spórna-Kucab A., Ignatova S, Garrard I., Wybraniec S.

Challenges of Modern Technology

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2012

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Vol. 3, no. 1

57--61

EN

A study on a separation of betanin and its decarboxy- and dehydro-derivatives obtained from red beet roots (Beta vulgaris L.) using analytical high-performance countercurrent chromatography (HPCCC — Dynamic Extractions Ltd., UK) was performed. The HPCCC process was accomplished in the ‘tail to head’ mode with three highly polar solvent systems with high salt concentrations: 1-propanol-acetonitrile-saturated ammonium sulphate-water (v/v/v/v, 1:0.5:1.2:1); ethanol-acetonitrile-1- propanol-saturated ammonium sulphate-water (v/v/v/v/v, 0.5:0.5:0.5:1.2:1) and ethanol-1-butanol-acetonitrile-saturated ammonium sulphate-water (volume ratio), 0.5:0.5:0.5:1.2:1). HPLC analysis was performed in a conventional reversed phase mode with diode-array (DAD) detection to characterize the composition of obtained fractions. The applied solvent systems enabled the separation of the betalain pigments with high efficiency for the first time. In the mode of separation selected, the more hydrophobic compounds eluted first as expected. Moreover, for the first time, the applied HPCCC solvent systems generated a separation of 2-decarboxy-betanin from 17- and 2,17-bidecarboxy-betanin as well as from neobetanin and betanin.

16

Zawartość barwników betalainowych, azotanów i zdolność przeciwutleniająca soków z buraka ćwikłowego w zależności od odmiany i wielkości korzenia

Gościnna K., Czapski J., Mikołajczyk-Bator K., Kidoń M.

Aparatura Badawcza i Dydaktyczna

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2012

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T. 17, nr 3

85-90

PL

Celem pracy była ocena zawartości barwników betalainowych, azotanów (III) i (V) oraz zdolności przeciwutleniającej soku z korzeni 14 odmian buraka ćwikłowego, o różnej masie korzeni. Surowiec podzielono pod względem masy na 4 grupy wielkościowe, a następnie przygotowano z nich sok. Stwierdzono, że zawartość barwników betalainowych oraz azotanów (V) w soku istotnie zależy od odmiany i wielkości korzenia buraka ćwikłowego. W badanych odmianach buraka ćwikłowego zaobserwowano duże zróżnicowanie zawartości barwników betalainowych i azotanów (V). Wraz ze wzrostem masy korzenia zawartość barwników betalainowych oraz zdolność przeciwutleniająca maleje. Zdolność przeciwutleniająca soku była istotnie skorelowana z zawartością barwników. Azotanów (III) nie wykryto w żadnej z badanych prób.

EN

The aim of this study was to estimate the content of betalain pigments, nitrate and antioxidant capacity of beetroot 14 cultivars, depending on the weight of root. The roots of red beet were divided into 4 groups according to their weight and then was prepared juice. It was found, that betalains and nitrate (V) contents of the juice significantly depends on cultivars and the size of beetroot roots. Large differences were found in betalains and nitrate (V) contents between cultivars. Betalains pigment contents and antioxidant capacity decreased with increasing root weight. The antioxidant capacity of juice is significantly correlated with pigment contents. Nitrate (III) were not detected in any tested samples.

17

Liquid chromatographic techniques in separation of betacyanins and their derivatives from red beet roots

Spórna A., Stalica P., Jerz G., Szaleniec M., Wybraniec S.

Challenges of Modern Technology

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2010

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Vol. 1, no. 1

19--22

EN

A study on separation of betacyanins and their decarboxylated as well as dehydrogenated derivatives obtained from red beet roots (Beta vulgaris L.) in high pressure liquid chromatography (HPLC) and ion-pair high-speed counter-current chromatography (IP-HSCCC) was performed. The IP-HSCCC process was accomplished in the ‘head-to-tail’ mode in a solvent system composed of butanol — acetonitrile — water (5:1:6 v/v/v, acidifi ed with 0.7% trifl uoroacetic acid). The HPLC separation was performed in a typical reversed phase mode with mass spectrometry (ESI-MS) and diode-array (DAD) detection. The chromatographic profi les of betalains obtained in these two techniques were signifi cantly diff erent. In HPLC, the most polar compounds, like betanin and isobetanin, eluted before less polar decarboxylated and dehydrogenated derivatives (mostly degradation products of betanin). In IP-HSCCC, the dehydrogenated derivatives were eluted faster then their non-dehydrogenated analogues. It was observed for the fi rst time that betanin and neobetanin (14,15-dehydrogenated betanin), which are present in many plants containing betalains, had reversed elution orders during chromatographic separation by these two techniques.