Structural Implications On Color, Xuorescence, and Antiradical Activity in Betalains
Structural Implications On Color, Xuorescence, and Antiradical Activity in Betalains
Structural Implications On Color, Xuorescence, and Antiradical Activity in Betalains
DOI 10.1007/s00425-010-1191-0
O R I G I N A L A R T I CL E
Received: 11 March 2010 / Accepted: 3 May 2010 / Published online: 14 May 2010
© Springer-Verlag 2010
123
450 Planta (2010) 232:449–460
was also investigated from other sources (Butera et al. niWcance of the diVerent structural features is also studied
2002; Pavlov et al. 2002; Cai et al. 2003), and the capacity in terms of structure–antiradical activity relationships.
of betacyanins to inhibit the peroxidation of linoleic acid
and the oxidation of LDL was demonstrated (Kanner et al.
2001). The inhibition of skin and liver tumor formation in Materials and methods
mice has been demonstrated with very low concentrations
of dietary pigments (Kapadia et al. 2003). More recently, Chemicals
protection against the eVects of gamma radiation has also
been reported in these animals (Lu et al. 2009). In addition, All chemicals and reagents were obtained from Sigma (St.
plasma concentrations of betalains after ingestion are suY- Louis, MO, USA). Solvents were from Merck Chemicals
cient to promote their incorporation into the LDL and red Ltd. (Dorset, England). HPLC-grade acetonitrile was pur-
cells, which are then protected from oxidative damage and chased from Labscan Ltd. (Dublin, Ireland). Distilled water
hemolysis in humans (Tesoriere et al. 2003). The potential was puriWed using a Milli-Q system (Millipore, Bedford,
of betalains is in contrast to the lack of systematic analyses MA, USA).
of these molecules, and it is necessary to identify the struc-
tural clues responsible for their biological activities. Extraction and semi-synthesis of betalains
Structurally, betaxanthins are deWned as immonium con-
densation products of betalamic acid with amino acids or The betalains dopaxanthin, betanidin and betanin were
amines. Traditionally, betacyanins have been deWned as extracted from yellow Xowers of Lampranthus productus
condensation products of betalamic acid with cyclo-DOPA, (Gandía-Herrero et al. 2005b), violet Xowers of L. produc-
usually glycosylated (Piattelli 1981). Glycosylation refers tus (Gandía-Herrero et al. 2007), and red roots of Beta vul-
to the multiple possibilities of derivatization of the resulting garis (Wyler and Dreiding 1957), respectively. The rest of
condensation compound, betanidin (Strack et al. 2003). The betalains were obtained as immonium condensation prod-
simplest and best-known glycosylated derivative of betani- ucts of betalamic acid with the following amines: ethylamine,
din is betanin (betanidin-5-O--glucoside). Betacyanin propylamine, 2-phenylethylamine, N-methyl-ethanamine,
deWnition is independent of the actual biosynthetic N-methyl-N-propylamine, pyrrolidine, aniline, N-methyl-
mechanism followed in vivo to reach such a cycle attached aniline, N-ethyl-aniline, indoline, (S)-phenylalanine, and
to the betalamic acid moiety, and there are two plausible (S)-indoline-2-carboxylic acid. Semi-synthesis was carried
pathways proposed in the literature (Piattelli 1981; Gandía- out following a previously described method (Wyler et al.
Herrero et al. 2005b). When structures without the charac- 1965; Gandía-Herrero et al. 2006). In short, betanin puri-
teristic carboxylic group of DOPA were described, they Wed from red beet was used as starting material. Basic
were called descarboxy-betacyanins (Piattelli and Impelliz- hydrolysis (pH 11.4) of 0.2 mM betanin released betalamic
zeri 1970; Kobayashi et al. 2001). In this case, betalamic acid, which was then condensed with the appropriate amine
acid is condensed with descarboxy-cyclo-DOPA, which is after reaching pH 5.0. The corresponding betaxanthin was
cyclo-dopamine. The new pigments possessed the same obtained, revealed by a change of color. The whole process
chromatographic behavior and analogous spectrophotomet- was carried out under nitrogen atmosphere. Once synthesis
rical properties. These were accepted as the only two fami- was achieved, a C-18 solid phase extraction step was per-
lies of betacyanins, leaving the issue of what the structural formed, and an automated system was used for pigment
unit of betacyanins is unresolved. puriWcation.
When betalamic acid is condensed with cyclo-DOPA (or
cyclo-dopamine) to form betacyanins, there is a connection C-18 solid phase extraction
through a proline like substructure between the resonance
system of the betalamic acid moiety and the aromatic sys- One-mL C-18 cartridges (Waters, Milford, MA, USA) were
tem of DOPA. It has been suggested that such a combina- conditioned with 5 mL of methanol followed by 10 mL of
tion is responsible for the particular absorbance properties puriWed water. Salts and buVers from the samples were
of betacyanins. However, there has been no experimental removed by rinsing the column with water. Betaxanthins
approach to the relevance of the diVerent bonds involved in were eluted with ethanol and then concentrated to dryness
the betacyanin structure. under vacuum at room temperature. The residue was redis-
This paper is aimed at exploring the relations between solved in water for further use. The yield obtained for the
the betalain pigment structures and the spectroscopic prop- process was in the range of 92–97%. Sample conductivity
erties they exhibit. DiVerences between betaxanthins and after desaltation was below 1.0 mS/cm for all samples (pH/
betacyanins are investigated, and the minimum structure C-900 conductivity detector, General Electric Healthcare,
responsible for betacyanin properties is identiWed. The sig- Milwaukee, USA).
123
Planta (2010) 232:449–460 451
123
452 Planta (2010) 232:449–460
peroxidase type VI, obtained from Sigma) in the presence structures for the puriWed pigments. For molecules with a
of hydrogen peroxide (45 M), in 12 mM sodium acetate quaternary ammonium substructure (compounds 4, 5, 6, 8,
buVer, pH 5.0. The reactive was then diluted by 2/3 through 9, 10, 12, 14, and 15), the mass obtained corresponds to the
the addition of samples, and the reactions were carried out charged form, as shown in Fig. 1. In all cases a daughter ion
in 53 mM sodium phosphate buVer, pH 7.0. Other condi- corresponding to [M + H]+–44 m/z was detected, account-
tions are speciWed in the text. Measurements of 96-well ing for the loss of a carboxylic group. For compound 15,
plates were performed after 24-h incubations at 20°C in a the main daughter ion (389 m/z) corresponded to compound
Synergy HT plate reader (Bio-Tek Instruments, Winooski, 14. The use of Beta vulgaris as the source of betanin and
USA). All experiments were performed in triplicate and the cleavage of a unit of glucose ([M + H]+–162 m/z) con-
mean values and standard deviations were plotted. Final Wrms the O-glucosidic nature of the bond (Cuyckens and
volume was 300 L (calculated path length = 0.87 cm). Claeys 2004; Gandia-Herrero et al. 2008).
Detector linearity under the assay conditions was conWrmed
(r = 0.999). In each case, errors associated with the results Color diVerences between betaxanthins and betacyanins
provided were calculated on the basis of the residual
standard deviations. Since they are pigments the most important feature of beta-
lains is color. Colorant properties are linked to the electron
resonance system supported by both nitrogen atoms of the
Results and discussion molecule. A full range of color, from yellow to violet, was
obtained with the molecules used in the study (Fig. 2a).
Semi-synthesis and characterization of selected pigments Maximum wavelengths obtained ranged from 467 to
542 nm (Table 2). For the betalains with the simplest struc-
The absence of systematic analyses on the diVerent proper- tural features around the nitrogen atom (structures 1, 2, 3,
ties of betalains, and their relation with the pigments struc- 4, 5, 6, 11, and 13), the maximum wavelength is restricted
tures is justiWed by the lack of a suitable procedure to to the range 467–478 nm, so they are yellow. Those mole-
obtain pure pigments. The application of an improved pro- cules involving aromaticity in resonance with the nitrogen
cedure for the semi-synthesis and puriWcation of betaxant- atoms present a bathochromic shift with maximum wave-
hins (Gandía-Herrero et al. 2006) based on the SchiV base lengths of 507 nm (structure 7), 492 nm (structure 8), and
synthesis (Wyler et al. 1965) allowed us to work with both 489 nm (structure 9), and appear orange in color. Com-
natural and previously unconsidered pigments. A relevant pounds with an analogous aromatic resonance system, but
series of structurally related pigments was obtained in order structurally hindered by an extra intramolecular cycle
to clarify the role of diVerent structural features in betalain (structures 10, 12, 14, and 15) possess wavelengths ranging
properties. from 524 to 542 nm. This extra bathochromic eVect makes
Figure 1 shows the compounds obtained and puriWed for them violet. Table 2 lists the spectroscopic properties deter-
the present study. Pigments were selected in order to mined for all the pigments, including spectrum width at an
explore the role of additional electronic resonance through absorbance value half of that at the corresponding maxi-
aromaticity in the betalain pigments and the eVects of cycli- mum. This parameter shows how yellow pigments spectra
zation, carboxylation, and quaternary ammonium charge. are sharper than those for the orange and violet betalains.
A combined procedure for semi-synthesis and puriWcation A positive linear relation between the maximum wave-
was applied to obtain both betaxanthins and betacyanins. length and the calculated width in the range studied can be
Compounds 13, 14, and 15 were extracted and puriWed found (r = 0.9558). Spectra with the characteristic shapes
from natural sources. All pigments were analyzed by HPLC for the diVerent groups of pigments identiWed above are
using a PDA detector to conWrm purity of the samples shown in Fig. 2b. In all cases a secondary peak in the UV
obtained. Retention times are shown in Table 1. Elution region accompanies the main absorbance peak in the visible
order was related with polarity, with N-ethyl-aniline region.
derived betalain (compound 9) eluting the last. In the series In addition, molar absorption coeYcients were calcu-
of aromatic compounds, betanin (compound 15) was the lated for all the semi-synthetic betalains. A degradation
Wrst to elute due to the eVect of glucosylation. procedure based on the basic hydrolysis of betalains to
Electrospray ionization mass spectrometry (ESI–MS) betalamic acid was applied, using betanin solutions of
was applied to the characterization of the compounds. The known concentration as reference (65,000 M¡1 cm¡1)
mass values determined for the parent ions of all com- (Schwartz and von Elbe 1980; Trezzini and Zrÿd 1991).
pounds are listed in Table 1. In all cases, values were as Values are high for all the molecules, and varied from
expected for the corresponding protonated molecular ions 46,000 to 76,000 M¡1 cm¡1, as shown in Table 2. Mole-
[M + H]+ of the betalains, thus conWrming the proposed cules with maximum absorbance wavelengths between 467
123
Planta (2010) 232:449–460 453
and 478 nm, possess a molar absorption coeYcient sation can be performed with primary and secondary
restricted to the range 46,000–50,000 M¡1 cm¡1. In con- amines and aminoacids. As a result, there are compounds
trast, those pigments of violet color have the highest molar with betaxanthin properties involving cycles (6), aromatic-
absorption coeYcient values, except for compound 14 ity (3, 11, 13), and with a positive charge on the nitrogen
(betanidin). For the compound derived from (S)-indoline-2- group (4, 5, 6). The data obtained in this study support the
carboxylic acid (compound 12) a molar absorption coeYcient use of the mean value of = 48,000 M¡1 cm¡1 at 480 nm
of 76,000 M¡1 cm¡1 was obtained. This is the betalain with for all betaxanthins (Schliemann et al. 2001). Previous esti-
the highest absorbance ever reported. mations of molar absorption coeYcients for other betaxant-
For pigments 7, 8, and 9, the coeYcient values were hins were similar to the present data (Trezzini and Zrÿd
between those of the yellow and violet pigments. Figure 2c 1991).
shows how the maximum wavelengths and the values for On the other hand, molecules with betacyanin properties
the molar absorption coeYcients are related, and it classi- contain an aromatic system in resonance with the betalamic
Wes the betalains graphically. Three diVerent groups are acid moiety, plus a second cycle fused in an indoline manner.
displayed according to their spectroscopic properties: Compound 10 can be considered the simplest pigment with
Betaxanthins, with restricted values for the variables pre- betacyanin properties, and thus the betacyanin deWning
sented, betacyanins with the highest values in absorbance structure. It is likely that the Wve bonded ring causes a
and wavelengths, and compounds 7, 8 and 9, with interme- structural distortion of the planarity in the resonance system
diate properties. and it may be responsible for the characteristics of the
As can be seen, the inclusion into the yellow betaxanthin derived pigments (compounds 10, 12, 14, and 15). This is
group is independent of the nature of the amine condensed more evident when considering the properties of compound
to the betalamic acid moiety, insofar as there is no aroma- 7. In this case, the resonance system is unrestricted, and
ticity in resonance with the nitrogen atoms. Thus, conden- likely to be fully planar. Even in the absence of a second
123
454 Planta (2010) 232:449–460
Table 1 Chromatographic and mass spectroscopic data for betalains used in this study
Amine Trivial Rt PDA-m [M + H]+ Main daughter References
name (min) (nm)a (m/z)b ion (m/z)
cycle, the introduction of a methyl or an ethyl residue at the anthins. A linear relation can be found in the decrease of
nitrogen level may cause a minor distortion and this may be the Stokes shift with increasing excitation wavelengths
behind the diVerent properties of compounds 8, and 9, (r = 0.9807).
respectively, with respect to compound 7. The width for the spectra at half the maximum intensity
is also given (Table 2). Shapes of the spectra obtained in all
Structural considerations on betalains Xuorescence cases respond to the same pattern for each identiWed group,
and are shown in Fig. 3. For simple betaxanthins, spectra
Fluorescence properties of all the betalains were investi- widths range between 22 and 33 nm for excitation, and
gated. Excitation spectra were obtained by following the between 21 and 23 nm for emission. Those betaxanthins
emission at the maximum emission wavelength determined containing an aromatic ring present excitation and emission
in a previous measurement. Emission spectra were obtained widths of 38–48 nm and 25–31 nm, respectively. For
by exciting at the corresponding maximum wavelength. betacyanin compound 13, spectra widths were calculated
Maximum wavelengths are presented in Table 2. As can be as 38 nm for excitation, and 25 nm for emission. For
seen, betaxanthins present maximum excitation wave- compounds 8 and 9, excitation and emission widths were
lengths between 471 and 474 nm, while emission maxima similar (20–24 nm and 21 nm respectively), while for
are found in the range 548–551 nm. For those betacyanins compound 7, widths are considerably higher (41 nm for
where Xuorescence could be detected, compounds 10 and excitation, and 25 nm for emission).
12, maximum excitation wavelengths were 521 and The inXuence of structural factors on the Xuorescence
529 nm, respectively, and emission maxima occurred at intensity of betalains can be detected from the molecules
570 and 575 nm, respectively. Compounds 7, 8, and 9 pres- studied. The presence of a carboxylic group in the pigment
ent intermediate Xuorescence characteristics with maxi- structures leads to an enhancement of the Xuorescence dis-
mum excitation wavelengths between 494 and 513 nm, and played by pigments 11 and 12 with respect to their decar-
maximum emission wavelengths between 553 and 560 nm. boxylated analogous molecules 3 and 10, respectively. This
Figure 3 shows both excitation and emission spectra for may be related to the electron density withdrawing eVect of
characteristic betalain molecules of the diVerent groups. the group, as was also identiWed in the case of carboxylated
Separation between both spectra maxima (Stokes shift) is tyrosine and DOPA derived betaxanthins with respect to
shown in Table 2, and it was found to be higher for betax- tyramine and dopamine derived pigments (Gandía-Herrero
123
Planta (2010) 232:449–460 455
123
456 Planta (2010) 232:449–460
m m Width m Exc m Ems Stokes Relativ. Flu. Exc. width Ems. width
(nm) (M¡1 cm¡1) (nm)a (nm) (nm) shift (nm) Int. (%)b (nm)a (nm)a
123
Planta (2010) 232:449–460 457
Fig. 4 Contour plots of 3D Xuorescence spectra for all betalains under study. No signal could be detected for compounds 14 and 15. All spectra
were obtained in water at 25°C, at a pigment concentration of 3 M. For comparison purposes, relative units are used
123
458 Planta (2010) 232:449–460
123
Planta (2010) 232:449–460 459
shown earlier, the presence of this carboxylic group Cai Y, Sun M, Corke H (2001) IdentiWcation and distribution of simple
enhances Xuorescence intensity of betalains due to its elec- and acylated betacyanins in the Amaranthaceae. J Agric Food
Chem 49:1971–1978
tron density withdrawing eVect. It is also responsible for a Cai Y, Sun M, Corke H (2003) Antioxidant activity of betalains
bathochromic shift in the absorbance properties of carbox- from plants of the Amaranthaceae. J Agric Food Chem
ylated betalains, increasing the maximum wavelength of 51:2288–2294
their visible spectra. This is the case of the betaxanthin 11 Cai Y-Z, Sun M, Xing J, Luo Q, Corke H (2006) Structure–radical
scavenging activity relationships of phenolic compounds from
with respect to 3, and the betacyanin 12 with respect to 10 traditional Chinese medicinal plants. Life Sci 78:2872–2888
(Table 2). Castellanos-Santiago E, Yahia E (2008) IdentiWcation and quantiWca-
By using carboxylated betalains, derived from amine tion of betalains from the fruits of 10 Mexican prickly pear
moieties of unique (S) conWguration and under the HPLC cultivars by high-performance liquid chromatography and elec-
trospray ionization mass spectrometry. J Agric Food Chem
system capable of isomer separation described, the eVect of 56:5758–5764
indoline ring closure on isomers retention times can be ana- Cuyckens F, Claeys M (2004) Mass spectrometry in the structural anal-
lysed. Table 1 shows the retention times for the form (2S/S) ysis of Xavonoids. J Mass Spectrom 39:1–15
of betalains 11–15. (2S/S) is the main natural conWguration Escribano J, Pedreño MA, García-Carmona F, Muñoz R (1998) Char-
acterization of the antiradical activity of betalains from Beta vul-
for the pigments, but they can also be found as the (2S/R) garis L. roots. Phytochem Anal 9:124–127
forms (Gandía-Herrero et al. 2005b; Sasaki et al. 2009). For Gandia-Herrero F, Lorenz A, Larson T, Graham IA, Bowles DJ, Rylott
these isomers the retention times obtained are 19.54 min EL, Bruce NC (2008) DetoxiWcation of the explosive 2,4,6-trini-
(11), 20.04 min (12), 12.65 min (13), 15.03 min (14), and trotoluene in Arabidopsis: discovery of bifunctional O- and
C-glucosyltransferases. Plant J 56:963–974
12.06 min (15). Retention time seems to be independent of Gandía-Herrero F, Escribano J, García-Carmona F (2005a) Betaxant-
the presence of the ring, and it is not aVected by its closure hins as pigments responsible for visible Xuorescence in Xowers.
for the pair of molecules 3 and 10, while it is shortened in Planta 222:586–593
the case of the pair 11 and 12, and enlarged for the pair 13 Gandía-Herrero F, Escribano J, García-Carmona F (2005b) Betaxant-
hins as substrates for tyrosinase. An approach to the role of tyros-
and 14. However, elution order follows a trend based on the inase in the biosynthetic pathway of betalains. Plant Physiol
conWguration of the ring. For those diasteroisomeric beta- 138:421–432
lains with the closed cycle, the elution order for the isomers Gandía-Herrero F, García-Carmona F, Escribano J (2005c) A novel
is (2S/S) earlier than (2S/R), while for the analogous mole- method using high-performance liquid chromatography with Xuo-
rescence detection for the determination of betaxanthins. J Chro-
cules studied with an open cycle the order is (2S/S) later matogr A 1078:83–89
than (2S/R). The eVect of the indoline ring closure on the Gandía-Herrero F, García-Carmona F, Escribano J (2006) Develop-
elution order supports previous observations on the chro- ment of a protocol for the semi-synthesis and puriWcation of
matographic behavior of betalains (Wybraniec 2007). betaxanthins. Phytochem Anal 17:262–269
Gandía-Herrero F, Escribano J, García-Carmona F (2007) Character-
The results of this work show the implications of beta- ization of the activity of tyrosinase on betanidin. J Agric Food
lain pigment structure in their displayed color, Xuores- Chem 55:1546–1551
cence, and chromatographic behavior, and establish the Gandía-Herrero F, Escribano J, García-Carmona F (2009) The role of
structure–activity relationships in relation to the free radical phenolic hydroxy groups in the free radical scavenging activity of
betalains. J Nat Prod 72:1142–1146
scavenging capacity. This not only explains the diVerent Gliszczyjska-Jwigio A, Szymusiak H, Malinowska P (2006) Betanin,
properties of the pigments, but may also allow the identiW- the main pigment of red beet: molecular origin of its exceptionally
cation of potentially interesting molecules and natural high free radical-scavenging activity. Food Addit Contam
sources. 23:1079–1087
Heuer S, Richter S, Metzger JW, Wray V, Nimtzt M, Strack D (1994)
Betacyanins from bracts of Bougainvillea glabra. Phytochemistry
Acknowledgments This work was supported by AGL2007-65907 37:761–767
(MCINN, FEDER, Spain) and by Programa de ayudas a Grupos de Impellizzeri G, Piattelli M, Sciuto S (1973) A new betaxanthin from
Excelencia de la Región de Murcia, de la Fundación Séneca, Agencia Glottiphyllum longum. Phytochemistry 12:2293–2294
de Ciencia y Tecnología de la Región de Murcia (Plan Regional de Kanner J, Harel S, Granit R (2001) Betalains, a new class of dietary
Ciencia y Tecnología 2007/2010). F. G.-H. holds a contract with the cationized antioxidants. J Agric Food Chem 49:5178–5185
“Programa Ramón y Cajal” (MCINN, FEDER, Spain). The authors Kapadia GJ, Azuine MA, Sridhar R, Okuda Y, Tsuruta A, Ichiishi E,
declare that they have no conXict of interest. Mukainake T, Takasaki M, Konoshima T, Nishino H, Tokuda H
(2003) Chemoprevention of DMBA-induced UV-B promoted,
NOR-1-induced TPA promoted skin carcinogenesis, and DEN-
induced phenobarbital promoted liver tumors in mice by extract
References of beetroot. Pharmacol Res 47:141–148
Kobayashi N, Schmidt J, Wray V, Schliemann W (2001) Formation
Butera D, Tesoriere L, Di Gaudio F, Bongiorno A, Allegra M, Pintaudi and occurrence of dopamine-derived betacyanins. Phytochemis-
AM, Kohen R, Livrea MA (2002) Antioxidant activities of sicil- try 56:429–436
ian prickly pear (Opuntia Wcus indica) fruit extracts and reducing Lu X, Wang Y, Zhang Z (2009) Radioprotective activity of betalains
properties of its betalains: betanin and indicaxanthin. J Agric from red beets in mice exposed to gamma irradiation. Eur J Phar-
Food Chem 50:6895–6901 macol 615:223–227
123
460 Planta (2010) 232:449–460
Moreno DA, García-Viguera C, Gil JI, Gil-Izquierdo A (2008) Beta- Schliemann W, Cai Y, Degenkol T, Schmidt J, Corke H (2001) Beta-
lains in the era of global agri-food science, technology and nutri- lains of Celosia argentea. Phytochemistry 58:159–165
tional health. Phytochem Rev 7:261–280 Schwartz SJ, von Elbe JH (1980) Quantitative determination of indi-
Musso H (1979) Pigments of Xy agaric, Amanita muscaria. Tetrahe- vidual betacyanin pigments by high-performance liquid chroma-
dron 35:2843–2853 tography. J Agric Food Chem 28:540–543
Pavlov A, Kovatcheva P, Georgiev V, Koleva I, Ilieva M (2002) StaVord HA (1994) Anthocyanins and betalains: evolution of the mutu-
Biosynthesis and radical scavenging activity of betalains during ally exclusive pathways. Plant Sci 101:91–98
the cultivation of red beet (Beta vulgaris) hairy root cultures. Stintzing FC, Schieber A, Carle R (2002) IdentiWcation of Betalains
Z Naturforsch 57c:640–644 from Yellow Beet (Beta vulgaris L.) and Cactus Pear [Opuntia
Pedreño MA, Escribano J (2001) Correlation between antiradical Wcus-indica (L.) Mill.] by high-performance liquid chromatogra-
activity and stability of betanine from Beta vulgaris L. roots under phy–electrospray ionization mass spectrometry. J Agric Food
diVerent pH, temperature and light conditions. J Sci Food Agric Chem 50:2302–2307
81:627–631 Strack D, Vogt T, Schliemann W (2003) Recent advances in betalain
Piattelli M (1981) The betalains: structure, biosynthesis, and chemical research. Phytochemistry 62:247–269
taxonomy. In: Conn EE (ed) The biochemistry of plants, vol 7. Tesoriere L, Butera D, D’Arpa D, Di Gaudio F, Allegra M, Gentile C,
Academic Press, New York, pp 557–575 Livrea MA (2003) Increased resistance to oxidation of betalain-
Piattelli M, Impellizzeri G (1970) 2-Descarboxybetanidin, a minor enriched human low density lipoproteins. Free Radic Res 37:689–
betacyanin from Carpobrothus acinaciformis. Phytochemistry 696
9:2553–2556 Trezzini GF, Zrÿd J-P (1991) Characterization of some natural and
Piattelli M, Minale L, Nicolaus RA (1965) Betaxanthins from Mirabi- semisynthetic betaxanthins. Phytochemistry 30:1901–1904
lis jalapa L. Phytochemistry 4:817–823 von Ardenne R, Döpp H, Musso H, Steiglich W (1974) Über das Vork-
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C ommen von MuscaXavin bei Hygrocyben (Agaricales) und seine
(1999) Antioxidant activity applying an improved ABTS radical Dihydroazepin-Struktur. Z Naturforsch 29c:637–639
cation decolorization assay. Free Radic Biol Med 26:1231–1237 Wybraniec S (2007) A method for identiWcation of diastereomers of
Sasaki N, Abe Y, Goda Y, Adachi T, Kasahara K, Ozeki Y (2009) 2-decarboxy-betacyanins and 2,17-bidecarboxy-betacyanins in
Detection of DOPA 4,5-dioxygenase (DOD) activity using reversed-phase HPLC. Anal Bioanal Chem 389:1611–1621
recombinant protein prepared from Escherichia coli cells harboring Wyler H, Dreiding AS (1957) Kristallisiertes Betanin. Helv Chim Acta
cDNA encoding DOD from Mirabilis jalapa. Plant Cell Physiol 40:191–192
50:1012–1016 Wyler H, Wilcox ME, Dreiding AS (1965) Umwandlung eines Betac-
Schliemann W, Kobayashi N, Strack D (1999) The decisive step in yans in ein Betaxanthin. Synthese von Indicaxanthin aus Betanin.
betaxanthin biosynthesis is a spontaneous reaction. Plant Physiol Helv Chim Acta 48:361–366
119:1217–1232
123