Supercritical extraction of betalains from the peel of different pitaya species.

BACKGROUND: Pitaya is a fruit with high consumer acceptance and health benefits. Pitaya peel is a waste product with potential in the food industry, as an antioxidant enrichment and natural colouring. Therefore, there is an interest in recovering its constituents and searching for pitaya species with greater potential. This work aimed to obtain bioactive extracts from the dried peel of pitaya fruits of the species Selenicereus monacanthus (Lem.), S. costaricensis W. and S. undatus H. using supercritical fluids at different pressures (100, 250 and 400 bar) and ethanol-water 15% v/v or ethanol 100% as co-solvents. The extraction yield, antioxidant activity, colour and total betalain content were evaluated. RESULTS: The extract obtained from S. monacanthus showed the highest extraction yield (49.6 g kg-1), followed by S. costaricensis (27.5 g kg-1) and S. undatus (17.7 g kg-1) at 400 bar and 35 °C using ethanol 15%, v/v. The antioxidant capacity was strongly influenced by pressure, favouring the obtaining of betalain-rich extracts at higher pressures, especially in the species S. costaricensis (0.6 g kg-1) and S. monacanthus (0.3 g kg-1). To improve the extraction of S. undatus (the most cultivated species), the procedure of subsequential extractions was applied. This procedure considerably increased the extraction yield, antioxidant activity and total content of betalains. The use of ethanol 100% provided more bioactive fractions and achieved a good separation of betalains. CONCLUSION: The supercritical extraction method can overcome the challenge of efficiently extracting compounds from pitaya peel, due to the presence of bioactive compounds of great polarity. © 2024 Society of Chemical Industry.

Extraction and Stabilization of Betalains from Beetroot (Beta vulgaris) Wastes Using Deep Eutectic Solvents.

Deep eutectic solvents (DES) using magnesium chloride hexahydrate [MgCl2·6H2O] and urea [U] proportions (1:1) and (2:1), were prepared for their use as extracting and stabilizer agents for red and violet betalains from beetroot (Beta vulgaris) waste. The synthetized DES [MgCl2·6H2O] [U] showed similar properties to eutectic mixtures, such as, liquid phase, low melting points and conductivity, thermal stability, and variable viscosity. In turn, betalain DES extracts (2:1) exhibited compatibility in the extraction and recovery of betalains from beetroot wastes, showing a betalain content comparable to that of betalain extracts. Betalain stability was determined by degradation tests; the exposure conditions were visible light (12 h), molecular oxygen from atmospheric air and environmental temperature (20-27 °C) for 40 days. The kinetic curves of the betalain degradation of water samples depicted a first-order model, indicating the alteration of a violet colouration of betalains from beetroot waste for 5-7 days. However, betalains from DES extracts were kept under visible light for 150 days, and for 340 days in storage (amber vessels), achieving a stability of 75% in comparison with initial beet extracts.

Optimisation of the ultrasound-assisted extraction of betalains and polyphenols from Amaranthus hypochondriacus var. Nutrisol.

The present study optimised the ultrasound-assisted extraction (UAE) of bioactive compounds from Amaranthus hypochondriacus var. Nutrisol. Influence of temperature (25.86-54.14 °C) and ultrasonic power densities (UPD) (76.01-273.99 mW/mL) on total betalains (BT), betacyanins (BC), betaxanthins (BX), total polyphenols (TP), antioxidant activity (AA), colour parameters (L*, a*, and b*), amaranthine (A), and isoamaranthine (IA) were evaluated using response surface methodology. Moreover, betalain extraction kinetics and mass transfer coefficients (KLa) were determined for each experimental condition. BT, BC, BX, TP, AA, b*, KLa, and A were significantly affected (p < 0.05) by temperature extraction and UPD, whereas L*, a*, and IA were only affected (p < 0.05) by temperature. All response models were significantly validated with regression coefficients (R2) ranging from 87.46 to 99.29%. BT, A, IA, and KLa in UAE were 1.38, 1.65, 1.50, and 29.93 times higher than determined using conventional extraction, respectively. Optimal UAE conditions were obtained at 41.80 °C and 188.84 mW/mL using the desired function methodology. Under these conditions, the experimental values for BC, BX, BT, TP, AA, L*, a*, b*, KLa, A, and IA were closely related to the predicted values, indicating the suitability of the developed quadratic models. This study proposes a simple and efficient UAE method to obtain betalains and polyphenols with high antioxidant activity, which can be used in several applications within the food industry.

Bioactive Betalain Extracts from Cactus Pear Fruit Pulp, Beetroot Tubers, and Amaranth Leaves.

Natural food items and the additional benefits they provide have received considerable attention in recent years. Betalains are nutritious pigments which have valuable biological properties, e.g., antioxidant and free radical scavenging activity. In this study, aqueous betalain extracts were obtained from different coloured cactus pears (purple, red/pink, and orange), amaranth, and beetroot, with and without the addition of ascorbic acid, microwave-heated, and freeze-dried and subsequently analysed by thin layer chromatography (TLC). Beetroot samples without the addition of ascorbic acid (AA) had lower phenols, flavonoids, and ascorbic acid content than beetroot samples extracted with the addition of AA. Amaranth had significantly higher contents of antioxidants than all the other plants. Results for phenolic compounds showed that there were no significant differences between cactus pear cultivars, however, significant differences were seen between the two beetroot samples (microwave-heated with and without AA) as well as amaranth. For flavonoid compounds, amaranth had significantly higher values than all other samples. The lowest flavonoid content was found in beetroot without AA (0.49 mgCE/g). For ascorbic acid, significant differences were noticed between amaranth (71.71 mg/100 g) and samples from cactus pear and beetroot. TLC results showed that purple and red cactus pear samples had the most vivid colours, a reflection of the high betacyanin and betaxanthin contents in the cultivars. Moreover, extracts from cactus pear, beetroot, and amaranth were classified according to a decision tree which was designed by the Code of Federal Regulations/Food Additives Regulation of the EU. The classification of betalain pigment extracts as colouring foods was achieved through enrichment factor calculations and the colourant decision tree. The results showed that the betalain pigment extraction method used is inexpensive, time-saving, energy-saving, non-toxic, and chemical solvent free and yields high concentrations of betalains.

The Influence of Different Pretreatment Methods on Color and Pigment Change in Beetroot Products.

Vegetable processing pomace contains valuable substances such as natural colors that can be reused as functional ingredients. Due to a large amount of water, they are an unstable material. The aim of our research was to assess how the pretreatment method (thermal or nonthermal) affects the properties of powders obtained from beet juice and pomace after the freeze-drying process. The raw material was steamed or sonicated for 10 or 15 min, and then squeezed into juice and pomace. Both squeezed products were freeze-dried. The content of dry substance; L*, a*, and b* color parameters; and the content of betalain pigments were analyzed. Pretreatments increased the proportion of red and yellow in the juices. Steam and ultrasound caused a significant reduction in parameter b* in the dried pomace. A significant increase in betanin in lyophilizates was observed after pretreatment with ultrasound and steam for 15 min. As a result of all experiments, dried juices and pomaces can also be used as a colorant source. However, there is higher potential with pomaces due to their additional internal substances as well as better storage properties. After a few hours, juice was sticky and not ready to use.

Development of an aqueous ultrasound-assisted extraction process of bioactive compounds from beet leaves: a proposal for reducing losses and increasing biomass utilization.

BACKGROUND: Red beet plants are cultivated worldwide for the consumption of their roots, generating large amounts of unexploited by-products. In particular, beet leaves (BLs) represent about 50% of the whole plant and are usually discarded as waste. This constitutes not only an economic issue, since multiple resources invested in the production will be wasted, but also an environmental problem because of the pollution associated with their disposal. However, BLs comprise an important source of functional compounds (polyphenols and betalains) that could be recovered from the raw material, representing a sustainable solution for the underutilization of this by-product. This study proposes the recovery of polyphenols and betalains using an aqueous ultrasound-assisted extraction (UAE) process at different powers (35, 50, and 100 W) that was characterized and optimized. RESULTS: UAE significantly enhanced the recovery of bioactive compounds and shortened the time required for extraction in comparison with traditional macerations (35 < 50 < 100 W). During UAE, the temperature of the systems increased as a function of the power applied, favouring the recovery of these phytochemicals. Additionally, a Box-Behnken design and response surface methodology were employed to optimize UAE conditions (90 W ultrasound power, 1:20 solid:liquid ratio, 16 min extraction time), under which the yields were 14.9 mg g-1 (polyphenols), 949.1 µg g-1 (betaxanthins), and 562.2 µg g-1 (betacyanins), consistent with the values predicted by the models. CONCLUSION: This study enabled the development of a green-solvent UAE process that constitutes an effective post-harvest by-products strategy to minimize losses and increase biomass utilization through the recovery of bioactive compounds from BLs, promoting sustainability in the agri-food chain. © 2020 Society of Chemical Industry.

Phytochemical Molecules from the Decarboxylation of Gomphrenins in Violet Gomphrena globosa L.-Floral Infusions from Functional Food.

Herein, the generation of decarboxylated derivatives of gomphrenin pigments exhibiting potential health-promoting properties and the kinetics of their extraction during tea brewing from the purple flowers of Gomphrena globosa L. in aqueous and aqueous citric acid solutions were investigated. Time-dependent concentration monitoring of natural gomphrenins and their tentative identification was carried out by LC-DAD-ESI-MS/MS. The high content of acylated gomphrenins and their principal decarboxylation products, 2-, 15-, 17-decarboxy-gomphrenins, along with minor levels of their bidecarboxylated derivatives, were reported in the infusions. The identification was supported by the determination of molecular formulas of the extracted pigments by liquid chromatography coupled with high-resolution mass spectrometry (LCMS-IT-TOF). The influence of plant matrix on gomphrenins' stability and generation of their derivatives, including the extraction kinetics, was determined by studying the concentration profiles in the primary and diluted infusions. Isolated and purified acylated gomphrenins from the same plant material were used for the preliminary determination of their decarboxylated derivatives. The acylated gomphrenins were found to be more stable than nonacylated ones. Citric acid addition had a degradative influence on natural gomphrenins mainly during the longer tea brewing process (above 15 min); however, the presence of plant matrix significantly increased the stability for betacyanins' identification.

Red Beetroot Betalains: Perspectives on Extraction, Processing, and Potential Health Benefits.

In recent years, red beetroot has received a growing interest due to its abundant source of bioactive compounds, particularly betalains. Red beetroot betalains have great potential as a functional food ingredient employed in the food and medical industry due to their diverse health-promoting effects. Betalains from red beetroot are natural pigments, which mainly include either yellow-orange betaxanthins or red-violet betacyanins. However, betalains are quite sensitive toward heat, pH, light, and oxygen, which leads to the poor stability during processing and storage. Therefore, it is necessary to comprehend the impacts of the processing approaches on betalains. In this review, the effective extraction and processing methods of betalains from red beetroot were emphatically reviewed. Furthermore, a variety of recently reported bioactivities of beetroot betalains were also summarized. The present work can provide a comprehensive review on both conventional and innovative extraction techniques, processing methods, and the stability of betalains.

Application of Natural Pigments in Ordinary Cooked Ham.

The possibility of obtaining a carmine or pink color on ordinary cooked ham by applying natural dyes from three plant species, namely red radish (Raphanus sativus L.), hibiscus (Roselle sabdariffa L.) and red beetroot (Beta vulgaris L.), was investigated. The extracts were evaluated for the stability at physical-chemical parameters and subjected to cytotoxicity assays in the gastric cell line AGS Encapsulation of the extracts in soybean lecithin liposomes and maltodextrin microcapsules was performed. Lyophilized extracts before and after encapsulation in maltodextrin were applied in the formulation of ordinary cooked ham and used in a pilot scale of production. The color of cooked ham samples from different assays was evaluated visually and by colorimetry. The results suggest that the coloration of ordinary cooked ham obtained with extracts of red beetroot is very promising for future applications in this type of meat product.

Influence of gelation on the retention of purple cactus pear extract in microencapsulated double emulsions.

A purple cactus pear (Opuntia ficus-indica) extract (CP) was encapsulated in double emulsions (DE) gelled with gelatin (DE-CP-G) and with gelatin and transglutaminase (DE-CP-GT), as well as in a DE with a liquid external aqueous phase (DE-CP), in order to study the retention of betanin as colorant agent. Both gelled DEs showed a predominantly elastic behavior, in contrast with DE-CP. The degradation rate constant of betanin was significantly higher in DE-CP-GT (90.2 x 10-3 days-1) than in DE-CP-G (11.0 x 10-3 days-1) and DE-CP (14.6 x 10-3 days-1) during cold-storage (4 °C). A shift towards yellow color was found in all the systems during cold-storage (4 °C) and after thermal treatment (70°C/30 min), especially in DE-CP-GT, denoting a higher degradation of betanin. Betalamic acid, cyclo-Dopa 5-O-ß-glucoside, 17-decarboxy-betanin and neobetanin were identified by UHPLC-MS/MS as degradation products of betanin.

Red Beetroot and Betalains as Cancer Chemopreventative Agents.

Carcinogenesis is the process whereby a normal cell is transformed into a neoplastic cell. This action involves several steps starting with initiation and followed by promotion and progression. Driving these stages are oxidative stress and inflammation, which in turn encompasses a myriad of aberrant gene expressions, both within the transforming cell population and the cells within the surrounding lesion. Chemoprevention of cancer with bioreactive foods or their extracted/purified components occurs via normalizing these inappropriate gene activities. Various foods/agents have been shown to affect different gene expressions. In this review, we discuss whereby the chemoprevention activities of the red beetroot itself may disrupt carcinogenesis and the activities of the water-soluble betalains extracted from the plant.

Chemistry Inspired by the Colors of Fruits, Flowers and Wine.

An overview is provided of the status of research at the frontiers of investigation of the chemistry and photochemistry of two classes of natural plant pigments, the anthocyanins and the betalains, as well as of the pyranoanthocyanin pigments formed from anthocyanins during the maturation of red wine. Together, anthocyanins and betalains are responsible for almost all of the red, purple and blue colors of fruits and flowers and anthocyanins and pyranoanthocyanins are major contributors to the color of red wines. All three types of pigments are cationic below about pH 3, highly colored, non-toxic, reasonably soluble in water or alcohol and fairly stable to light. They exhibit good antioxidant or antiradical activity and, as part of our diet, confer a number of important health benefits. Systematic studies of model compounds containing the basic chromophoric groups of these three types of pigments are providing a deeper understanding of the often complex chemistry and photochemistry of these pigments and their relationship to the roles in vivo of these pigments in plants. These natural pigments are currently being exploited as starting materials for the preparation of novel semi-synthetic dyes, pigments and fluorescence probes.

Study on Betalains in Celosia cristata Linn. Callus Culture and Identification of New Malonylated Amaranthins.

Betacyanins and betaxanthins were characterized and determined in an intensely pigmented red-colored callus culture of Celosia cristata L. (Amaranthaceae). A new malonyl derivative, 6'- O-malonyl-amaranthin (celoscristatin) was isolated and identified by spectroscopic and mass spectrometric techniques. Its stereoisomer, 4'- O-malonyl-amaranthin (celoscristatin acyl-migrated), as well as its 15 R diastereomer were also detected in the callus as a result of the malonyl group migration in celoscristatin/isoceloscristatin, respectively. Amaranthin occurs in the callus as the major betacyanin, followed by celoscristatin, betanin, phyllocactin, and other minor betacyanins. The effect of different carbon sources on the growth rates of the Celosia callus as well as on betalains profiles in the callus cultures was studied. High dopamine content in the callus culture was determined and compared with the content in C. cristata inflorescences. The dopamine-based betalain (miraxanthin V) was detected as the main betaxanthin in the callus, however, at a concentration level much lower than that of the identified betacyanins. The studied callus culture of C. cristata can accumulate betalains in amounts which approach the quantities produced by most known high-yielding plant species.

Chemistry Inspired by the Colors of Fruits, Flowers and Wine

ABSTRACT An overview is provided of the status of research at the frontiers of investigation of the chemistry and photochemistry of two classes of natural plant pigments, the anthocyanins and the betalains, as well as of the pyranoanthocyanin pigments formed from anthocyanins during the maturation of red wine. Together, anthocyanins and betalains are responsible for almost all of the red, purple and blue colors of fruits and flowers and anthocyanins and pyranoanthocyanins are major contributors to the color of red wines. All three types of pigments are cationic below about pH 3, highly colored, non-toxic, reasonably soluble in water or alcohol and fairly stable to light. They exhibit good antioxidant or antiradical activity and, as part of our diet, confer a number of important health benefits. Systematic studies of model compounds containing the basic chromophoric groups of these three types of pigments are providing a deeper understanding of the often complex chemistry and photochemistry of these pigments and their relationship to the roles in vivo of these pigments in plants. These natural pigments are currently being exploited as starting materials for the preparation of novel semi-synthetic dyes, pigments and fluorescence probes.

Visible light absorption and photo-sensitizing properties of spinach leaves and beetroot extracted natural dyes.

Herein, chlorophyll and betalain dyes are extracted from fresh spinach leaves and beetroots. Fourier transform infrared spectra are used to identify the characteristic peaks of the extracted dyes. UV-vis light absorption characteristics of the dyes and their mixed counterpart are investigated by varying their pH and temperature. These dyes are used as photo sensitizer for fabrication of zinc oxide photo-anode based dye sensitized solar cells (DSSCs). The photo-voltaic characteristics of the developed DSSCs are measured under simulated solar light (power of incident light 100 mW cm(-2) from Air Mass 1.5G). The solar to electric conversion efficiencies for the chlorophyll, betalain and mixed dye based solar cells are estimated as 0.148%, 0.197% and 0.294% respectively. The highest conversion efficiency for mixed dye based solar cell is attributed due to the absorption of wider range of solar spectrum.

Versatile solvent systems for the separation of betalains from processed Beta vulgaris L. juice using counter-current chromatography.

Two mixtures of decarboxylated and dehydrogenated betacyanins from processed red beet roots (Beta vulgaris L.) juice were fractionated by high performance counter-current chromatography (HPCCC) producing a range of isolated components. Mixture 1 contained mainly betacyanins, 14,15-dehydro-betanin (neobetanin) and their decarboxylated derivatives while mixture 2 consisted of decarboxy- and dehydro-betacyanins. The products of mixture 1 arose during thermal degradation of betanin/isobetanin in mild conditions while the dehydro-betacyanins of mixture 2 appeared after longer heating of the juice from B. vulgaris L. Two solvent systems were found to be effective for the HPCCC. A highly polar, high salt concentration system of 1-PrOH-ACN-(NH4)2SO4 (satd. soln)-water (v/v/v/v, 1:0.5:1.2:1) (tail-to-head mode) enabled the purification of 2-decarboxy-betanin/-isobetanin, 2,17-bidecarboxy-betanin/-isobetanin and neobetanin (all from mixture 1) plus 17-decarboxy-neobetanin, 2,15,17-tridecarboxy-2,3-dehydro-neobetanin, 2-decarboxy-neobetanin and 2,15,17-tridecarboxy-neobetanin (from mixture 2). The other solvent system included heptafluorobutyric acid (HFBA) as ion-pair reagent and consisted of tert-butyl methyl ether (TBME)-1-BuOH-ACN-water (acidified with 0.7% HFBA) (2:2:1:5, v/v/v/v) (head-to-tail mode). This system enabled the HPCCC purification of 2,17-bidecarboxy-betanin/-isobetanin and neobetanin (from mixture 1) plus 2,15,17-tridecarboxy-2,3-dehydro-neobetanin, 2,17-bidecarboxy-2,3-dehydro-neobetanin and 2,15,17-tridecarboxy-neobetanin (mixture 2). The results of this research are crucial in finding effective isolation methods of betacyanins and their derivatives which are meaningful compounds due their colorant properties and potential health benefits regarding antioxidant and cancer prevention. The pigments were detected by LC-DAD and LC-MS/MS techniques.

Extraction, stability, and separation of betalains from Opuntia joconostle cv. using response surface methodology.

Betalains were extracted and analyzed from Opuntia joconostle (the prickly pear known as xoconostle in Mexico). For the extraction, two solvent systems were used, methanol/water and ethanol/water. A three-variable Box-Behnken statistical design was used for extraction: solvent concentration (0-80%, v/v), temperature (5-30 °C), and treatment time (10-30 min). The extraction and stability of betalains from xoconostle were studied using response surface methodology (RSM). Techniques such as UV-vis, column chromatography, and HPLC were employed for the separation and analysis of the main pigments present in the extracts. Maximum pigment concentration (92 mg/100 g of fruit) was obtained at a temperature of 15 °C and a time of 10 min for methanol/water (20:80), whereas maximum stability of the pigment was observed at pH 5 and a temperature of 25 °C. HPLC chromatograms showed the main betalains of the xoconostle characterized were betalain, betanidin, and isobetalain.

Purification and antiradical properties of the structural unit of betalains.

Betalamic acid [4-(2-oxoethylidene)-1,2,3,4-tetrahydropyridine-2,6-dicarboxylic acid] is a naturally occurring compound that is normally found condensed with amino acids, amines, cyclo-DOPA, and cyclo-DOPA derivatives to form the betalains. Betalains are the pigments responsible for the yellow to violet color of the fruits and flowers of plants belonging to the order Caryophyllales. Betalamic acid is the structural feature common to all of these pigments and contains the electron resonance system responsible for the spectroscopic properties. Betalamic acid was purified by chromatography and identified by UV-vis spectrophotometry and ESI mass spectrometry. The antioxidant and free radical scavenging capacities of betalamic acid were assessed using the FRAP and ABTS(·+) radical assays. A pK(a) of 6.8 was found for the deprotonation equilibrium involved in the nucleophilic activity of betalamic acid; this pK(a) explains the observed pH effect on the free radical scavenging capacity of these pigments.

Beetroot betalain inhibits peroxynitrite-mediated tyrosine nitration and DNA strand cleavage.

Two major betalains, red-purple betacyanins and yellow betaxanthins, were isolated from red beetroots (Beta vulgaris L.), and their peroxynitrite (ONOO(-)) scavenging capacity was investigated. Apparent colours of the betalains were bleached by the addition of ONOO(-), and the absorbance decreases were suppressed in the presence of glutathione, a ONOO(-) scavenger. After bleaching, a new absorption maximum was observed at 350 nm in the spectrum of the resulting reaction mixture. New peaks were detected from HPLC analysis of the reaction products of betanin, a representative constituent of red beetroot betacyanins, treated with ONOO(-) monitoring at 350 nm, and the intensity of the major peak was positively correlated with ONOO(-) concentration. Betanin inhibited the ONOO(-) (0.5 mM)-dependent nitration of tyrosine (0.1 mM). Additionally, the IC(50) value of betanin (19.2 µM) was lower than that of ascorbate (79.6 µM). The presence of betanin (0.05-1.0 mM) also inhibited ONOO(-) (0.5 mM)-dependent DNA strand cleavage in a concentration-dependent manner. These results suggest that betalains can protect cells from nitrosative stress in addition to protecting them from oxidative stresses.