Black Carrot Anthocyanins Research Articles

Improvement of storage stability and bioaccessibility of microencapsulated black carrot (Daucus Carota ssp. sativus) anthocyanins using maltodextrin and sericin protein combinations as wall material

This study explores the potential of microencapsulation to enhance the stability and bioaccessibility of anthocyanins from black carrots (Daucus carota ssp. sativus). Anthocyanins, known for their health benefits, are often limited in their application due to poor stability and bioaccessibility. We investigated microencapsulation using maltodextrin alone (BAM) and in combination with sericin proteins (BAMS1, BAMS2, BAMS3), a novel approach. Spray drying was employed to encapsulate anthocyanins, with maltodextrin (20%) and varying concentrations of sericin protein (1%, 2%, 3%) as wall materials. Comprehensive analysis encompassed parameters such as production yield, moisture content, bulk and tapped density, encapsulation efficiency, particle size distribution, surface morphology, FTIR, amino acid profiling, storage stability under diverse temperature conditions (4 °C, 25 °C, 37 °C), and simulated gastrointestinal conditions to assess bioaccessibility. Results indicated a range of production yield (51.05%–59.13%) and encapsulation efficiency (87.68%–94.01%), with moisture content between 5.24% and 6.47%. Sericin incorporation notably influenced moisture content and particle density. Particle size varied from 74.14 μm to 160 μm, with discernible differences attributed to sericin concentration. SEM revealed smooth-continuous surface morphologies, indicating the influence of sericin on capsule structure. Preservation of essential amino acid profiles underscored the efficacy of the encapsulation process. Enhanced stability and degradation kinetic parameters of anthocyanins, particularly evident in BAMS3 microcapsules, underscored the protective role of sericin. In vitro digestion studies confirmed controlled release dynamics, highlighting the potential for improved anthocyanin bioaccessibility facilitated by the sericin-maltodextrin composite encapsulation approach.

Stability of individual anthocyanins from black carrots stored in light and darkness – Impact of acylation

Black carrot anthocyanins have gained increasing attention as natural coloring agent, owing to their higher stability than anthocyanins from berries. The stability has been attributed to their higher degree of acylation. This study investigated the impact of acylation on the stability of individual anthocyanins during storage in light and darkness. We hypothesized that the acylated anthocyanins would be more stable than the non-acylated ones. The major five anthocyanins were fractioned by semi-preparative HPLC and stored at pH 4.5 in light and darkness to investigate how acylation affected the stability. The stability was evaluated by absorption spectroscopy and mass spectrometry (MS). Two of the anthocyanins were non-acylated; 3-xylosyl(glucosyl)galactoside and cyanidin 3-xylosylgalactoside, and three were acylated; cyanidin 3-xylosyl(sinapolyglucosyl)galacto-side, cyanidin 3-xylosyl(feruloylglu-cosyl)galactoside, and cyanidin 3-xylosyl(coumaroyl-glucosyl)galactoside. Both methods (spectroscopy and MS) showed a clear effect of acylation when stored in light, but surprisingly the two non-acylated anthocyanins, showed higher stability than the three acylated ones.

Effect of storage on physicochemical attributes of ice cream enriched with microencapsulated anthocyanins from black carrot.

The present study was conducted to investigate the effect of storage on quality attributes of microencapsulated black carrot anthocyanins-enriched ice cream. Purposely, black carrot anthocyanins were obtained using ethanolic extraction. Later on, extracts were acidified and microencapsulated with gum arabic and maltodextrin (1:1). Results showed that anthocyanin contents for T3 (9% microencapsulated anthocyanins powder-enriched ice cream) had highest anthocyanin contents in the range of 143.21 ± 1.14 mg/100 g. However, during the storage, it was revealed that there was a slight decline in constituents concentration reasoned to oxygen exposure and interaction with other food ingredients. Similarly, for total phenolic content, the highest amount was found in T3 as 545.38 ± 4.34 mg GAE/100 g. The quality attributes of prepared ice cream treatments were also found acceptable till the end of the study (60 days). Conclusively, the addition of microencapsulated anthocyanins powder in ice cream proved to stabilize black carrot anthocyanins and contributed positively to the sensory characteristics of ice cream.

Entrapment of Black Carrot Anthocyanins by Ionic Gelation: Preparation, Characterization, and Application as a Natural Colorant in Yoghurt

Black carrot (BC) with its potential health benefits due to the greater amount of anthocyanins and the potent antioxidant activity could be utilized as a natural colorant. The objective of this study was the entrapment of BC anthocyanins by external ionic gelation technique within the biopolymer matrix including pectin, alginate, and the mixture of both. Beads were characterized in terms of entrapment efficiency (EE), morphology, total anthocyanin content, and antioxidant capacity measured by the 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid assay. Furthermore, the color of the beads as well as yoghurt samples fortified with BC-containing beads were evaluated during storage at 4 °C for 4 weeks. While the EE for anthocyanins ranged between 47.3 and 96.6%, the antioxidant capacity changed from 50.4 to 97.7%. The maximum anthocyanin retention was found as 91.7% for 1% BC containing 1% pectin (P) + 1% alginate (A)-based beads after 4 weeks of storage. In addition, anthocyanin protection reached up to 62% and antioxidant capacity up to 55.6% in the fortified yoghurt samples containing A-based beads during storage. It is concluded that external ionic gelation could be a feasible method for BC anthocyanins due to its protective effect against acidic environment.

Stability enhancement of beetroot betalains through copigmentation.

Like other natural pigments, betalains have a stability problem. Copigmentation can be explored to address this issue. In this study, black carrot anthocyanins were used for the first time as copigment with betalains so that copigmented betalains with enhanced stability could be developed to withstand deteriorative processing and storage conditions. Increase in hyperchromic and bathochromic shift with subsequent increase in black carrot anthocyanin extract (0.250 g L-1 ) addition from 0.2 to 1.0mL L-1 was observed in native betalain pigments from 0.28 to 1.90 and 538 nm to 564 nm, respectively. For maximum recorded bathochromic shift, 0.8mL L-1 addition of copigment was optimized. Copigmented betalain pigment showed better stability in comparison with native pigment, when exposed to light, temperature more than 60 °C and ≥1.0g L-1 NaCl. At constant incubation time (3h), copigmented betalains degraded up to 20.79-41.43% whereas the non-copigmented counterpart degraded up to 83.49-86.86% at 60, 75 and 90 °C, respectively. Lower rate constant (k) and enhanced activation energy (Ea ) showed higher thermostability of copigmented betalains. With constant light exposure, the half-life value of betalains was 145.2h, which increased approximately twofold (274.08 h) after copigmentation. The t1/2 of betalain pigment at 10%, 15% and 18% salt addition was 81.12, 75.36 and 83.52 h, which increased to 186.96, 226.56 and 152.88 h after copigmentation. These findings support that black carrot anthocyanin is a potential and compatible copigment for water-soluble betalain pigment that enhances stability of betalains under extreme processing conditions. © 2022 Society of Chemical Industry.

Black carrot anthocyanins exhibit neuroprotective effects against MPP+ induced cell death and cytotoxicity via inhibition of oxidative stress mediated apoptosis.

The online version contains supplementary material available at 10.1007/s10616-021-00500-4.

The effect of whey protein concentration and preheating temperature on the color and stability of purple corn, grape and black carrot anthocyanins in the presence of ascorbic acid

Our objective was to explore the effects of whey protein (WP) concentration and preheating temperature on anthocyanin color expression and stability over time in the presence of ascorbic acid. Anthocyanins from purple corn, grape or black carrot were mixed with native WP or preheated WP (40–80°C) in various concentrations (0–10 mg/mL) in pH 3 buffer containing 0.05% ascorbic acid and stored in the dark at 25 °C for 5 days. WP addition increased anthocyanin absorbance and protected anthocyanin from ascorbic acid-mediated degradation. Increasing WP concentration resulted in lower lightness and higher chroma, hue angle and color stability. The color loss of anthocyanin solutions decreased by 40%-50% when 10 mg/mL WP was added. Native WP showed more color enhancement and protection than thermally-induced WP. Increasing the WP preheating temperature resulted in less absorbance increase and more absorbance loss. Anthocyanin’ half-life was improved by addition of WP in a dose dependent manner. Native WP addition (10 mg/mL) extended anthocyanin half-life by about 2 times for purple corn and grape, and 1.31 times for black carrot anthocyanin solutions. Preheating temperature did not significantly affect anthocyanin protection by WP. WP addition might enhance anthocyanin stability in beverages containing ascorbic acid, expanding anthocyanin application in foods.

A Roadmap to Modulated Anthocyanin Compositions in Carrots.

Anthocyanins extracted from black carrots have received increased interest as natural colorants in recent years. The reason is mainly their high content of acylated anthocyanins that stabilizes the color and thereby increases the shelf-life of products colored with black carrot anthocyanins. Still, the main type of anthocyanins synthesized in all black carrot cultivars is cyanidin limiting their use as colorants due to the narrow color variation. Additionally, in order to be competitive against synthetic colors, a higher percentage of acylated anthocyanins and an increased anthocyanin content in black carrots are needed. However, along with the increased interest in black carrots there has also been an interest in identifying the structural and regulatory genes associated with anthocyanin biosynthesis in black carrots. Thus, huge progress in the identification of genes involved in anthocyanin biosynthesis has recently been achieved. Given this information it is now possible to attempt to modulate anthocyanin compositions in black carrots through genetic modifications. In this review we look into genetic modification opportunities for generating taproots of black carrots with extended color palettes, with a higher percentage of acylated anthocyanins or a higher total content of anthocyanins.

Colouring properties and stability of black carrot anthocyanins in yoghurt.

Among the natural pigments anthocyanins have potential to be applied as natural colourants besides exhibiting wide range of bioactivity. Colouring potential and storage stability of black carrot concentrate (BCC) containing anthocyanins in yoghurt was determined in present investigation. The instrumental colour (CIELAB) values were altered by the addition of BCC in yoghurt in which the L* and b* values decreased, while a* value increased with increasing levels of BCC. Maximum sensory scores were observed for yoghurt with 1.5% BCC, as it was similar to strawberry in colour and appearance. Enhancement in the total anthocyanin, total phenolics and DPPH antioxidant activity was observed with increasing levels of BCC in yoghurt. L* value remained same during storage in both yoghurts, but a* value increased slightly. Similar trend was also noticed in BCC yoghurt for anthocyanins and antioxidant activity. The total phenolic content got enhanced in control, but decreased significantly in BCC yoghurt. Sensory evaluation revealed that scores decreased during storage but the product was acceptable up to 15days. Our study further confirmed that higher stability and better colouring properties of black carrot concentrate in fermented milk system was due to higher degree of acylation.

Characterization of black carrot (Daucus carota L.) polyphenols; role in health promotion and disease prevention: An overview

This era is witnessing a drastic shift towards increasing the consumption of functional foods for promoting healthy lifestyle. Functional foods are recognized because of their high nutraceutical properties, which helps in prevention of various degenerative disorders. Black carrot is considered as functional food because of the presence of significant amount of bioactive compounds. The present review primarily aims at spotlighting the presence of potential phytochemicals in black carrots (Daucus Carota L.) and their significant role in oxidative stress induced major metabolic disorder. Effect of processing on polyphenols and anthocyanins of black carrot are also discussed. Black carrot anthocyanins are considered the major bioactive compound, which is associated with its health promotion and disease prevention properties. Functional properties of these bioactive compounds call for further studies to exploit its role in prevention of diseases.

Carrot Anthocyanins Genetics and Genomics: Status and Perspectives to Improve Its Application for the Food Colorant Industry.

Purple or black carrots (Daucus carota ssp. sativus var. atrorubens Alef) are characterized by their dark purple- to black-colored roots, owing their appearance to high anthocyanin concentrations. In recent years, there has been increasing interest in the use of black carrot anthocyanins as natural food dyes. Black carrot roots contain large quantities of mono-acylated anthocyanins, which impart a measure of heat-, light- and pH-stability, enhancing the color-stability of food products over their shelf-life. The genetic pathway controlling anthocyanin biosynthesis appears well conserved among land plants; however, different variants of anthocyanin-related genes between cultivars results in tissue-specific accumulations of purple pigments. Thus, broad genetic variations of anthocyanin profile, and tissue-specific distributions in carrot tissues and organs, can be observed, and the ratio of acylated to non-acylated anthocyanins varies significantly in the purple carrot germplasm. Additionally, anthocyanins synthesis can also be influenced by a wide range of external factors, such as abiotic stressors and/or chemical elicitors, directly affecting the anthocyanin yield and stability potential in food and beverage applications. In this study, we critically review and discuss the current knowledge on anthocyanin diversity, genetics and the molecular mechanisms controlling anthocyanin accumulation in carrots. We also provide a view of the current knowledge gaps and advancement needs as regards developing and applying innovative molecular tools to improve the yield, product performance and stability of carrot anthocyanin for use as a natural food colorant.

Designing a pH-responsive drug delivery system for the release of black-carrot anthocyanins loaded in halloysite nanotubes for cancer treatment

Anthocyanins extracted from black-carrot were encapsulated into halloysite nanotubes (HNT) as a potential carrier for the first time. Anthocyanins were loaded into HNT under vacuum conditions. TEM images indicated the successful entrance of anthocyanins into the lumen of HNT. Anthocyanins loading efficiency was determined to be 4% using thermogravimetric analysis (TGA). XRD patterns showed that all reflections shifted to higher values for HNT loaded with anthocyanins compared to pure HNT which indicates reduced d-value of the crystalline forms. In vitro release of anthocyanins was studied under gastrointestinal conditions using the pH-differential method. The drug release from HNT was modeled and based on R2 values, the Korsmeyer-Peppas model was introduced as the best model predicting the release behavior. Cytotoxic effects of the samples against MCF-7 human breast cancer and HT-29 colon cancer cell lines were evaluated using SRB assay. The cell viability was reduced by 2 folds against anthocyanins-loaded HNT compared to pure anthocyanins in both cell lines. Anthocyanins-loaded HNT is suggested as a promising pH-responsive drug delivery system for the treatment of cancer.

A novel pH-sensing indicator based on bacterial cellulose nanofibers and black carrot anthocyanins for monitoring fish freshness

A novel intelligent pH-sensing indicator based on bacterial nanocellulose (BC) and black carrot anthocyanins (CA) was developed and characterized to monitor the freshness/spoilage of rainbow trout and common carp fillet during the storage at 4 °C. The indicator displayed wide color differences from red to gray over the 2–11 pH range, which was clearly discerned by the naked eye. The fabricated pH-sensing indicator showed distinguishable color changes during fresh (deep carmine color), best to eat (charm pink color), and spoiled (jelly bean blue and khaki colors) stages of both fish fillets. Moreover, a strong and positive correlation was obtained between the total color differences values of the indicator and bacterial count (R = 0.952 and 0.991) and total volatile basic nitrogen (R = 0.815 and 0.92) in rainbow trout and common carp samples. The results of this work demonstrated a significant correspondence of fish shelf life and color changes of a nanocellulose-based pH-sensing indicator.

Intelligent Chitosan/PVA nanocomposite films containing black carrot anthocyanin and bentonite nanoclays with improved mechanical, thermal and antibacterial properties

This study investigated the potential of chitosan/PVA films containing black carrot anthocyanins as a natural pH indicator and bentonite as a nano-filler, for intelligent packaging applications. Two series of chitosan/PVA (CP) films, with different concentrations of bentonite (CPB3%, CPB5%) and with a constant concentration of anthocyanin (CPA, CPAB3%, CPAB5%) were prepared by solution casting method. The colorimetric parameter b* showed an increase from 1.0 ± 0.2 for CP films to 2.7 ± 0.2 and 8.8 ± 1.1 for CPB3% and CPB5% films, respectively which indicates that addition of bentonite increases the intensity of yellow color in the nanocomposite films. The presence of anthocyanins caused to shifting of a* values from negative to positive values which indicated the color change of films from greenness to redness. Addition of bentonite reduced the tensile strength from 100.6 for CP film to 86.5 and 67.2 for CPB3% and CPB5% films which was related to the solution intercalation method used for preparation of nanocomposite films. In contrast, addition of anthocyanins increased the tensile strength for CPA, CPAB3% and CPAB5% due to the hydrogen bonding interactions between its OH groups and PVA and chitosan chains. Water vapor permeability (WVP) of the films was decreased from 1.140 (mg.mm/m2.kPa.h) for CP film to 0.5620 and 0.3515 for CPB3% and CPB5% films, respectively. In contrast, anthocyanins caused to increase WVP of the samples compared to the anthocyanin free films. Thermo-gravimetric analysis showed that the films containing bentonite and anthocyanins have a better thermal stability. Finally, CPAB5% showed the highest bacterial growth inhibition against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, respectively 75.20%, 69.95% and 44.38%.

Formation and characterization of spray dried coated and uncoated liposomes with encapsulated black carrot extract

The degradation of black carrot anthocyanins (BCE) under stress conditions as under the effect of a spray drying process was evaluated in different encapsulation systems (extract, chitosan-coated and uncoated liposomes and chitosan-extract-mixture). The encapsulation efficiencies of liposomes prepared by high-pressure homogenization containing 2% lecithin with 0.1%, 0.2%, and 0.4% extract were found as 86.6% ± 16.1%, 82.2% ± 9.7%, and 46.9% ± 6.3%, respectively. The extract-loaded liposomes (0.2% extract) were chosen for liposomes considering further dilutions. The chitosan-coated liposomes were generated with 0.1% chitosan according to their particle size (82.7 ± 1.5 nm) and zeta potential of 51.4 ± 0.6 mV. All samples were found to be physically stable after the spray drying process. In addition, SEM images of the liposomal powder particles showed a uniformly wrinkled surface that would indicate stability of liposomal systems against spray drying. However, the liposomal samples showed lower stability compared to the BCE and BCE-chitosan mixtures according to the biochemical analysis of the reconstituted powders.

Black Carrot (Daucus carota ssp.) and Black Soybean (Glycine max (L.) Merr.) Anthocyanin Extract: A Remedy to Enhance Stability and Functionality of Fruit Juices by Copigmentation

Application of extracted black carrot anthocyanins (BCA) and black soybean anthocyanins (BSA) as colour enhancers was demonstrated by copigmenting with heat sensitive anthocyanins from plum, pomegranate, strawberry and jamun. Improved pigment stability was confirmed by thermal stability, chromacity studies and hyperchromic and bathochromic shifts. BCA (2%, 500 mg/L) caused largest hyperchromic shift in plum whereas maximum bathochromic shift (18–20 nm) was observed in strawberry. BSA (2%, 500 mg/L) caused largest hyperchromic shift in pomegranate whereas maximum bathochromic shift (22 nm) was observed in plum. Copigmented plum, jamun, strawberry and pomegranate juices copigmented with BCA achieved high half-life (t1/2) and lower rate constant (k) values. Copigmenting anthocyanins with other juice is an effective strategy to impart high thermal stability, improve colour retention and antioxidant activity.

Synthesis of silica-PAMAM dendrimer nanoparticles as promising carriers in Neuro blastoma cells

Mesoporous silica carriers are emerging as therapeutic drug delivery systems. The objective of this study was to develop a formulation for synthesizing silica-PAMAM dendrimer hybrid nanoparticles with sol-gel technique. Subsequently, black carrot anthocyanins were encapsulated and investigated for their capability in terms of inhibiting the proliferative effects of neuroblastoma (Neuro 2A). In this context, particle size distributions were ascertained followed by thermal analysis (DSC), scanning electron microscopy and encapsulation efficiency. Subsequently, in vitro release kinetics was determined along with cytotoxicity of empty and anthocyanin doped hybrid nanoparticles. The lowest particle size was 134.8 nm with a zeta potential of +19.78 mV which enhanced electrostatic interaction with the cell membrane in the cytotoxicity analyses. As the anthocyanin content was totally released at the end of 6 days, the cytotoxicity was observed for 134 h, reaching an inhibition of 87.9%. On the other hand, Neuro 2A cells incubated with empty nanoparticles exhibited a high proliferation indicating that hybrid nanoparticles were not toxic to the cells and the inhibitory effect was associated with the anthocyanins.

Green Extraction Processes Focusing on Maximization of Black Carrot Anthocyanins along with Cytotoxic Activities

Interest in anthocyanins has intensified due to their health promoting ability by reducing the risk of atherosclerosis, cancer, diabetes, and neurodegenerative disorders. In this study, anthocyanin extraction from black carrot was optimized by using solvent, ultrasound-, and microwave-assisted techniques, where the independent variables were temperature, liquid/solid ratio, and extraction time. The extractions were carried out with ethanol, whereas water was applied in each technique under elicited optimum conditions. Total anthocyanin content of the extracts was determined by a pH differential method, and the extracts possessing the highest anthocyanin contents were further analyzed with UPLC. Additionally, cytotoxic activity of the extracts was tested against breast adenocarcinoma, bone osteosarcoma, alveolar adenocarcinoma, and neuroblastoma. Microwave-assisted extraction yielded the highest amount of anthocyanins (319.57 μg/mL) when a liquid/solid ratio of 5:1 was applied at 60 °C for only 9 min. The cytotoxic activity was remarkable in most of the cell lines with IC50 values below 5.5 μg/mL. Consequently, the results obtained results suggest that microwave-assisted extraction of black carrot has a potential to be scaled up for full utilization on an industrial scale.

Effect of genuine non-anthocyanin phenolics and chlorogenic acid on color and stability of black carrot (Daucus carota ssp. sativus var. atrorubens Alef.) anthocyanins

This work aimed at studying the color intensity and stability of black carrot anthocyanins as influenced by intermolecular co-pigmentation. For this purpose, purified anthocyanin solutions were supplemented with purified genuine black carrot phenolics, chlorogenic acid, and an aqueous phenolic-rich green coffee bean extract at various anthocyanin:co-pigment ratios (1:0–1:162; pH 3.6). The hyperchromic co-pigmentation effect depended on the concentration of added co-pigments, resulting in an absorbance increase of up to 22% at the absorption maximum. Anthocyanin stability during heating (90°C, 5h) was barely improved unless the concentrations of co-pigments exceeded those of their natural source. When adding co-pigments at ratios above 1:9.4, anthocyanin heat stability was significantly improved. As acylated anthocyanins were most stable, breeders might aim at increasing their content in the future, while breeding for high levels of colorless polyphenols may be unreachable. Nevertheless, we provided proof-of-concept for the successful color enhancement by the addition of a phenolic-rich green coffee bean extract, being useful for food-grade applications.

Optimization of Extraction of Bioactive Compounds from Black Carrot Using Response Surface Methodology (RSM)

In this study, optimum conditions for the extraction of black carrot anthocyanins were determined by response surface methodology. Central composite design of extraction factors (pH 2.5–6.5, temperature 4–72 °C, solvent/solid ratio 5:1–25:1 v/w, ethanol/water ratio 0:100–100:0 v/v) was generated as two replicates. Total phenolic content, total monomeric anthocyanin content, polymeric color, total antioxidant activity, and anthocyanin composition determined by high-performance liquid chromatography were used as responses. Except for color analysis, higher temperature, solid/solvent ratio, and ethanol concentration were observed to increase the extraction yield. However, polymeric color results were found to have minimum values at lower pH and solid/solvent ratio, lower or moderate temperature, and higher ethanol concentration. Optimum extraction conditions were found as follows: 50 °C, pH 3.5, solvent solid ratio 10:1 (v/w), and ethanol/water ratio 75:25 (v/v) when all responses were considered. The validation of the optimum conditions for black carrot extraction was performed at specified values.