Carotenoid Profile Research Articles

An optimal saponification and extraction method to determine carotenoids in avocado

Quantification of carotenoids in avocado fruit is a great challenge due to their co-extraction with high-oil concentration and the inherent nature of carotenoids to degrade and undergo cis/trans photoisomerization with prolonged extraction times and high temperatures. The study provides an optimised and validated methodology for quantification of carotenoids in the high-oil avocado matrix, with > 93% recovery of all carotenoids tested being significantly greater than previously published. Saponification with 15% KOH for 60 min was optimal for the avocado matrix. For the first time, this study identified that soap, produced during the saponification reaction, resulted in a significant reduction of carotenoid content from the avocado matrix, due to the production of micelles. A significantly higher carotenoid content (3.58 versus 2.0 mg/100 g DW) was able to be extracted after saponification with acidified phosphate buffer, instead of water as reported previously. Carotenoid profiles of five avocado cultivars were identified and quantified.

Nanoencapsulation of Red Bell Pepper Carotenoids: Comparison of Encapsulating Agents in an Emulsion Based System

Red bell pepper extract rich in carotenoids was (RBPE) encapsulated with four different encapsulating agents: calcium caseinate (ECC), bovine gelatin (EBG), whey proteins isolate (EWPI), and concentrate (EWPC), aiming to investigate the most effective material to coat and enable the water dispersibility of pigments. Formulations were obtained by the oil in water (O/W) emulsification technique, followed by freeze-drying. Samples were analyzed by encapsulation efficiency, high-performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), atomic force microscopy (AFM), thermogravimetric analysis (TGA), dispersion stability, and CIELab. Nanoformulations showed a carotenoid encapsulation efficiency of 54.0% (ECC), 57.6% (EWPI), 56.6 % (EWPC), 64.0 % (EBG). Recovered carotenoid profiles from nanoformulations showed similarity to the RBPE, indicating the efficiency of the encapsulation process. Average particle sizes of approximately 109 nm (ECC), 71 nm (EWPI), 64 nm (EWPC), and 173 nm (EBG) were obtained. AFM revealed that all formulations exhibited spherical forms and a heterogeneous distribution profile. Regarding TGA, formulations presented similar thermal behaviors to and lower decomposition speeds than RBPE, suggesting improved thermal stability. Powder formulations were easily dispersed in water (8 mg/mL) and presented intense color and stability to sedimentation for 48 h. Results indicated that all formulations and the chosen technique efficiently increased carotenoid dispersibility in water, indicating their potential to be applied as natural food pigments.

Genetic Diversity and Association Analysis for Carotenoid Content among Sprouts of Cowpea (Vigna unguiculata L. Walp).

The development and promotion of biofortified foods plants are a sustainable strategy for supplying essential micronutrients for human health and nutrition. We set out to identify quantitative trait loci (QTL) associated with carotenoid content in cowpea sprouts. The contents of carotenoids, including lutein, zeaxanthin, and β-carotene in sprouts of 125 accessions were quantified via high-performance liquid chromatography. Significant variation existed in the profiles of the different carotenoids. Lutein was the most abundant (58 ± 12.8 mg/100 g), followed by zeaxanthin (14.7 ± 3.1 mg/100 g) and β-carotene (13.2 ± 2.9 mg/100 g). A strong positive correlation was observed among the carotenoid compounds (r ≥ 0.87), indicating they can be improved concurrently. The accessions were distributed into three groups, following their carotenoid profiles, with accession C044 having the highest sprout carotenoid content in a single cluster. A total of 3120 genome-wide SNPs were tested for association analysis, which revealed that carotenoid biosynthesis in cowpea sprouts is a polygenic trait controlled by genes with additive and dominance effects. Seven loci were significantly associated with the variation in carotenoid content. The evidence of variation in carotenoid content and genomic regions controlling the trait creates an avenue for breeding cowpea varieties with enhanced sprouts carotenoid content.

Influence of cultivar and season on carotenoids and phenolic compounds from red lettuce influence of cultivar and season on lettuce

This paper presents complete HPLC profiles and MS spectrometric data of bioactive compounds from four cultivars of red lettuce produced in winter and summer and their antioxidant capacity. The experiment was carried out in a greenhouse, where red curly lettuce was cultivated: Mila, Maira, Carmin and Scarlet. The cultivar and season have not influenced the qualitative profile of carotenoids (CAR) and phenolic compounds (PC) of red lettuce. Instead, the season influenced the concentration of these components in all cultivars. The levels of phenolic compounds were significantly higher in winter, while the levels of carotenoids were higher in summer. Ten anthocyanins were identified (cyanidins and delphinidins). The main carotenoid found was the all-trans-β-carotene (45–48%), followed by lutein (13–20%) and zeaxanthin (11–15%). Major phenolic compounds include 5-caffeoylquinic acid, rutin and amentoflavone. Red lettuce cultivars have their main bioactive compounds described and compared within the variety and within the growing season. Different season and different lettuce cultivars may differ in the content of their bioactive compounds and in their antioxidant capacity.

Comparative Characteristics of Carotenoid Profiles and Antiradical Properties of Extracts of Brown Kelp from the Sea of Japan

Comparative Characteristics of Carotenoid Profiles and Antiradical Properties of Extracts of Brown Kelp from the Sea of Japan

Morphological and Biochemical Variation in Carrot Genetic Resources Grown under Open Field Conditions: The Selection of Functional Genotypes for a Breeding Program

Carrot (Daucus carota), one of the most economically important root vegetables, shows a wide range of morphological and biochemical diversity. However, there is a lack of simultaneous systematic study regarding the biochemical composition and morphological characteristics in carrot genetic resources, which is crucial for crop improvement. For this reason, the morphological characteristics, carotenoids, and free sugar content of 180 carrot genetic resources grown in open field conditions from March to June 2020 were accessed to select the lines for a potential breeding program. Altogether, 15 qualitative and 4 quantitative agronomical characteristics were evaluated and grouped into four categories based on root color (orange, yellow, white, and purple). Three carotenoids (lutein, α-carotene, and β-carotene) and three free sugars (fructose, glucose, and sucrose) were also analyzed. The results revealed wide genetic variation in both qualitative and quantitative traits. Most of the genetic resources were orange (n = 142), followed by white (n = 16), yellow (n = 14), and purple (n = 8). Carotenoid profile and content were highly dependent on root color and showed wide genetic variability, while sugar content and profile were independent of the root color. Alpha- and β- carotene were the major carotenoids in orange carrots representing 43.3 and 41.0% of total carotenoids. In contrast, lutein was most dominant in other colored carrots (79.7–98.6% of total carotenoids). In most of the genetic resources, sucrose was the most dominant free sugar, followed by glucose and fructose. The results of this study showed that some genetic resources elevated carotenoid and sugar content. The morphological and biochemical diversity observed in this study might be useful for improving the agronomic traits and biochemical content of carrot lines for breeding programs.

Map-Based Cloning and Characterization of Br-dyp1, a Gene Conferring Dark Yellow Petal Color Trait in Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

Flower color is an important trait in Brassica species. However, genes responsible for the dark yellow flower trait in Chinese cabbage have not been reported. In this study, we identified a dark-yellow-flowered Chinese cabbage line SD369. Genetic analysis indicated that the dark yellow flower trait in SD369 was controlled by a single recessive locus, Br-dyp1 (dark yellow petal color 1 in Brassica rapa). Using bulked segregant RNA sequencing and kompetitive allele-specific PCR assays, Br-dyp1 was fine-mapped to an interval of 53.6 kb on chromosome A09. Functional annotation analysis, expression analysis, and sequence variation analysis revealed that Bra037130 (BraA09.ZEP), which encodes a zeaxanthin epoxidase, was the most likely candidate gene for Br-dyp1. Carotenoid profile analysis suggested that Bra037130 (BraA09.ZEP) might participate in the epoxidation from zeaxanthin to violaxanthin. The 679 bp insertion in dark yellow petal caused premature stop codon, thus caused the loss-of-function of the enzyme zeaxanthin epoxidase (ZEP), which disturbed the carotenoid metabolism, and caused the increased accumulation of total carotenoid, and finally converted the flower color from yellow to dark yellow. Comparative transcriptome analysis also showed that the “carotenoid biosynthesis” pathway was significantly enriched, and genes involved in carotenoid degradation and abscisic acid biosynthesis and metabolism were significantly downregulated. Furthermore, we developed and validated the functional marker Br-dyp1-InDel for Br-dyp1. Overall, these results provide insight into the molecular basis of carotenoid-based flower coloration in B. rapa and reveal valuable information for marker-assisted selection breeding in Chinese cabbage.

Transgene-free genome editing supports CCD4 role as a negative regulator of β-carotene in banana.

This study aims to understand the regulatory mechanism of the β-carotene homeostasis by establishing transgene-free genome editing in banana. Carotenoid cleavage dioxygenases (CCDs) belong to a miniature gene family having an imperative role in the intricated carotenoid metabolism in plants. Here, the expression pattern of multiple CCDs was correlated with the levels of carotenoid accumulation in two contrasting cultivars, viz., Nendran (high β-carotene) and Rasthali (low β-carotene). The higher expression of the RAS-CCD4 inversely correlated with β-carotene accumulation in fruit-pulp of the Rasthali. The docking analysis followed enzyme assay of purified RAS-CCD4 suggested β-carotene and 10-apo-β-carotenal as its preferred substrates. Bacterial complementation assay affirmed RAS-CCD4 role in β-carotene degradation and then overexpression of the RAS-CCD4 in the Arabidopsis thaliana further validated results in-vivo by the significant reduction in β-carotene. Subsequently, CRISPR/Cas9 mediated editing of CCD4 was demonstrated in the protoplasts and embryogenic cell lines of Rasthali. The carotenoid profiling in stable mutant lines revealed higher fold β-carotene accumulation in non-green tissue (roots) than in green tissue (leaf) compared with the unedited control plants. The differential expression of carotenoid pathway genes was correlated with the metabolites in the edited lines. The study suggests that carotenoid catabolism is regulated by the CCD4 in tissue and cultivar specific manners, and also demonstrated the use of the genome editing tool in developing transgene-free biofortified banana.

Extraction of astaxanthin from Haematococcus pluvialis with hydrophobic deep eutectic solvents based on oleic acid

Three novel hydrophobic deep eutectic solvents (DESs) based on oleic acid and terpenes (thymol, dl-menthol, and geraniol) were prepared, characterized, and used to extract astaxanthin from the microalga Haematococcus pluvialis without any pre-treatment of the cells. The three DES were composed of Generally Recognized As Safe (GRAS) and edible ingredients. All the tested DESs gave astaxanthin recovery values of about 60 and 30% in 6 h if applied on freeze-dried biomass or directly on algae culture, respectively. The carotenoid profile was qualitatively identical to what was obtained by using traditional organic solvents, regardless of the DES used; the monoesters of astaxanthin with C18-fatty acids were the main compounds found in all the carotenoid extracts. The thymol:oleic acid DES (TAO) could preserve astaxanthin content after prolonged oxidative stress (40% of the astaxanthin initially extracted was still present after 13.5 h of light exposure), thanks to the superior antioxidant properties of thymol. The capacity of improving astaxanthin stability combined with the intrinsic safety and edibility of the DES components makes the formulation astaxanthin-TAO appealing for the food ingredients/additives industry.

First complete quali-quantitative carotenoids characterization of Aiphanes aculeata, Quararibea cordata and Garcinia intermedia fruits

Several tropical fruits are important sources of bioactive molecules. The carotenoid composition of three fruits, Corozo, Sastra and Yellow sapote, from the area of Panama, was investigated in order to provide for the first time the complete quali-quantitative carotenoid profile. The carotenoid content in Yellow sapote was related to the fruit size, being higher in smaller fruits. Zeaxanthin was one of the main carotenoids in all fruits, and very high amounts of β-cryptoxanthin and β-carotene were determined in Sastra and Corozo (respectively, 70.8 μg/g and 32.2 μg/g in Sastra and 181.9 μg/g and 74.7 μg/g in Corozo); moreover, Corozo showed also a very high amount of lycopene (45.2 μg/g). Considering the here determined high carotenoids content and the fact that the investigated fruits are little commercialized and found in areas where vitamin A deficiency is still widespread, their production and use as source of bioactives should be much improved.

Overexpression of OsMYBR22/OsRVE1 transcription factor simultaneously enhances chloroplast-dependent metabolites in rice grains

The OsMYBR22 (same to OsRVE1), an R1type-MYB transcription factor belonging to the rice CCA1-like family, was upregulated under blue light condition, which enhanced the chlorophyll and carotenoid accumulation. The overexpression of OsMYBR22 in rice (Oryza sativa, L) led to everlasting green seeds and leaves of a darker green. Transgene expression patterns showed more concordance with chlorophyll than carotenoid profiles. The transcript levels of most genes related to chlorophyll biosynthesis and degradation examined were similarly repressed in the late maturing stages of seeds. It proposed that rice seeds have the feedback regulatory mechanism for chlorophyll biosynthesis and also implied that evergreen seed traits might be caused due to the inhibition of degradation rather than the promotion of biosynthesis for chlorophylls. Metabolomics revealed that OsMYBR22 overexpression largely and simultaneously enhanced the contents of nutritional and functional metabolites such as chlorophylls, carotenoids, amino acids including lysine and threonine, and amino acid derivatives including γ-aminobutyric acid, which are mostly biosynthesized in chloroplasts. Transmission electron microscopy anatomically demonstrated greener phenotypes with an increase in the number and thickness of chloroplasts in leaves and the structurally retentive chloroplasts in tubular and cross cells of the seed inner pericarp region. In conclusion, the molecular actions of OsMYBR22/OsRVE1 provided a new strategy for the biofortified rice variety, an “Evergreen Rice,” with high accumulation of chloroplast-localized metabolites in rice grains.

Characterization of the Antioxidant Activity, Carotenoid Profile by HPLC-MS of Exotic Colombian Fruits (Goldenberry and Purple Passion Fruit) and Optimization of Antioxidant Activity of this Fruit Blend

Characterization of the Antioxidant Activity, Carotenoid Profile by HPLC-MS of Exotic Colombian Fruits (Goldenberry and Purple Passion Fruit) and Optimization of Antioxidant Activity of this Fruit Blend

Effects of herbivory on carotenoid biosynthesis and breakdown.

The carotenoid content of plants may be impacted by stress with major consequences for photosynthesis and photoprotection. Most carotenoid stress research, however, has concentrated on abiotic stresses, and we know little about how biological stresses, such as herbivory, alter profiles of plant carotenoids and their degradation products. For example, carotenoid derivatives such as β-cyclocitral and β-ionone have been recently shown to act as signals in plant growth and protection against oxidative stress and herbivory. To understand how carotenoid composition is influenced by herbivory, changes in biosynthesis and degradation should be investigated. This chapter describes methods to simulate herbivory in a simple reproducible fashion and to assess carotenoid biosynthesis and degradation. Carotenoid biosynthesis depends on precursors provided by the methylerythritol 4-phosphate (MEP) pathway, which converts pyruvate and glyceraldehyde-3-phosphate to the five-carbon units used for construction of larger isoprenoids. We present protocols to quantify the activity of the first enzyme of the MEP pathway, deoxy-xylulose 5-phosphate synthase (DXS), usually assumed to be rate-controlling, and to estimate the concentration of the first intermediate of the pathway, deoxy-xylulose 5-phosphate (DXP). We also discuss procedures to measure the formation of volatile carotenoid breakdown products after herbivory. To monitor the activity of carotenoid-specific biosynthetic enzymes, such as phytoene synthase, protocols are available elsewhere in this volume (Wurtzel, 2022b).

Chemical composition of caferana (Bunchosia armeniaca) fruit: a rich source of lycopene

The present study aimed to evaluate the nutritional quality of the caferana fruit, focusing on the content and profile of carotenoids due to the characteristic coloring of these compounds. The fruits were collected, botanically identified, and evaluated for proximate composition, carotenoid and sugar content and profile, and flavonoid profile. The results of the centesimal composition of the caferana pulp were: 65 g/100g of moisture; 0.72 g/100g of ash; 0.92 g/100g of protein; 0.78 g/100g of ethereal extract; 2.83 g/100g of dietary fiber; 29.73 g/100g of carbohydrates and the caloric value of 129.62 kcal/100g of pulp. As for the content and profile of sugars in the pulp, 19.41 g/100g of fructose and 4.01 g/100g of glucose were obtained. The fruit had a high content of carotenoids (40 mg/100g on a wet basis), of which 89% was lycopene (36 mg/100g), 2.5 mg/100g of β-carotene and 0.3 mg/100g of lutein. Rutin was identified as a flavonoid present in the fruit pulp. The caferana fruit proved to be an excellent source of lycopene, with higher levels than tomato-based products.

The breeder's tool-box for enhancing the content of esterified carotenoids in wheat: From extraction and profiling of carotenoids to marker-assisted selection of candidate genes.

Carotenoid esterification is a new target for cereal biofortification since esterification increases both accumulation and stability of carotenoids. A xanthophyll acyl transferase is responsible for carotenoid esterification in the endosperm of wheat and related cereals. In this chapter we describe the procedures for transferring the carotenoid esterification attribute into wheat using the wild barley Hordeum chilense as donor of the esterification trait, the outline of the breeding program and the protocols for marker assisted selection and the analysis of carotenoids in grain. Biofortified cereals with increased lutein ester content will help to reduce the risk of developing age-related macular degeneration in human populations with limited access to other dietary sources.

Understanding carotenoid biosynthetic pathway control points using metabolomic analysis and natural genetic variation.

Advances in analytical methodologies, including those coupled with mass spectrometry (gas chromatography and liquid chromatography; GC-MS and LC-MS) have facilitated profiling of carotenoids in complex plant extracts. The combination of metabolomic data together with diverse germplasm resources provides a means to discover the underlying genetic factors responsible for modulating the carotenoid biosynthetic pathway. Here we summarize metabolomics methodologies for large-scale metabolite analysis and provide guidelines for genetic analysis to leverage natural variation for identification of genes encoding biosynthetic pathway enzymes based on linkage mapping and/or genome-wide association studies (GWAS). We also demonstrate the workflow for identifying genes in carotenoid biosynthesis based on natural variation using case studies in several species.

Ethanol Associated with Ultrasound Efficiently Extracted Carotenoids from Peels of Peach Palm Fruits (Bactris Gasipaes Kunth) Without Altering Carotenoid Profile

Ethanol Associated with Ultrasound Efficiently Extracted Carotenoids from Peels of Peach Palm Fruits (Bactris Gasipaes Kunth) Without Altering Carotenoid Profile

HPLC analysis of carotenoids from bacteria.

Bacteria synthesize different carotenoid structures. A powerful method for the determination of carotenoid profiles of bacteria, including quantification, is high performance liquid chromatography (HPLC). This chapter explains the basics of this method and its application, and provides instruction on how to proceed with HPLC. A basic HPLC system for bacterial carotenoids is presented which allows tentative identification and quantification. The individual analysis steps including extraction, sample preparation and HPLC separation are described in detail. HPLC examples of structurally diverse carotenoids from different bacterial phyla are displayed demonstrating the applicability of this method. In addition, information on spectroscopic properties useful for identification is included. Finally, a procedure for carotenoid quantification from the HPLC chromatogram is described.

Optimization of Culture Conditions for Total Carotenoid Amount Using Response Surface Methodology in Green Microalgae / Ankistrodesmus convolutus

Commercial carotenoids of green microalgae have become significant especially for their applications in the cosmetic, pharmaceutical, food and feed industries. Effects of physical and chemical parameters on carotenoid contents in isolated microalgal species have been investigated. The variables of shaking rate, nitrogen concentration and light intensity affect biomass production and the synthesis of carotenoids in the green microalgae were investigated using the statistical design by Box-Behnken (BBD) employing Response Surface Methodology (RSM). Furthermore, the optimized cultivation conditions using BBD for Chlorella vulgaris, Ankistrodesmus convolutus, Dunaliella salina, Tetraselmis striata were determined using the spectrophotometric method to enhance carotenoid concentration. A. convolutus within the green algae was detected with the highest carotenoid concentration. The optimum conditions results indicated that the growth of A. convolutus (0.55 mg/L) and production of total carotenoids (25.1138 mg/g biomass) were found at the stirrer rate of 100 rpm under the light intensity of 100 μE/m2 s, and in the nutrient component of 8.82 mM NaNO3 . These conditions were validated experimentally for total carotenoid yield (24.13 mg/g biomass). After that the production was performed in a flat-plate photobioreactor with a volume of 6L based on the optimized conditions and the carotenoid profile was defined by HPLC-DAD using standards such as violaxanthin, astaxanthin and β-carotene. This study proposes that the RSM approach can be used to define optimal conditions for large-scale production of carotenoids by A. convolutus. 

Hypercarotenemia in Anorexia Nervosa Patients May Influence Weight Balance: Results of a Clinical Cross-Sectional Cohort Study.

Introduction: Anorexia nervosa (AN) can co-occur with hypercarotenemia, a clinical condition characterized by elevated β-carotene in plasma and skin tissue. Carotenoids have known anti-obesogenic effects in adipocyte biology. Thus, carotenoids may potentially play a retarding role in weight gain during the recovery of AN patients. This study evaluated the plasma carotenoid profile and subcutaneous adipose tissue (SAT) in a cohort of AN patients and normal weight (NW) controls.Methods: Plasma concentrations of α-carotene, β-carotene, β-cryptoxanthin, and lycopene were determined by HPLC analysis. SAT thicknesses were measured by a highly accurate and reliable ultrasound technique. Information on dietary intakes were collected by repeated 24-h recalls.Results: Sixty-two females (AN: n = 18, NW: n = 44) were included. The concentrations of β-cryptoxanthin (p = 0.045) and lycopene (p = 0.004) were significantly lower in AN patients. β-carotene levels were higher in AN patients (n.s.) and α-carotene did not differ significantly. SAT thickness was significantly lower in AN patients compared to controls (p < 0.001). β-carotene was significantly negative (rs = −0.471) and lycopene significantly positive (rs = 0.366) correlated with SAT. The correlation of β-carotene and SAT was even higher in the AN group alone (rs = −0.742). Also, β- cryptoxanthin and the sum of provitamin A carotenoids were correlated to SAT (rs = −0.647 and rs = −0.746, respectively) in AN patients. Fruits and vegetable intake did not differ significantly between AN and NW but adjusted for SAT, AN patients consumed relatively higher amounts (p = 0.006).Conclusion: Higher plasma β-carotene concentrations were associated with reduced SAT levels, most probably due to a reduced ability of the remaining adipose tissue to store carotenoids. Thus, the antiobesity effects of carotenoids might impact the treatment success of undernutrition and AN. A systemic carotenoid overload may contribute to changes in adipogenesis and metabolic capacities for energy storage. Therefore, high plasma β-carotene may be a marker of delay in weight recovery in AN patients. Interventional studies should consider including carotenoid-status in AN treatment.