Effect of Ultrasound-Assisted Extraction of Carotenoids from Papaya (Carica papaya L. cv. Sweet Mary) Using Vegetable Oils

By-products from fruits and are of great interest for their potential use in the food industry due to their high content of bioactive compounds. Herein, we examined the ultrasound-assisted extraction (UAE) of carotenoid and carotenoid esters from papaya pulp and peel using soybean oil and sunflower oil as alternative green solvents. Response surface methodology (RSM) was established to optimize the UAE process. Three independent variables, ultrasonic amplitude (20–60%), time (10–60 min), and co-solvent percentage (ethanol) (5–20%, v/v), were applied. The highest total carotenoid content in the UAE extracts was obtained from papaya pulp extracts (58.7 ± 1.6 and 56.0 ± 1.5 μg carotenoids/g oil) using soybean oil and sunflower oil, respectively (60% amplitude/ 10 min/ 20% ethanol). On the other hand, the highest carotenoid content (52.0 ± 0.9 μg carotenoids/g oil) was obtained from papaya peel using soybean oil applying the UAE process (20% amplitude/ 77 min/ 20% ethanol); a minor content of 39.3 ± 0.5 μg carotenoids/g oil was obtained from papaya peel using sunflower oil at 60% amplitude/ 60 min/ 5% ethanol. Lycopene was the most abundant carotenoid among all individual carotenoids observed in papaya oil extracts, obtaining the highest yields of this carotenoid when papaya pulp and peel were extracted using soybean oil (94% and 81%, respectively) and sunflower oil (95% and 82%, respectively). Great extraction of xanthophyll esters was detected using 20% of ethanol in the vegetable oil extraction solvent (v/v). High correlations (>0.85) was obtained between total carotenoid content and color determination in the UAE oil extracts. UAE vegetable oil extracts enriched with carotenoids from papaya by-products could be useful to formulate new food ingredients based on emulsions with interesting potential health benefits.     

Antioxidant,Anti-Inflammatory,and Inhibition of Acetylcholinesterase Potentials of Cassia timoriensis DC. Flowers

Despite being widely used traditionally as a general tonic, especially in South East Asia, scientific research on Cassia timoriensis, remains scarce. In this study, the aim was to evaluate the in vitro activities for acetylcholinesterase (AChE) inhibitory potential, radical scavenging ability, and the anti-inflammatory properties of different extracts of C. timoriensis flowers using Ellman’s assay, a DPPH assay, and an albumin denaturation assay, respectively. With the exception of the acetylcholinesterase activity, to the best of our knowledge, these activities were reported for the first time for C. timoriensis flowers. The phytochemical analysis confirmed the existence of tannins, flavonoids, saponins, terpenoids, and steroids in the C. timoriensis flower extracts. The ethyl acetate extract possessed the highest phenolic and flavonoid contents (527.43 ± 5.83 mg GAE/g DW and 851.83 ± 10.08 mg QE/g DW, respectively) as compared to the other extracts. In addition, the ethyl acetate and methanol extracts exhibited the highest antioxidant (IC₅₀ 20.12 ± 0.12 and 34.48 ± 0.07 µg/mL, respectively), anti-inflammatory (92.50 ± 1.38 and 92.22 ± 1.09, respectively), and anti-AChE (IC₅₀ 6.91 ± 0.38 and 6.40 ± 0.27 µg/mL, respectively) activities. These results suggest that ethyl acetate and methanol extracts may contain bioactive compounds that can control neurodegenerative disorders, including Alzheimer’s disease, through high antioxidant, anti-inflammatory, and anti-AChE activities.     

Caralluma tuberculata N.E.Br Manifests Extraction Medium Reliant Disparity in Phytochemical and Pharmacological Analysis

Solubility of phytoconstituents depends on the polarity of the extraction medium used, which might result in the different pharmacological responses of extracts. In line with this, ethnomedicinally important food plant (i.e., Caralluma tuberculata extracts) have been made in fourteen distinct solvent systems that were then analyzed phytochemically via total phenolic amount estimation, total flavonoid amount estimation, and HPLC detection and quantification of the selected polyphenols. Test extracts were then subjected to a battery of in vitro assays i.e., antioxidants (DDPH scavenging, antioxidant capacity, and reducing power estimation), antimicrobial (antibacterial, antifungal, and antileishmanial), cytotoxic (brine shrimps, THP-1 human leukemia cell lines and normal lymphocytes), and protein kinase inhibition assays. Maximum phenolic and flavonoid contents were computed in distilled water–acetone and acetone extracts (i.e., 16 ± 1 μg/mg extract and 8 ± 0.4/mg extract, respectively). HPLC-DAD quantified rutin (0.58 µg/mg extract) and gallic acid (0.4 µg/mg extract) in methanol–ethyl acetate and methanol extracts, respectively. Water–acetone extract exhibited the highest DPPH scavenging of 36 ± 1%. Total reducing potential of 76.0 ± 1 μg/mg extract was shown by ethanol chloroform while maximum total antioxidant capacity was depicted by the acetone extract (92.21 ± 0.70 μg/mg extract). Maximal antifungal effect against Mucor sp., antileishmanial, brine shrimp cytotoxicity, THP-1 cell line cytotoxicity, and protein kinase inhibitory activities were shown by ethyl acetate-methanol (MIC: 50 µg/disc), n-hexane (IC₅₀: 120.8 ± 3.7 µg/mL), ethyl acetate (LD₅₀: 29.94 ± 1.6 µg/mL), distilled water–acetone (IC₅₀: 118 ± 3.4 µg/mL) and methanol–chloroform (ZOI: 19 ± 1 mm) extracts, respectively. Our findings show the dependency of phytochemicals and bioactivities on the polarity of the extraction solvent and our preliminary screening suggests the C. tuberculata extract formulations to be tested and used in different ailments, however, detailed studies remain necessary for corroboration with our results.     

Identification of Phenolic Compounds by LC-MS/MS and Evaluation of Bioactive Properties of Two Edible Halophytes: Limonium effusum and L. sinuatum

This work aimed to evaluate the phenolic content and in vitro antioxidant, antimicrobial and enzyme inhibitory activities of the methanol extracts and their fractions of two edible halophytic Limonium species, L. effusum (LE) and L. sinuatum (LS). The total phenolic content resulted about two-fold higher in the ethyl acetate fraction of LE (522.82 ± 5.67 mg GAE/g extract) than in that of LS (274.87 ± 1.87 mg GAE/g extract). LC-MS/MS analysis indicated that tannic acid was the most abundant phenolic acid in both species (71,439.56 ± 3643.3 µg/g extract in LE and 105,453.5 ± 5328.1 µg/g extract in LS), whereas hyperoside was the most abundant flavonoid (14,006.90 ± 686.1 µg/g extract in LE and 1708.51 ± 83.6 µg/g extract in LS). The antioxidant capacity was evaluated by DPPH and TAC assays, and the stronger antioxidant activity in ethyl acetate fractions was highlighted. Both species were more active against Gram-positive bacteria than Gram negatives and showed considerable growth inhibitions against tested fungi. Interestingly, selective acetylcholinesterase (AChE) activity was observed with LE and LS. Particularly, the water fraction of LS strongly inhibited AChE (IC₅₀ = 0.199 ± 0.009 µg/mL). The ethyl acetate fractions of LE and LS, as well as the n-hexane fraction of LE, exhibited significant antityrosinase activity (IC₅₀ = 245.56 ± 3.6, 295.18 ± 10.57 and 148.27 ± 3.33 µg/mL, respectively). The ethyl acetate fraction and methanol extract of LS also significantly inhibited pancreatic lipase (IC₅₀ = 83.76 ± 4.19 and 162.2 ± 7.29 µg/mL, respectively). Taken together, these findings warrant further investigations to assess the potential of LE and LS as a bioactive source that can be exploited in pharmaceutical, cosmetics and food industries.     

In Vitro Alpha-Glucosidase Inhibitory Activity and the Isolation of Luteolin from the Flower of Gymnanthemum amygdalinum (Delile) Sch. Bip ex Walp.

Diabetes mellitus is a major health issue that has posed a significant challenge over the years. Gymnanthemum amygdalinum is a well-known plant that can be potentially used to treat this disease. Therefore, this study aimed to evaluate the inhibitory effect of its root, stem bark, leaves, and flower extracts on alpha-glucosidase using an in vitro inhibition assay to isolate the bioactive compounds and determine their levels in the samples. The air-dried plant parts were extracted by maceration using methanol. The results showed that the flower extract had the greatest inhibitory effect (IC50 47.29 ± 1.12 µg/mL), followed by the leaves, roots, and stem bark. The methanolic flower extract was further fractionated with different solvents, and the ethyl acetate fraction showed the strongest activity (IC50 19.24 ± 0.12 µg/mL). Meanwhile, acarbose was used as a positive control (IC50 73.36 ± 3.05 µg/mL). Characterization based on UV, 1H-, and 13C-NMR established that the ethyl acetate fraction yielded two flavonoid compounds, namely, luteolin and 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-methoxy-4H-chromen-4-on, which had IC50 values of 6.53 ± 0.16 µg/mL and 39.95 ± 1.59 µg/mL, respectively. The luteolin levels in the crude drug, methanolic extract, and ethyl acetate fraction were 3.4 ± 0.2 mg (0.3%), 32.4 ± 0.8 mg (3.2%), and 68.9 ± 3.4 mg (6.9%) per 1 g samples, respectively. These results indicated that the G. amygdalinum flower extract exerted potent inhibitory alpha-glucosidase activity.     

Optimization of Ultrasound-Assisted Extraction of Phenolics from Sideritis raeseri Using Response Surface Methodology

In this study we define the optimal conditions for ultrasound-assisted extraction of bioactive polyphenols from S. raeseri aerial parts using response surface methodology. The influence of ethanol concentration (10–90%), extraction temperature (20–80 °C), extraction time (10–60 min), and solid-to-solvent ratio (1:10–1:50) on total phenolic content as well as on content of individual flavonoids, and hypolaetin and isoscutellarein derivatives was studied. For the experimental design, a central composite design was chosen. In the obtained extracts, the following ranges of targeted compounds were detected: total phenol from 19.32 to 47.23 mg GAE/g dw, HYP from 1.05 to 11.46 mg/g dw, ISC 1 from 0.68 to 10.68 mg/g dw, and ISC 2 from 0.74 to 15.56 mg/g dw. The optimal extraction conditions were set as: ethanol concentration of 65%, extraction time of 50 min, extraction temperature of 63 °C, and solid-to-solvent ratio of 1:40. Contents of TP, HYP, ISC 1, and ISC 2 in optimal extracts were 47.11 mg GAE/g dw, 11.73 mg/g dw, 9.54 mg/g dw, and 15.40 mg/g dw, respectively. Experimentally set values were in good agreement with those predicted by the response surface methodology model, indicating suitability of the used model, as well as the success of response surface methodology in optimizing the conditions of the extraction.     

In-Vitro Screenings for Biological and Antioxidant Activities of Water Extract from Theobroma cacao L. Pod Husk: Potential Utilization in Foods

Increasing production of cocoa (Theobroma cacao L.) leads to a higher environmental burden due to its solid waste generation. Cocoa pod husk, one of the major solid wastes of cocoa production, contains rich bioactive compounds unveiling its valorization potential. With that in mind, our research aimed to explore the biological and antioxidant activities of aqueous extracts from cocoa pod husks. In this present work, cocoa pod husk was extracted using water and subsequentially partitioned using n-hexane, ethyl acetate, and methanol. The antimicrobial investigation revealed that the ethyl acetate solubles were active against the Staphylococcus aureus, Escherichia coli, and Candida albicans, where at a 20% w/v concentration, the inhibition diameters were 6.62 ± 0.10, 6.52 ± 0.02, and 11.72 ± 0.36 mm, respectively. The extracts were found non-toxic proven by brine shrimp lethality tests against Artemia salina with LC₅₀ scores ranging from 74.1 to 19,054.6 μg/mL. The total phenolic content and total flavonoid content were obtained in the range of 47.44 to 570.44 mg/g GAE and 1.96 to 4.34 mg/g QE, respectively. Antioxidant activities of the obtained extracts were revealed by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay with EC₅₀ reached as low as 9.61 μg/mL by the ethyl acetate soluble. Phytochemical screening based on gas chromatography—mass spectroscopy analysis on the sample with the highest antioxidant activities revealed the dominant presence of three phytosterols, namely gamma-sitosterol, stigmasterol, and campesterol.     

Identification of Carotenoids in Hairless Canary Seed and the Effect of Baking on Their Composition in Bread and Muffin Products

Carotenoids are essential components in the human diet due to their positive functions in ocular and cognitive health. This study investigated composition of carotenoids in hairless canary seed (HCS) as a novel food and the effect of baking on carotenoids in bread and muffin made from HCS, wheat and corn. Three bread formulations made from wheat and HCS blends were evaluated and compared with control wheat bread. In addition, three low-fat muffin recipes prepared from HCS alone or in blends with corn were assessed. The fate of carotenoid compounds in breads and muffins was monitored after dry mixing, dough/batter formation and oven baking. Carotenoids in products were quantified using UPLC and their identification was confirmed based on LC-MS/MS. Hairless canary seed and corn were fairly rich in carotenoids with a total content of 7.6 and 12.9 µg/g, respectively, compared with wheat (1.3 µg/g). Nineteen carotenoid compounds were identified, with all-trans lutein being the principal carotenoid in HCS followed by lutein 3-O-linoleate, lutein 3-O-oleate and lutein di-linoleate. There were significant reductions in carotenoids in muffin and bread products. It appears that batter or dough preparation causes more reductions in carotenoids than oven baking, probably due to enzymatic oxidation and degradation. Muffin-making resulted in lower lutein reductions compared with the bread-making process. The results suggest that muffins made from hairless canary seed alone or in blends with corn could boost the daily intake of lutein and/or zeaxanthin.     

Simultaneous extraction and purification of fucoxanthin from Tisochrysis lutea microalgae using compressed fluids

The marine microalga Tisochrysis lutea, a Haptophyta with a thin cell wall and currently used mainly in aquaculture is a potential source of several bioactive compounds of interest such as carotenoids. In the present study, the simultaneous extraction and purification of fucoxanthin, the main carotenoid from T. lutea, was optimized using pressurized fluid extraction followed by in‐cell purification. An experimental design was employed to maximize carotenoids’ extraction; the experimental factors chosen were: (i) percentage of ethanol/ethyl acetate (0–100 %), (ii) temperature (40–150°C), and (iii) number of static extraction cycles (1–3). The maximum carotenoids’ recovery, mainly fucoxanthin, was obtained with pure ethyl acetate at 40°C using one extraction cycle, achieving values of 132.8 mg of carotenoids per gram of extract. Once the optimum extraction conditions were confirmed, in‐cell purification strategies using different adsorbents were developed to obtain fucoxanthin‐enriched extracts. Activated charcoal showed potential retention of chlorophylls allowing an effective purification of fucoxanthin in the obtained extracts. Chemical characterization of extracts was carried out by reversed‐phase high‐performance liquid chromatography with diode array detection. Therefore, a selective fractionation of high value compounds was achieved using the proposed green downstream platform based on the use of compressed fluids.     

Comparative Evaluation of the Phytochemical Profiles and Antioxidant Potentials of Olive Leaves from 32 Cultivars Grown in China

Olives (Olea europaea L.) are a significant part of the agroindustry in China. Olive leaves, the most abundant by-products of the olive and olive oil industry, contain bioactive compounds that are beneficial to human health. The purpose of this study was to evaluate the phytochemical profiles and antioxidant capacities of olive leaves from 32 cultivars grown in China. A total of 32 phytochemical compounds were identified using high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry, including 17 flavonoids, five iridoids, two hydroxycinnamic acids, six triterpenic acids, one simple phenol, and one coumarin. Specifically, olive leaves were found to be excellent sources of flavonoids (4.92–18.29 mg/g dw), iridoids (5.75–33.73 mg/g dw), and triterpenic acids (15.72–35.75 mg/g dw), and considerable variations in phytochemical content were detected among the different cultivars. All tested cultivars were classified into three categories according to their oil contents for further comparative phytochemicals assessment. Principal component analysis indicated that the investigated olive cultivars could be distinguished based upon their phytochemical profiles and antioxidant capacities. The olive leaves obtained from the low-oil-content (<16%) cultivars exhibited higher levels of glycosylated flavonoids and iridoids, while those obtained from high-oil-content (>20%) cultivars contained mainly triterpenic acids in their compositions. Correspondingly, the low-oil-content cultivars (OL3, Frantoio selection and OL14, Huaou 5) exhibited the highest ABTS antioxidant activities (758.01 ± 16.54 and 710.64 ± 14.58 mg TE/g dw, respectively), and OL9 (Olea europaea subsp. Cuspidata isolate Yunnan) and OL3 exhibited the highest ferric reducing/antioxidant power assay values (1228.29 ± 23.95 mg TE/g dw and 1099.99 ± 14.30 mg TE/g dw, respectively). The results from this study may be beneficial to the comprehensive evaluation and utilization of bioactive compounds in olive leaves.     

HPTLC and FTIR Fingerprinting of Olive Leaves Extracts and ATR-FTIR Characterisation of Major Flavonoids and Polyphenolics

The aim of this study was to analyse the effect of spontaneous microbial maceration on the release and extraction of the flavonoids and phenolics from olive leaves. Bioprofiling based on thin-layer chromatography effect-directed detection followed by ATR-FTIR spectroscopy proved to be a reliable and convenient method for simultaneous comparison of the extracts. Results show that fermentation significantly enhances the extraction of phenolic compounds and flavonoids. The polyphenolic content was increased from 6.7 µg GAE (gallic acid equivalents) to 25.5 µg GAE, antioxidants from 10.3 µg GAE to 25.3 µg GAE, and flavonoid content from 42 µg RE (rutin equivalents) to 238 µg RE per 20 µL of extract. Increased antioxidant activity of fermented ethyl acetate extracts was attributed to the higher concentration of extracted flavonoids and phenolic terpenoids, while increased antioxidant activity in fermented ethanol extract was due to increased extraction of flavonoids as extraction of phenolic compounds was not improved. Lactic acid that is released during fermentation and glycine present in the olive leaves form a natural deep eutectic solvent (NADES) with significantly increased solubility for flavonoids.     

Valorization of Phenolic and Carotenoid Compounds of Sechium edule (Jacq. Swartz) Leaves: Comparison between Conventional,Ultrasound- and Microwave-Assisted Extraction Approaches

Chayote leaves are known for culinary and traditional medicine applications. This work intended to recover carotenoids and phenolic compounds from chayote leaves using the ultrasound-assisted extraction (UAE). A Box–Behnken design was employed to investigate the impact of extraction time, temperature, and ultrasonic power on the recovery of total carotenoids, total phenolic compounds, and antioxidant activities. For comparative purposes, chayote leaf extracts were prepared by maceration (ME) and microwave-assisted extraction (MAE), using the same time and temperature conditions optimized by UAE. Extraction at 50 °C and 170 Watts for 30 min provided the optimal UAE conditions. UAE showed better extraction efficacy than ME and MAE. The HPLC analysis of the extracts showed that the xanthophyll class was the main class of carotenoids, which constituted 42–85% of the total carotenoid content, followed by β-carotene and tocopherol. Moreover, 26 compounds, classified as phenolic acids, flavonols, flavonoids and other polar compounds, were identified in the chayote leaf extracts. Flavonols accounted for 55% of the total compounds quantified (the major compound was myricetin) and phenolic acids represented around 35%, mostly represented by ferulic acid, chlorogenic acid and (+)-catechin. This study revealed the potential of UAE as an effective green extraction technique to recover bioactive compounds from chayote leaves, for food, and for pharmaceutical and cosmetic applications.     

An Insight into Kiwiberry Leaf Valorization: Phenolic Composition,Bioactivity and Health Benefits

During kiwiberry production, different by-products are generated, including leaves that are removed to increase the fruit’s solar exposure. The aim of this work was to extract bioactive compounds from kiwiberry leaf by employing microwave-assisted extraction (MAE). Compatible food solvents (water and ethanol) were employed. The alcoholic extract contained the highest phenolic and flavonoid contents (629.48 mg of gallic acid equivalents (GAE) per gram of plant material on dry weight (dw) (GAE/g dw) and 136.81 mg of catechin equivalents per gram of plant material on dw (CAE/g dw), respectively). Oppositely, the hydroalcoholic extract achieved the highest antioxidant activity and scavenging activity against reactive oxygen and nitrogen species (IC₅₀ = 29.10 μg/mL for O₂^•−, IC₅₀ = 1.87 μg/mL for HOCl and IC₅₀ = 1.18 μg/mL for ^•NO). The phenolic profile showed the presence of caffeoylquinic acids, proanthocyanidin, and quercetin in all samples. However, caffeoylquinic acids and quercetin were detected in higher amounts in the alcoholic extract, while proanthocyanidins were prevalent in the hydroalcoholic extract. No adverse effects were observed on Caco-2 viability, while the highest concentration (1000 µg/mL) of hydroalcoholic and alcoholic extracts conducted to a decrease of HT29-MTX viability. These results highlight the MAE potentialities to extract bioactive compounds from kiwiberry leaf.     

Novel Processes for the Extraction of Phenolic Compounds from Olive Pomace and Their Protection by Encapsulation

Olive pomace, the solid by-product derived from olive oil production consists of a high concentration of bioactive compounds with antioxidant activity, such as phenolic compounds, and their recovery by applying innovative techniques is a great opportunity and challenge for the olive oil industry. This study aimed to point out a new approach for the integrated valorization of olive pomace by extracting the phenolic compounds and protecting them by encapsulation or incorporation in nanoemulsions. Innovative assisted extraction methods were evaluated such as microwave (MAE), homogenization (HAE), ultrasound (UAE), and high hydrostatic pressure (HHPAE) using various solvent systems including ethanol, methanol, and natural deep eutectic solvents (NADESs). The best extraction efficiency of phenolic compounds was achieved by using NADES as extraction solvent and in particular the mixture choline chloride-caffeic acid (CCA) and choline chloride-lactic acid (CLA); by HAE at 60 °C/12,000 rpm and UAE at 60 °C, the total phenolic content (TPC) of extracts was 34.08 mg gallic acid (GA)/g dw and 20.14 mg GA/g dw for CCA, and by MAE at 60 °C and HHPAE at 600 MPa/10 min, the TPC was 29.57 mg GA/g dw and 25.96 mg GA/g dw for CLA. HAE proved to be the best method for the extraction of phenolic compounds from olive pomace. Microencapsulation and nanoemulsion formulations were also reviewed for the protection of the phenolic compounds extracted from olive pomace. Both encapsulation techniques exhibited satisfactory results in terms of encapsulation stability. Thus, they can be proposed as an excellent technique to incorporate phenolic compounds into food products in order to enhance both their antioxidative stability and nutritional value.     

HPLC-DAD Based Polyphenolic Profiling and Evaluation of Pharmacological Attributes of Putranjiva roxburghii Wall.

The current study was intended to explore the phytochemical profiling and therapeutic activities of Putranjiva roxburghii Wall. Crude extracts of different plant parts were subjected to the determination of antioxidant, antimicrobial, antidiabetic, cytotoxic, and protein kinase inhibitory potential by using solvents of varying polarity ranges. Maximum phenolic content was notified in distilled water extracts of the stem (DW-S) and leaf (DW-L) while the highest flavonoid content was obtained in ethyl acetate leaf (EA-L) extract. HPLC-DAD analysis confirmed the presence of various polyphenols, quantified in the range of 0.02 ± 0.36 to 2.05 ± 0.18 μg/mg extract. Maximum DPPH scavenging activity was expressed by methanolic extract of the stem (MeOH-S). The highest antioxidant capacity and reducing power was shown by MeOH-S and leaf methanolic extract (MeOH-L), respectively. Proficient antibacterial activity was shown by EA-L extract against Bacillus subtilis and Escherichia coli. Remarkable α-amylase and α-glucosidase inhibition potential was expressed by ethyl acetate fruit (EA-F) and n-Hexane leaf (nH-L) extracts, respectively. In case of brine shrimp lethality assay, 41.67% of the extracts (LC₅₀ < 50 µg/mL) were considered as extremely cytotoxic. The test extracts also showed mild antifungal and protein kinase inhibition activities. The present study explores the therapeutic potential of P. roxburghii and calls for subsequent studies to isolate new bioactive leads through bioactivity-guided isolation.     

The Identification and Quantitative Analysis of Unusual Keto-Carotenoids in Ripe Fruits of Maclura tricuspidate and Its Potential as a Valuable Source of Cryptocapsin

Ripe fruits of Maclura tricuspidata (MT) are used as food material and a natural colorant in Korea. Although MT fruits have a deep red color due to carotenoid-like pigments, their chemical nature has not been explored in detail so far. The present study aimed at elucidating the chemical structures and composition of carotenoids in MT fruits and changes at different maturity stages. Two carotenoids from saponified MT fruit extract were isolated using repeated silica gel column chromatography. Based on interpretations of spectroscopic data, these compounds were determined as keto-carotenoids, i.e., capsanthin (3,3′-dihydroxy-β,κ-caroten-6′-one) and cryptocapsin (3′-hydroxy-β,κ-caroten-6’-one), and the contents of individual carotenoids were quantified with HPLC based on calibration curves obtained from authentic standards. The contents of capsanthin and cryptocapsin in the sample of saponified MT fruits were 57.65 ± 1.97 µg/g and 171.66 ± 4.85 μg/g as dry weight base (dw). The majority of these keto-carotenoids in the MT fruits were present in esterified forms with lauric, myristic or palmitic acid rather than in their free forms. The results also showed that esterification of these compounds occurred starting from early stage (yellow-brownish stage) of maturation. Considering the high cryptocapsin content, MT fruits can be applied as a potentially valuable source of cryptocapsin for food and medicinal application as well as a source of provitamin A.     

Supercritical CO2 Extraction of High-Added Value Compounds from Chlorella vulgaris: Experimental Design,Modelling and Optimization

Microalgae are well-known for their high-added value compounds and their recovery is currently of great interest. The aim of this work is the recovery of such components from Chlorella vulgaris through supercritical fluid extraction (SFE) with CO₂. The effect of the extraction temperature (40–60 °C), pressure (110–250 bar), and solvent flow rate (20–40 g/min) was tested on yield, the extract’s antioxidant activity, and the phenolic, chlorophyll and carotenoid content. Thus, data analysis indicated that the yield was mainly affected by temperature, carotenoids by pressure, while the extract’s phenolics and antioxidant activity were affected by the synergy of temperature and pressure. Moreover, SFE’s kinetic study was performed and experimental data were correlated using Sovová’s mass transfer-based model. SFE optimization (60 °C, 250 bar, 40 g/min) led to 3.37% w/w yield, 44.35 mg_extr/mg_DPPH antioxidant activity (IC50), 18.29 mg_GA/g_extr total phenolic content, 35.55, 21.14 and 10.00 mg/g_extr total chlorophyll, carotenoid and selected carotenoid content (astaxanthin, lutein and β-carotene), respectively. A comparison of SFE with conventional aq. ethanol (90% v/v) extraction proved SFE’s superiority regarding extraction duration, carotenoids, antioxidant activity and organoleptic characteristics of color and odor despite the lower yield. Finally, cosolvent addition (ethanol 10% w/w) at optimum SFE conditions improved the extract’s antioxidant activity (19.46%) as well as yield (101.81%).     

Impact of Peels Extracts from an Italian Ancient Tomato Variety Grown under Drought Stress Conditions on Vascular Related Dysfunction

Background: Tomato by-products contain a great variety of biologically active substances and represent a significant source of natural antioxidant supplements of the human diet. The aim of the work was to compare the antioxidant properties of a by-product from an ancient Tuscan tomato variety, Rosso di Pitigliano (RED), obtained by growing plants in normal conditions (-Ctr) or in drought stress conditions (-Ds) for their beneficial effects on vascular related dysfunction. Methods: The antioxidant activity and total polyphenol content (TPC) were measured. The identification of bioactive compounds of tomato peel was performed by HPLC. HUVEC were pre-treated with different TPC of RED-Ctr or RED-Ds, then stressed with H₂O₂. Cell viability, ROS production and CAT, SOD and GPx activities were evaluated. Permeation of antioxidant molecules contained in RED across excised rat intestine was also studied. Results: RED-Ds tomato peel extract possessed higher TPC than compared to RED-Ctr (361.32 ± 7.204 mg vs. 152.46 ± 1.568 mg GAE/100 g fresh weight). All extracts were non-cytotoxic. Two hour pre-treatment with 5 µg GAE/mL from RED-Ctr or RED-Ds showed protection from H₂O₂-induced oxidative stress and significantly reduced ROS production raising SOD and CAT activity (* p < 0.05 and ** p < 0.005 vs. H₂O₂, respectively). The permeation of antioxidant molecules contained in RED-Ctr or RED-Ds across excised rat intestine was high with non-significant difference between the two RED types (41.9 ± 9.6% vs. 26.6 ± 7.8%). Conclusions: RED-Ds tomato peel extract represents a good source of bioactive molecules, which protects HUVECs from oxidative stress at low concentration.     

Selective Extraction of Sinapic Acid Derivatives from Mustard Seed Meal by Acting on pH: Toward a High Antioxidant Activity Rich Extract

The aim of this paper is to study the effect of the pH on the extraction of sinapic acid and its derivatives from mustard seed meal. Solutions of acidic pH (pH 2), basic pH (pH 12) and distilled water (uncontrolled pH ~ 4.5) were tested at different percentages of ethanol. The maximum extraction yield for sinapic acid (13.22 µmol/g of dry matter (DM)) was obtained with a buffered aqueous solution at pH 12. For ethyl sinapate, the maximum extraction yield reached 9.81 µmol/g _DM with 70% ethanol/buffered aqueous solution at pH 12. The maximum extraction yield of sinapine (15.73 µmol/g _DM) was achieved with 70% ethanol/buffered aqueous solution at pH 2. The antioxidant activity of each extract was assessed by DPPH assay; the results indicated that the extracts obtained at pH 12 and at low ethanol percentages (<50%) exhibit a higher antioxidant activity than extracts obtained at acidic conditions. Maximum antioxidant activity was reached at pH 12 with buffer solution (11.37 mg of Trolox Equivalent/g _DM), which confirms that sinapic acid-rich fractions exhibit a higher antioxidant activity. Thus, to obtain rich antioxidant extracts, it is suggested to promote the presence of sinapic acid in the extracts.     

Optimization and development of a high‐performance liquid chromatography method for the simultaneous determination of vitamin E and carotenoids in tomato fruits

A simple and reliable high‐performance liquid chromatography method was developed and validated for the simultaneous determination of lipophilic antioxidants in tomato fruits using C₃₀ column operated at 15°C and a gradient mobile phase based on acetonitrile/methanol/dichloromethane in a total run time of 30 min. Diode array and fluorescence detectors were used respectively for the detection of carotenoids (lutein, zeaxanthin, cryptoxanthin, lycopene, and β‐carotene) and vitamin E analogs (α‐, β‐, γ‐, and δ‐tocopherols, and tocotrienols). The best extraction yield of analytes in tomato fruits was achieved by employing ethyl acetate/hexane (1:1, v/v) after several treatments with various solvents. In addition, low extraction yields were obtained for carotenoids compared to tocopherols by adopting solid‐phase extraction as a second clean‐up step. The method was validated on the basis of recovery, precision, linearity, and limit of detection and quantification using spiked tomato samples. The method was applied to cherry and medium‐sized tomato fruits. Lycopene was found to be present in largest amount in tomato pulp, followed by β‐carotene and lutein. Due to its simplicity, rapidity, and efficiency, the method is suitable for routine analysis of lipophilic antioxidants in tomato fruits, and may also be applied to other vegetables of similar phytochemical profiles.