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Antioxidant and Cytotoxic Activities of Carob Tree Fruit Pulps Are Strongly Influenced by Gender and Cultivar
Abstract
Extracts from fruit pulps of six female cultivars and two hermaphrodite Portuguese carob trees [(Ceratonia siliqua L., Fabaceae)] exhibited strong antioxidant activity and were rich in phenolic compounds. The extracts decreased the viability of different human cancer cell lines on a dose- and time-dependent manner. Gender and cultivar significantly influenced the chemical content and the biological activities of the extracts. Extracts from hermaphrodite trees had a higher content of phenolic compounds, and exhibited higher antioxidant and cytotoxic activities. Among females, cv. Aida had the highest radical scavenging activity and total content of phenolics, Mulata the highest capacity to inhibit lipid oxidation and Gasparinha the strongest cytotoxic activity on HeLa cells. The decrease in cell viability was associated with apoptosis on HeLa and MDA-MB-231 lines. (+)-Catechin and gallic acid (GA) were the main compounds identified in the extracts, and GA contributed to the antioxidant activity. Our results show that the antioxidant and cytotoxic activities of carob tree fruit pulps are strongly influenced by gender and cultivar, and provide new knowledge about the advantages of hermaphrodite trees over female cultivars, namely, as a source of compounds with biological interest, which may represent an increase of their agronomic interest.
... According to Purohit and Gogate [10], the use of a lower solid to solvent ratio than the optimum value leads to an increase in the solvent consumption and higher solid to solvent ratios than the optimum value will result in an incomplete extraction. Using a solid to solvent ratio of 1:10, the yield of the extraction was higher than when using a 1:5 solid to solvent ratio, and this could be explained by the excessive amount of microalgal material (1:5 ratio), which caused the increase in viscosity, and thus inhibited the diffusion of compounds through the extraction medium [11]. In addition, it contributed to ultrasound wave attenuation, leaving the restricted zone located near the ultrasound probe as the active part [12]. ...
... The extraction process usually becomes more energy efficient at a higher algal biomass concentration because it is more effective in its wave contact with solid matter, such as microalgae cells, as for all released components. However, in some cases, when an excessive amount of algal biomass is present, the diffusion of compounds towards the extraction medium becomes difficult due to an increase in viscosity, as previously mentioned [11]. ...
... The solid to solvent ratio also affected the extraction efficiency: the carbohydrate content was higher in the extracts than in the biomass, which was more evident when the solid to solvent ratio was 1:5, either in continuous or pulse sonication modes. The same trend was reported by Zhao and coworkers [23]; however, it was the opposite of what we expected because the excessive amount of microalgal material (1:5 ratio) usually inhibited the diffusion of compounds through a more viscous extraction medium [11]. The sonication mode alone had no influence on the carbohydrate's extraction yield. ...
(1) Background: Microalgae are promising feedstock for obtaining valuable bioactive compounds. To facilitate the release of these important biomolecules from microalgae, effective cell disruption is usually necessary, where the use of ultrasound has achieved considerable popularity as an alternative to conventional methods. (2) Methods: This paper aims to evaluate the use of ultrasound technology in water medium as a green technology to recover high added-value compounds from Dunaliella salina and improve its sensory profile towards a high level of incorporation into novel food products. (3) Results: Among the variables, the solid concentration and extraction time have the most significant impact on the process. For the extraction of protein, or fat, the most influential factor is the extraction time. Total polyphenols are only significantly affected by the extraction time. The antioxidant capacity is strongly affected by the solid to liquid ratio and, to a small extent, by the extraction time. Ultrasound-assisted extraction improves the overall odor/aroma of D. salina with good acceptability by the panelists. (4) Conclusions: The application of ultrasonic-assisted extraction demonstrates a positive overall effect on enhancing the sensory profile, particularly the odor of microalgal biomass, while the bioactive properties are preserved. Notably, the intense sea/fish odors are reduced, while earthy and citrus notes become more prominent, resulting in an improved overall sensory profile score. This is the first time, to our knowledge, that this innovative, green, and efficient technology has been used to upgrade the aroma profile of microalgae.
... The present study focuses on the comparative analysis of carob extracts obtained from two distinct C. siliqua cultivars encountered in different geographical locations on the island of Crete. It has been shown previously that gender and cultivar significantly influence the chemical content and the biological activities of carob extracts with Portuguese origin [33]. This also seems to be the case with the two Cretan cultivars of the present study, at least concerning the chemical content; only 11 out of 27 essential oil compounds (41%) were found to be common in the pods of the two cultivars, while also the fatty acid and the carbohydrate composition varied considerably (Tables 2-4). ...
... We did not locate any other studies in the literature regarding carob extracts' effects on human skin fibroblasts; however, liposome encapsulated C. siliqua pod extracts tested on 3T3 embryonic mouse fibroblasts, and immortalized human keratinocytes (HaCaT) have been reported to be noncytotoxic [37], in accordance with our findings. There are many studies in the literature regarding carob extracts' cytotoxicity towards a variety of different cancer cell lines [33,[38][39][40][41][42][43][44][45][46][47][48][49][50][51]. Cytotoxic activity has been mainly found with polar extracts and is believed to be a result of synergy between many compounds contained in carob [7,44]. ...
... In general, the IC 50 values reported in the literature for methanolic carob pod extracts vary from around 100-200 µg/mL [61] to around 10 mg/mL [33], and this may be due to the different geographical origins, the cultivar, the gender, or the ripening stage [62]. Regarding the two Cretan cultivars presented here, the reduction in DPPH absorbance for CSE1 and CSH1 (Figure 2A) was not sufficient to calculate an IC 50 (the highest concentration tested being 2 mg/mL); however, IC 50 values of 1177 ± 240, 17 ± 1, and 79 ± 22 µg/mL were obtained from fractions CSE1.1, CSE1.2, and CSE1.3, respectively ( Figure 6A). ...
Ceratonia siliqua L., commonly known as the carob tree, appears in most Mediterranean countries, often cultivated for the collection of its fruits to be used as food for humans and animals. This study was aimed at the phytochemical characterization of two common Cretan C. siliqua cultivars and the biological evaluation of deseeded pod and seed extracts regarding their putative use in cosmetics. Gas and liquid chromatographic techniques were used to assess their essential oil, fatty acid, and carbohydrate profiles. Cell-free assays, including free-radical scavenging; the inhibition of tyrosinase and collagenase; the blocking of advanced glycation end product (AGE) formation; along with assays in human skin fibroblast cultures, i.e., reactive oxygen species suppression, glutathione stimulation, and protection from oxidative stress and from ultraviolet (UVB) radiation, were also used. Extracts from both cultivars were found to possess antioxidant capacity, tyrosinase- and collagenase-inhibitory activities, an ability to block glucose-induced AGEs, and in certain cases, UVB absorbance and photoprotective activities. Seed extracts were in general more active, while the use of 30% aqueous methanol seemed to be more efficient than n-hexane for extraction. Serial partition of the most active extracts resulted in fractions with enriched biological activities. These properties make Cretan carob extracts and their fractions suitable candidates for use in cosmetics.
... Carob tree (Ceratonia siliqua L.), an evergreen tree belonging to the Fabaceae family, is native to the Mediterranean area, but is also grown in some regions of the United States, Latin America, and Australia (Custodio et al., 2011;Ben Ayache et al., 2020). In 2020, Algeria participated by 6.6% in the world production of carob fruit, which is estimated at about 49,693 tons; with Portugal, Morocco, and Greece being the major producers of this fruit (FAO, 2022). ...
... Furthermore, the presence of biomolecules such us phenolic compounds, flavonoids, tannins, and anthocyanins in carob pods may have beneficial effects on human and animal health. Numerous studies reported that the extracts of carob exhibit an antioxidant, antidiarrheal, antibacterial, antifungal, anti-inflammatory, antidiabetic activities, and also hepatoprotective and antiproliferative effects (Custodio et al., 2011;Kotrotsios et al., 2012;Lakkab et al., 2018;Ben Ayache et al., 2020). Carob and carob byproducts have been studied previously as a local alternative feed and to prevent and treat diarrhea and as anthelmintic in geese (Sahle et al., 1992), broiler chicken (Ortiz et al., 2004), fish (Fadel et al., 2017), quail (Calislar and Kaplan, 2017) and sheep (Saratsi et al., 2020) diets. ...
... Phenolic compounds are non-nutrient but biologically active secondary plant metabolites, which can act as antioxidants (Vekiari et al., 2012). In our study, carob pulp powder contained significantly higher total phenolic content (29.37 § 1.21 mg GAE/g DM) compared to the range (17−19 mg GAE/g DM) reported by Custodio et al. (2011) and Kyriacou et al. (2021). The main constituents of carob polyphenols were found to be condensed tannins (Vekiari et al., 2012). ...
In this study, the influence of a partial substitution of corn with carob (Ceratonia siliqua L.) pulp powder on broiler performance, intestinal microbiota, carcass traits and meat quality, was investigated. Two dietary treatments were compared: the control group received a diet containing basically corn, soybean meal and wheat bran for all the breeding periods while the experimental group received the same starter diet as the control group and grower and finisher diets containing 3% and 7% of carob pulp powder, respectively. Growth performance (weight gain, feed intake and feed conversion ratio), carcass traits, lactic acid bacteria, and total coliforms were determined. Physicochemical parameters, proximate composition, fatty acid profiles, lipid oxidation index and sensory characteristics of broiler thigh and breast muscles were determined. Dietary supplementation with carob pulp powder did not show an effect (p>0.05) on growth performance and carcass traits except for slaughter yield and breast relative weight. An increase in sum lactic acid bacteria count and a decrease in total coliforms were observed in the experimental group. No effect (p>0.05) of dietary carob was observed on pH, cooking losses, ash, mineral, and protein contents in breast and thigh muscles from broilers. However, the experimental diet decreased (p<0.05) moisture and fat contents in thigh muscles but not in breast muscles. Both muscles from the experimental group of birds exhibited lower palmitic and oleic acids contents, and higher linoleic and linolenic acids levels than those from the control group. Whereas the polyunsaturated fatty acid contents in broiler breast and thigh meat was increased by dietary carob, the concentration of lipid oxidation products decreased in this group. The dietary intervention had no significant effect on the sensory profile of breast and thigh muscles. Thus, carob pulp powder can be applied to broiler diet at 7% as a nonconventional feed with no negative impact on growth performance and carcass traits and to increase the degree of unsaturation with no negative impact on oxidative stability or sensory traits.
... Microsatellites (SSRs) have been successfully used to assess carob genetic diversity (La Malfa et al., 2014;Viruel et al., 2018;Di Guardo et al., 2019). Additionally, a prerequisite for revitalizing the carob industry is the assessment of the variation for morphological and compositional traits (Barracosa et al., 2007;Custodio et al., 2011;Benchikh et al., 2014). However, for rain fed crops, such as carob, grown under marginal environments, the agro-environmental effect owing to spatial variation in edapho-climatic conditions would be expected to predominate over the genotypic effect (Blum, 2010). ...
... Gallic acid was the major phenolic compound and mallic acid the major organic acid ( Ayaz et al., 2007;Farag et al., 2019). Contrary to Portuguese carobs (Custodio et al., 2011), catechin appeared at very low concentration. Potassium was the major metal detected in the pulp accompanied by considerable concentration of calcium, phosphorus and magnesium (Oziyci et al., 2014;Goulas et al., 2016). ...
... Potassium was the major metal detected in the pulp accompanied by considerable concentration of calcium, phosphorus and magnesium (Oziyci et al., 2014;Goulas et al., 2016). Morphological and compositional traits of carobs can be affected by genotype (Barracosa et al., 2007), gender (Custodio et al., 2011), climatic conditions (Tous et al., 2013), harvest stage (Benchikh et al., 2014;Farag et al., 2019), soil conditions, and season (Correia et al., 2018). In the present study, genotypic effect was high for pod morphological traits, particularly for the seeds/pod weight ratio, pod width and thickness, as well as total sugars. ...
... Microsatellites (SSRs) have been successfully used to assess carob genetic diversity (La Malfa et al., 2014;Viruel et al., 2018;Di Guardo et al., 2019). Additionally, a prerequisite for revitalizing the carob industry is the assessment of the variation for morphological and compositional traits (Barracosa et al., 2007;Custodio et al., 2011;Benchikh et al., 2014). However, for rain fed crops, such as carob, grown under marginal environments, the agro-environmental effect owing to spatial variation in edapho-climatic conditions would be expected to predominate over the genotypic effect (Blum, 2010). ...
... Gallic acid was the major phenolic compound and mallic acid the major organic acid ( Ayaz et al., 2007;Farag et al., 2019). Contrary to Portuguese carobs (Custodio et al., 2011), catechin appeared at very low concentration. Potassium was the major metal detected in the pulp accompanied by considerable concentration of calcium, phosphorus and magnesium (Oziyci et al., 2014;Goulas et al., 2016). ...
... Potassium was the major metal detected in the pulp accompanied by considerable concentration of calcium, phosphorus and magnesium (Oziyci et al., 2014;Goulas et al., 2016). Morphological and compositional traits of carobs can be affected by genotype (Barracosa et al., 2007), gender (Custodio et al., 2011), climatic conditions (Tous et al., 2013), harvest stage (Benchikh et al., 2014;Farag et al., 2019), soil conditions, and season (Correia et al., 2018). In the present study, genotypic effect was high for pod morphological traits, particularly for the seeds/pod weight ratio, pod width and thickness, as well as total sugars. ...
Carob is a predominantly rainfed tree crop of high nutritive value and a long history of adaptation to the edaphoclimatic stress conditions of the Mediterranean. However, declining attention to the carob tree in recent decades has aggravated genetic erosion. The extant in situ germplasm varies both in terms of pod morphology and composition, reflecting the genetic and physiological divide chiefly among grafted and non-grafted material, and possibly the impact of variable agro-environments. Accordingly, the present study aimed to establish a systematic categorization of the genetic and phenotypic diversity encountered across carob germplasm identified in situ throughout Cyprus, a historical center of production and genetic diversity for the species. Linking pod morphology, primary and secondary metabolite profiles with genotyped source material originating in different agro-environments and crop seasons would provide a framework for interpreting (a) the interaction of these factors in configuring carob pod physicochemical constitution, and (b) the relative stability of phenotypic traits against environmental and seasonal variation. Microsatellite analysis discriminated 36 genotypes out of the 124 trees located in nine traditional agro-environmental zones and revealed low genetic diversity within the grafted germplasm. Two landraces were identified: "Tillyria," which is widespread and predominant, and "Kountourka," which is mainly localized to the northeastern peninsula of Karpasia. Morphological traits, such as seeds-to-pod weight ratio, pod width and thickness were principally under genetic control. Contrarily, compositional traits, particularly total phenolic content-including condensed tannins, in vitro antioxidant capacity and to a lesser extent gallic acid, organic acids and minerals were under agro-environmental control. Agro-environmental zone also modulated principally fructose and glucose; sucrose was modulated equally by genotype and agro-environment, while total sugars were under genetic control. Statistically significant differences between seasons were detected for all traits except for the seeds-to-pod weight ratio, pod length and width. Hierarchical cluster analysis corroborates that Cyprus may be divided into two major agro-environmental zones Frontiers in Plant Science | www.frontiersin.org 1 January 2021 | Volume 11 | Article 612376 Kyratzis et al. Carob Pod Morphology and Metabolite Profile modulating the compositional properties of the carob pulp. The present study provides a comprehensive insight into the extant carob genetic resources of Cyprus and advances our understanding of how genetic, agro-environmental and seasonal factors interact in shaping carob pod morphology and composition.
... The TPC reached a minimum concentration at RS5 (17.4 ± 2.0 mg GAE g −1 dw) and remained unaltered through RS6. Thus, the pulp of Cypriot carob genotypes at full commercial maturity had comparable TPC to the mean content previously reported for cultivars from Italy (16.0 ± 2.1 mg GAE g −1 dw) [5], Portugal (19.0 ± 3.0 mg GAE g −1 dw) [51] and Greece (18.0 ± 0.8 mg GAE g −1 dw) [17]. The TPC reported in different studies, however, can vary significantly, and variations can be traced to differences in the genetic material, ripeness and agroenvironmental conditions [2]. ...
... Thus, a significant reduction in condensed tannins can occur even after the carob fruit attains complete coloration. The levels of condensed tannins (46.45 ± 8.50 mg CE g −1 dw) present in the Cypriot carob genotypes at the late ripe stage (RS6) were much higher than those previously reported for Italian [5] and Portuguese carobs [51]. However, the variations among the carob genotypes in condensed tannins can be very high, as, for instance, between Portuguese cultivars Preta de Lagos (1.1 ± 0.1 mg CE g −1 ) and Costela (10.0 ± 0.3 mg CE g −1 [51]. ...
... The levels of condensed tannins (46.45 ± 8.50 mg CE g −1 dw) present in the Cypriot carob genotypes at the late ripe stage (RS6) were much higher than those previously reported for Italian [5] and Portuguese carobs [51]. However, the variations among the carob genotypes in condensed tannins can be very high, as, for instance, between Portuguese cultivars Preta de Lagos (1.1 ± 0.1 mg CE g −1 ) and Costela (10.0 ± 0.3 mg CE g −1 [51]. Significant variations may also derive from different sampling environments, extraction solvents of differing acidity and quantification methods showing distinct specificities [54]. ...
Six critical stages corresponding to major morphophysiological events in carob fruit ripening were defined, and changes in the primary and secondary metabolome and in vitro antioxidant capacity were examined in two genotypes collected at low (15 m) and high (510 m) altitudes from genetically identified and georeferenced trees. Soluble carbohydrates were analyzed by HPLC-RI, macro-minerals by ion chromatography coupled to conductivity detection and polyphenols by UHPLC-Q-Orbitrap-HRMS. spectroscopy facilitated assays for condensed tannins and in vitro free-radical scavenging capacity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric-reducing antioxidant power (FRAP). The fruit respiration rate and moisture content declined sharply during the transition from the breaker to green pedicel stage. Sugar accumulation spiked at the onset of fruit coloration and culminated at 498.7 ± 8.4 mg g−1 dry weight (dw) in the late ripe stage, while the ratio of reducing sugars to sucrose decreased from 3.45 ± 0.32 to 0.41 ± 0.02. The total phenolic compounds and condensed tannins declined with ripening, particularly during the transition from the breaker to green pedicel stage. Eighteen polyphenols were identified and quantitated, with catechins and hydrolyzable tannins being dominant until the onset of fruit coloration. The transition to the green pedicel stage signaled a precipitous decline (90.9%) in catechins, hydrolyzable tannins (60.2%) and flavonol glycosides (52.1%) concomitant to the rise in gallic acid, which was putatively fueled by the enzymatic hydrolysis of gallotannins in immature fruit. Catechins, hydrolyzable tannins and flavone glycosides were more abundant at higher altitudes and gallic acid at lower altitudes. An antioxidant capacity was also favored by higher elevations and declined with ripening, particularly after the breaker stage. Correlations with FRAP and DPPH assays were significant for the total phenolic content, condensed tannins, catechins and hydrolyzable tannins. The highest correlation factors were obtained for epigallocatechin-gallate (r = 0.920 and r = 0.900; p < 0.01). Although the sharp drop in hydrolyzable and nonhydrolyzable tannins and catechins compromised the in vitro antioxidant capacity at physiological maturity, it also reduced the astringency and configured a palatable organoleptic fruit profile. These changes unraveled significant episodes in the ripening-related secondary metabolism of the carob fruit. They further highlighted the value of immature carob as a potent source of gallotannins, with putative in vivo anti-inflammatory action, and of catechins beneficial in preventing and protecting against diseases caused by oxidative stress.
... Secondary metabolites classes present in carob include phenolic acids, flavonoids and tannins as in Fig. 36.2. Expectedly, polyphenols level depends massively on genetic, environmental factors, extraction process and cultivar type (Loullis and Pinakoulaki 2018;Custódio et al. 2011b). Carob trees of different gender and cultivars revealed that total phenolic content (TPC) were higher in hermaphrodite trees than female ones recording 41.3 mg GAE/g and 18 mg GAE/g, respectively (Custódio et al. 2011b). ...
... Expectedly, polyphenols level depends massively on genetic, environmental factors, extraction process and cultivar type (Loullis and Pinakoulaki 2018;Custódio et al. 2011b). Carob trees of different gender and cultivars revealed that total phenolic content (TPC) were higher in hermaphrodite trees than female ones recording 41.3 mg GAE/g and 18 mg GAE/g, respectively (Custódio et al. 2011b). The content of catechins in such samples ranged from 6-10 mg/g while gallic acid ranged from 1-2 mg/g of different Portuguese cultivars (Custódio et al. 2011b) in contrary to other previous studies recorded gallic acid as the major phenolic acid (Papagiannopoulos et al. 2004;El Ansari Zineb et al. 2017;Corsi et al. 2002;Owen et al. 2003;Ayaz et al. 2007) and suggestive for growing habitat impact on polyphenols composition in carob. ...
... Carob trees of different gender and cultivars revealed that total phenolic content (TPC) were higher in hermaphrodite trees than female ones recording 41.3 mg GAE/g and 18 mg GAE/g, respectively (Custódio et al. 2011b). The content of catechins in such samples ranged from 6-10 mg/g while gallic acid ranged from 1-2 mg/g of different Portuguese cultivars (Custódio et al. 2011b) in contrary to other previous studies recorded gallic acid as the major phenolic acid (Papagiannopoulos et al. 2004;El Ansari Zineb et al. 2017;Corsi et al. 2002;Owen et al. 2003;Ayaz et al. 2007) and suggestive for growing habitat impact on polyphenols composition in carob. Variability among carob phenolic contents of Sicilian origin (Avallone et al. 1997) versus Anatolian one was reported (Ayaz et al. 2007). ...
Ceratonia siliqua (Carob); is a Mediterranean legume globally recognized for its commercial value, being used as a cold beverage, in bakery and confectionary products. It is widely used as a Cocoa substitute not only due to its richness in sugar but rather the absence of caffeine and theobromine stimulant action. Both fruit pulp and seeds are of potential nutritive and medicinal values. The pulp comprises a high sugar content dominantly sucrose as well as polyphenols viz. phenolic acids, flavonoids and tannins. Seeds potential usage is attributed to its locust bean gum (LBG), commercially and pharmaceutically used as gelling and stiffening agent. Carob syrup is a traditional product native to the Mediterranean region, enriched in D-pinitol sugar of anti-diabetic effect. Considering the diversity of carob active constituents’ classes, a myriad of biological effects is recorded to include antioxidant, anticancer, antimicrobial and anti-hyperlipidemic effects. This book chapter presents up to date information on carob usage and chemistry while providing insight on research questions or applications yet to be addressed.
... 12 The phenolic and antioxidant capacity of carob products depend mainly on the carob variety and technological factors associated to carob pulp processing. 13 The carob roasting is a crucial factor and many chemical reactions occur during this process. On one side, sugar caramelisation and Maillard reaction favour the production of furans, esters and pyrroles that recall the aroma of cocoa. ...
... According to the Brazilian legislation, the maximum level of F + HMF allowed in cachaças is 5 mg hL −1 of anhydrous alcohol, 17 and in the particular case of concentrated rectified grape must, EC Regulation No. 1493/99 sets a limit of 25 mg kg −1 of total sugars for HMF. 18 To date, several phenolic and/or antioxidant capacity studies have been done on carob pulp products from different countries: methanolic extracts from Portuguese varietal carobs including Aida, Galhosa and Mulata, 13 mixture of Mulata and Galhosa supercritical carbon dioxide, ultrasound and conventional extracts; 3,19 roasted and unroasted Turkish carob extracts; 7,16 insoluble and soluble roasted Croatian carob fractions after gastrointestinal digestion; 15 Greek carob pod crudes; 20 extracts from German commercial carob products; 9 Lebanese carob-based milk beverage using roasted and unroasted carob from varietal carob pods 2 and methanol:acetone:water (30:30:40 v/v/v) extracts from Polish powdered carob pasta or carob flour. 11 However, information on the factors which are likely to affect the liquor elaboration process is still missing. ...
... (a.2), (b.1), (b.2), (c.1) and (c.2)). Custódio et al. 13 also found differences in TPC for the same two groups of differentiated varietal methanolic extracts, AIDA and Mulata/Galhosa. According to the impact of independent variables and their interactions (Fig. 2(a.1) and (b.1)), particle size and roasting degree are variables with significant influence on phenolic content, while the maceration period was not statistically important. ...
BACKGROUND
The production of the traditional carob liquor from Algarve (Portugal) depends on numerous factors such as carob processing, variety and maceration conditions. An experimental design with 36 runs was created to evaluate the effect of the roasting temperature, particle size, variety of carob and time of maceration on several parameters of carob liquors as gallic acid and total phenolic content, the furanic composition (furfural and 5‐(hydroxymethyl)furfural), browning index and in vitro antioxidant capacity.
RESULTS
The results revealed that carob variety was the independent variable with the greatest effect on antioxidant capacity, total phenolic and gallic acid content. In particular, AIDA liquors presented the highest results, mainly those prepared with unroasted carob. Meanwhile, Galhosa and Mulata liquors showed the greatest concentrations when the carob pulp was roasted at 150 °C. The furanic composition and browning index were greatly influenced by the carob roasting degree.
CONCLUSION
The levels of the main toxic furanics present in carob liquors, furfural and 5‐(hydroxymethyl)furfural, suggest a safe consumption of these beverages even in samples of carobs with the maximum roasting degree. The smallest carob particle size favoured the highest phenolic extraction, while the longest maceration periods decreased the concentration of the toxic furanic compounds studied.
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... Many investigations showed the biological effects of carob mainly attributed to its polyphenolic components known for their antioxidative properties [6,7]. In point of fact, Makris and Kafalas [6] postulated that there was a direct correlation between antioxidant capacity and phenolic compounds of carob pod. ...
... However, in the literature data on carob seeds antioxidant properties related to its polyphenolic composition are limited. In fact Custodio et al. [7] and Durazzo et al. [8] reported correlation between the phenolic content of the carob germ flour and antioxidant activity. To our best knowledge, this is the first study which used aqueous solvents to extract bioactive compounds from whole carob seeds, then quantified phenolic compounds and evaluated the antioxidant activity. ...
... Durazzo et al. [8] by assessing the phenolics content of aqueous organic extract of carob germ, reported high value of total phenolics (71.03 mg/g), another examination of six carob germ cultivars from Portugal showed values (19.8-28.7 mg/g) of total phenolics, (1.9-6.8 mg/g) of tannins and (10.8-18.8 mg/g) of flavonoids [7]. ...
Background
The carob ( Ceratonia siliqua L.) is very old edible fruit, flavorful and often used in foods and beverages. In this present study the composition of phenolic compounds, the antioxidant activity and the production locust bean gum (LBG) of seeds grown in Algeria was explored.
Methods
The phenolic contents (total phenolics, total tannins, total flavonoids, condensed tannins and flavonols) and the antioxidant activity of the aqueous methanolic, ethanolic and acetonic extracts (70, 80 and 80 v/v , respectively) of the seeds were determined using colorimetric methods. Furthermore, the carob bean gum from seeds was extracted and its chemical composition was evaluated.
Results
Aqueous acetonicextract of carob seeds give the highest contents of total phenolics, total flavonoids, flavonols, total tannins and condensed tannins which were 12.24±0.02, 1.33±0.01, 2.97±0.01, 4.29±0.2 and 0.53±0.01 mg/g of dry weight, respectively. The same extract showed also the best antioxidant activity in all assays compared to other extracts.
... A high antioxidative potential and free-radical scavenging activity of phenolic compounds present in pulp, leaves and seeds of carob have also been reported (Kumazawa et al., 2002;Durazzo et al., 2014). Custódio et al. (2011) used pulp extracts obtained from Portuguese cultivars to decrease the viability of different human cancer cell lines. Gallic acid and (+)-catechin acid were the main compounds identified in those extracts, with gender (male, female or hermaphrodite trees) and cultivar significantly influencing the biological activity of the extracts. ...
... The values of total phenols found in ripe fruits agree with those previously reported by Avallone et al. (1997) and Custódio et al. (2011) and significantly lower than those reported by Kumazawa et al. (2002). Such differences may be due to the different methodology used for obtaining phenolic extracts. ...
... In fact, and as reported by Tamir and Alumot (1969), unripe pods are rich in catechins and leucoanthocyanidins, percursors of condensed tannins. The capacity to scavenge free radicals was already reported by Kumazawa et al. (2002) and Custódio et al (2011), although lower activities were reported when compared to our results. In the current work, the capacity of samples to scavenge DPPH free radicals was better than the positive controls (gallic acid and catechin), in contrast to that reported by those authors. ...
The aim of this work was to understand how two different soil types (calcareous - C and non-calcareous soils - NC) affect carob tree nutritional performance and bioactive compounds present in the pulp of the pod. Two orchards were selected in two separately separate locations: in soil C, trees were grown under calcareous conditions, with high percentage of active lime (>9%) and a soil pH of 8.1 and in soil NC, no carbonates were present and soil pH was 6.6. Leaf mineral composition, total leaf chlorophyll and several bioactive compounds were monitored in trees of the cv. ‘Mulata’ during three years and covering two seasons in both orchards. Unripe and ripe pods were separately analyzed. Trees in the NC site were smaller and less vigorous but presented higher leaf chlorophyll, and higher N, Mg, Zn, Fe and Mn in leaves, than trees of the C site. In the latter, only Ca and K were slightly higher. Unripe pods in both orchards showed significantly higher polyphenols content and high anti-oxidant activity (expressed as DPPH). Condensed tannins content was related to leaf mineral composition (N, Zn and Mn) but among these, Mn was the most important and its accumulation in leaves is a good indicator of anti-oxidant activity. Condensed tannins content was higher in unripe pods of trees in the NC site.
... For many years, carob pulp was considered a by-product of the aforementioned industries and its use has been practically restricted to animal feed due to their high content of carbohydrates [sugars (40-60%), mainly sucrose (65-75%)], and dietary fiber and low fat content (0.6%) [2]. However, nowadays there is an increased demand of carob pulp products for human consumption as a substitute or extender for cocoa in different drinks, such as liqueurs and milkshakes, in the preparation of traditional confectionary and baked goods, bread or pasta and as a sweetener for children [2,[4][5][6]. This is mainly due to the previously mentioned composition and its lower content of alkaloids (caffeine and theobromine) and calories than chocolate [1,2,4]. ...
... The carob pulp's dietary fibres and polyphenols have been found to have health beneficial effects [4]. Recent studies have attributed antioxidant properties to the carob pod due to the presence of polyphenols [1,2,4,5]. ...
... The three varieties under study, Galhosa, Mulata and AIDA, were chosen because they are the main Portuguese cultivars present in orchards in Portugal [5]. In terms of the influence of the variety on the concentration of TP and the antioxidant capacity, it can be seen ( Table 2 and 3) that Galhosa liquors presented the lower values (F-C method: between 210.83 ± 9.79 and 766.72 ± 0.66 mg GAE/L; FRAP assay: between 1.57 ± 0.13 and 9.47 ± 0.55 mM AAE and DPPH assay: between 49.52 ± 4.90 and 90.39 ± 0.21% inhibition) closely followed by Mulata liquors (F-C method: between 202.68 ± 5.05 and 942.81 ± 42.87 mg GAE/L; FRAP assay: between 2.56 ± 0.02 and 11.54 ± 0.71 mM AAE and DPPH assay: between 49.24 ± 3.21 and 90.55 ± 0.33% inhibition), whereas AIDA was the variety that showed the highest TP and AC (FRAP method) values (F-C method: between 460.52 ± 1.62 and 1205.69 ± 68.16 mg GAE/L; FRAP assay: between 6.09 ± 0.54 and 23.48 ± 1.64 mM AAE and DPPH assay: between 41.26 ± 0.45 and 86.63 ± 0.12% inhibition). ...
Carob liqueur is a traditional alcoholic beverage manufactured by maceration of plurivarietal carobs with plant-based ethyl alcohols. The main purpose of this research was to investigate the influence of the carob variety and different pod processing processes, on the antioxidant capacity (AC) and total phenolic content (TPC) of the final liquor (fruit macerated in 45% v/v of alcohol). For this, pods from three carob varieties characteristic from the Algarve region were tested: Galhosa, Mulata and AIDA. Fig distillate was the fruit-based ethanol used. The maceration time (one or three weeks), the particle size (flour or pieces) and three roasted degrees (unroasted, 120 °C and 150 °C), were the studied processing variables. Ferric Reducing Ability of Plasma (FRAP) and DPPH free radical scavenging methods were used to measure the AC of the liquors and the TPC was determined by the Folin-Ciocalteu method.
... The seeds from carob pods are mainly utilized for the production of locust bean gum (LBG, E-410), which added as thickening agent and stabilizer in foods. The pulp has been used for animal feed, but recently researchers are focused on its utilization as a valuable food ingredient [29,30,32,33]. ...
... Other hydroxybenzoic acids found are ellagic, syringic, 4-hydrobenzoic, gentinsic and vanillic. Hydroxycinnamic acids are present in relatively low concentrations and include cinnamic, coumaric, ferulic, chlorogenic and caffeic acids [33,[86][87][88][89]. The flavonoid fraction consists of flavonols, flavan-3-ols, flavones, flavanones and isoflavanones. ...
... Hence, substitution of cocoa powder by carob powder is suggested as an effective approach to decrease caffeine and theobromine to trace levels in confectioneries, beverages and baking products [103,104]. Theophylline has been detected in the carob pulp extracts of certain carob cultivars, while it is absent in others [33]. The presence of theophylline should be further investigated in additional carob cultivars as well as in carob products. ...
Cocoa originates from the beans of the cocoa tree (Theobroma cacao L.). It is an important commodity and the main ingredient in chocolate manufacture. Its value and quality are related to complex flavors and to its distinct sensory properties. The increasing demand for cocoa and its rising price urges the research for cocoa substitutes. A potential substitute for cocoa is carob. Carob is the fruit of an evergreen tree (Ceratonia siliqua L.) cultivated in the Mediterranean area, well known for its valuable locust bean gum and also for carob powder and syrup that are obtained from carob pulp. Cocoa beans and carob pods contain various phytochemicals including polyphenols, proteins and amino acids, fatty acids, carbohydrates and fiber. Phytochemicals represent an important source of nutrients and compounds that are beneficial to human health. In this review, phytochemicals in cocoa beans and carob pods and their impact on human health are reviewed. The bioactive compounds that are present in carob, in conjunction with the cocoa-like flavors and unique sensory properties that are enhanced by carob powder roasting, underline carob’s potential to substitute cocoa in various food products. These food applications are discussed in this review.
... One class of compounds that has been the focus of several studies regarding carob, is polyphenols [16][17][18][19][20]. Polyphenols are phytochemicals that contribute to the improvement of human health and prevention of numerous diseases, including cardiovascular diseases, oxidative stress and cancer, and exhibit neuroprotective properties due to their antioxidant and anti-inflammatory activities [21]. Carob polyphenolic extracts have been demonstrated to display anti-proliferative and apoptotic activity against cancer cells, in addition to antidiabetic, anti-diarrheal and anti-hyperlipidemic effects [22][23][24][25][26][27]. The major polyphenols present in carob include free, bound and conjugated forms of phenolic acids, flavonols, flavan-3-oles, hydrolysable tannins and proanthocyanidins with significant biological activities [2,17,23,24,26]. ...
... Carob polyphenolic extracts have been demonstrated to display anti-proliferative and apoptotic activity against cancer cells, in addition to antidiabetic, anti-diarrheal and anti-hyperlipidemic effects [22][23][24][25][26][27]. The major polyphenols present in carob include free, bound and conjugated forms of phenolic acids, flavonols, flavan-3-oles, hydrolysable tannins and proanthocyanidins with significant biological activities [2,17,23,24,26]. The polyphenolic content of carob powder can be influenced through competing pathways upon roasting, as the thermal degradation of phenolic compounds can lead to decreased antioxidant activity, while higher extractability of phenolic compounds from the complex food matrix upon heating has been suggested to increase availability and, consequently, to induce the opposite effect on antioxidant activity [11,12,17]. ...
Recent studies have highlighted the potential of utilizing carob kibbles as a bioactive-rich food ingredient associated with substantial health benefits. Roasting is a key process in enhancing the sensory characteristics of carob kibbles, also affecting the bioactive polyphenols and leading to the formation of Maillard reaction products (MRPs), including the polymeric melanoidins that are associated with a high antioxidant potential but remain unexplored in carob. In this work, we employed for the first time attenuated total reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy to probe the dynamic chemical and structural changes upon the roasting of carob kibbles, along with the investigation of the in vitro antioxidant activity through the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and the determination of the total polyphenolic, proanthocyanidin, gallic acid and cinnamic acid contents. Roasting significantly enhanced the in vitro antioxidant activity of the polyphenolic carob extracts, with different rates at distinct roasting temperatures. The ATR-FTIR analysis enabled the identification of the changes in the structural features of polyphenolic compounds that were related to the improved antioxidant activity upon roasting. Furthermore, the detection of characteristic signatures for the polymeric melanoidins in the infrared (IR) fingerprint region provided the first evidence for the formation and structural properties of these complex, diverse compounds in roasted carob kibbles.
... Fructose Glucose Sucrose Total Sugars Phenolics (g 100 mL −1 ) (g 100 mL −1 ) (g 100 mL −1 ) (g 100 mL −1 ) (mg L −1 GAE) In the conventional heat-assisted method (HAE), variation in phenolics concentration was determined mainly by source material and temperature, while variation in total sugars concentration was not affected by temperature (Table 1). Variation in phenolics content foremost and sugars secondarily was observed in previous studies and attributed to genetic and environmental factors, harvest maturity, and postharvest storage [11,[31][32][33]. Lefkaritiki delivered a juice higher in both sugars and phenolics than Mavroteratsia. ...
... Given that carob constitutes an underutilized tree crop largely limited to cultivation in marginal lands of highly alkaline, calcareous, and infertile soils, where it subsists strictly under rainfed conditions, the impact of agro-environment on the phenolic content of the pod is unsurprising. Further research is therefore warranted to investigate the significance of growth environment on the actual pod content and ultimately juice extract concentrations of sugars and especially phenolics, variation in which can significantly influence the biological activity of carob kibble products [32]. ...
Aqueous extraction of carob kibbles is the fundamental step in the production of carob juice and carob molasses. Improving the theoretical yield in sugars during organic solvent-free aqueous extraction is of prime interest to the food industry. Collateral extraction of phenolics, however, must be monitored as it influences the sensory and functional profile of carob juice. We presently examined the impact of source material, kibble size, temperature, and duration on the efficiency of extracting sugars and phenolics aqueously by conventional heat-assisted (HAE) and ultrasound-assisted (UAE) methods. Source material was the most influential factor determining the concentration of phenolics extracted by either method. Source material also influenced the relative proportions of sucrose, glucose, and fructose, which may impact the perceived sweetness of the juice. Kibble size (medium size M = 9-13 mm; powder size P = 1-4 mm) was more influential with UAE than HAE for both sugars and phenolics but was rendered less influential with prolonged UAE duration. Increasing HAE temperature (from 25 • C to 75 • C) favored the extraction of phenolics over sugars; however, prolonging extraction at 25 • C improved sugar yield without excessive yield in phenolics. Disproportionate extraction of phenolics over sugars limits the use of heat-assisted extraction to improve sugar yield in carob juice production and may shift the product's sensory profile toward astringency. Prolonged extraction at near ambient temperature can, however, improve sugar yield, keeping collateral extraction of phenolics low. Ultrasound agitation constitutes an effective means of extracting sugars from powder-size kibbles. Industrial application of both methodologies depends on the targeted functional and sensory properties of carob juice.
... This pulp is extracted from the carobs, the fruits of carob tree (Ceratonia siliqua L., Fabaceae), one of the most useful Mediterranean trees [1]. Previous works emphasize the beneficial contribution to human health of compounds from carob pulp, including various nutrients (such as several important minerals and vitamins) [2], as well as phenolic compounds with great antioxidant capacity (AC) [3][4][5][6], enzyme inhibitory potential [7], and antiproliferative effect [8], among others. ...
... Phenolics, particularly flavonoids, are compounds linked to the antioxidant capacity (AC) of plant extracts and already found in different carob products [4][5][6]. As can be seen in Table 3, high correlations were found between the total phenolic content (TPC) and the AC by two methods based on different mechanisms used to evaluate the antioxidant effect of phenolic compounds, the ORAC and TEAC assays. ...
Carob liqueur is a traditional Mediterranean alcoholic beverage obtained via a wide range of production techniques contributing to the different organoleptic attributes of the final product. The aim of this research was to evaluate the stability of the chemical composition and biological capacities (antioxidant and enzyme inhibition) under in vitro simulated gastrointestinal digestion of liqueurs prepared by flavouring the fig spirit with carob pulp by maceration, distillation, percolation, or aqueous and hydro-alcoholic infusions. For this purpose, the phenolic and furanic compositions, the total phenolic (TPC) and flavonoid (TFC) contents, antioxidant capacity (AC), and enzyme inhibitory potential against acethylcholinesterase, tyrosinase, α-glucosidase and α-amylase enzymes were evaluated. The content of gallic acid decreased after gastrointestinal digestion, while TPC, TFC, and AC significantly increased after each digestion phase. Overall, no significantly different enzyme inhibitions (p < 0.05) were observed among digested liqueurs, with moderate inhibition against acethylcholinesterase and tyrosinase (enzymes related with neurodegenerative diseases), and potent and low inhibitory capacities for α-glucosidase and α-amylase, respectively (ideal conditions employed in antidiabetic therapy). The study indicates that hydro-alcoholic infusion and maceration were the most appropriate methods to obtain liqueurs with higher values of the aforementioned parameters and safe levels of toxic furanics.
... Several authors investigated and characterized different selections of natural and cultivated carob trees in different Mediterranean countries depends on morphological characters ( [9], [10], [11], [12]) chemical contents ( [13], [14]) and genetic variation ( [1], [9], [10], [15], [16]). ...
... Morphological characters of pods and seeds are the most valuable and quantitative marker widely used to identify Carob varieties ( [4], [17]). Carob pods are characterized by a high sugar content (48-56%) ( [7], [18]), and is a good source of protein, fiber and minerals ( [14], [19]), the different rates of carob chemical composition are depending on the geographical origin, climate conditions and mainly on the genotype ( [3], [11]). ...
... Origin, blending, roasting degree and grinding of coffee beans as well as the brewing influence AO capacity of coffee beverage (Ludwig et al. 2012). In order to imitate coffee and provide caffeine-free beverages without the adverse psychoactive effects of caffeine, certain grains and fruits are roasted and brewed in the same manner as coffee AO capacities of coffee substitutes and their ingredients such as carob (Sahin et al. 2009;Custodio et al. 2011), chicory (Jurgonski et al. 2011), chickpea (Segev et al. 2012), roasted barley (Omwamba and Hu 2010) and fig (Vinson et al. 2005) were reported. ...
... This substitute possesses superior AO activity according to all assays applied. The high AO activity of carob observed previously was explained by the presence of catechin and gallic acid (Custodio et al. 2011). Increasing of AO activity with roasting temperature and duration was reported earlier (Sahin et al. 2009). ...
Antioxidant (AO) capacity of instant, espresso, filter and Turkish/Greek coffee brews, coffee substitutes (roasted chicory root, barley, pea, chickpea, carob and dried fig) and individual compounds (phenolic acids, flavonoids, methylxanthines, N-methyl pyridinium and HMW melanoidins) was assessed using DC polarographic assay based on decrease of anodic current originating from hydroxo-perhydroxo mercury complex formed in alkaline solutions of H2O2 at potential of mercury dissolution, as well as three spectrophotometric assays (DPPH, ABTS and FRAP). A large difference between applied assays ability to recognize various types of individual AOs was noticed. Only according to DC polarographic assay significant AO activity was ascribed to methylxanthines and N-methyl pyridinum. The total content of phenolics (TPC) present in complex samples was determined by FC assay. The highest TPC was ascribed to instant coffees and coffee substitutes while the lowest to decaffeinated filter coffee. Complex samples were grouped based on principal components analysis, phenolics AO coefficient, calculated as the ratio between AO capacity and TPC, and relative AO capacity index (RACI), calculated by assigning equal weight to all applied assays including FC. The highest values of RACI were ascribed to instant coffee brews, followed by substitutes while the lowest to the decaffeinated espresso coffee.
... Recently, it has been reported that carob induces a variety of pharmacological actions including antimicrobial, anti-inflammatory, anti-diarrheal, antiulcer, antioxidant, gastroprotective, and anti-constipation effects . It has ant-diabetic (Qasem et al. 2018), hepatoprotective Martić et al. 2022), and antioxidant and cytotoxic activities effects (Custodio et al. 2011;Ayache et al. 2020). It has also been found to be effective in neurodegenerative diseases (Lakkab et al. 2018) and has a protective effect on colon adenoma cells from the genotoxic impact of H2O2 (Klenow et al. 2009). ...
Doxorubicin (DOX) is an anti-neoplastic therapy, but its use is limited by its deleterious toxic effects including nephrotoxicity and cardiotoxicity. This work aimed at assessing the potential protective effect of Ceratonia siliqua methanol extract (CME) on DOX-induced nephrotoxicity in 5 groups of Wistar rats. Nephrotoxicity was induced experimentally by intraperitoneal (IP) injection of DOX (15 mg/kg). DOX increased serum creatinine, urea, sodium, and potassium levels. It elevated MDA levels in the renal tissue but decreased the concentration of GSH and the activity of GST, CAT, and SOD. Meanwhile, it decreased the level of immunomodulatory anti-inflammatory mediators: IL-10 and TGF-β, as well as the activity of MPO but increased the level of IL-6, TNF-α, and caspase-3 in the renal tissue. DOX has upregulated COX-2, caspase-9, and Bax gene expression and downregulated the Bcl-2 gene expression. Immunolabeling of renal tubular epithelium in DOX-intoxicated rats was moderate to strong against Bax, COX-2, and NF-kβ and weak against Bcl-2. Treatment with CME significantly restored the levels of kidney function parameters and the levels of oxidative stress markers. It stimulated the production of IL-10 and TGF-β and decreased the level of IL-6 and TNF-α. CME reverted the gene expression of COX-2, caspase-9, and Bax. Microscopically, CME alleviated the DOX-induced renal damage. Phytochemical analysis revealed the presence of 26 compounds in the CME. No signs of acute toxicity were recorded by CME up to 4000 mg/kg b. wt. orally into mice. Finally, CME could effectively alleviate the deleterious effects of DOX on the kidney. The safety of carob extract encourages its use in the preparation of valuable therapeutic agents.
... Indeed, the sweet pulp of the carob has been used as cattle feed for a long time (Ait et al., 2007). In addition, fruits and leaves of the carob tree contain proteins and phenolic compounds, and a wide range of physiological activities that include antioxidant, antimutagenic, anticarcinogenic, and antiinflammatory properties (Custódio et al., 2011). Due to their some interesting properties, the pulps, and the seeds have been also used in the pharmacological industry (Makris and Kefalas, 2004). ...
This study was carried out to determine the phytochemical profile and antimicrobial activity of the carob, Ceratonia silique L., (Cs). Mature fruits of Cs trees were randomly collected, and dried in grounded into fine powder. Phytochemical, amino acids, and antimicrobial activity analyses were then applied. The Cs fruits displayed high amounts of terpenoids and moderate amounts of flavonoids and steroids. The pods contained seventeen amino acids, including the nine essential amino acids (histidine, valine, threonine, lysine, isoleucine, leucine, phenylalanine, arginine, and methionine). The remaining eight amino acids were nonessential. Serine displayed a low level (0.63 µg/ml), whereas proline was the most abundant amino acid (232.38 µg/ml). The ethanol extracts of CS pods at a concentration of 100% displayed noticeable activity against the microorganisms utilized in this study (Bacillus subtilis, Staphylococcus aureus, Pseudomonas, Klebsiella pneumonia, Streptococcus, and Escherichia coli). Active inhibition was also observed when using 80% concentration for all organisms except Escherichia coli, Pseudomonas, and Bacillus subtilis. It is concluded that the extracts of Cs can be considered a good source of amino acids. The plant also had significant antibacterial activity. Nabaa et al.
... Ceratonia siliqua (carob), a member of Leguminosae family, is broadly cultivated in Mediterranean countries, and in some parts of the USA and Australia [59]. Previous reports have revealed antimicrobial, antioxidant, and anticancer effects of carob [60]. ...
Purpose
Toxoplasmosis is a globally widespread parasitic disease which causes major health problems in human and animals. This research was conducted to assess the effect of some Egyptian herbal extracts against Toxoplasma gondii ( T. gondii) tachyzoites in vitro.
Methods
The methanol extracts of Withania somnifera, Cyper rotundus , Acacia nilotica , Chrysanthemum cinerariae folium , Anethum graveolens , Raphanus sativus , Ceratonia siliqua , Elettaria cardamomum and Cuminum cyminum were tested against T. gondii tachyzoites.
Results
Among the tested plants, the extracts from Raphanus sativus, Cuminum cyminum, and Ceratonia siliqua exhibited high anti-Toxoplasma activities at 50 µg/ml, relative to sulfadiazine. They showed low IC 50 values on T. gondii (7.92, 9.47 and 13.52 µg/ml, respectively) and high selectivity index values (100.79, 59.19, and 29.05, respectively). Scanning electron microscopy (SEM) findings indicated evident morphological changes in tachyzoites treated with these three herbal extracts.
Conclusion
Raphanus sativus , Ceratonia siliqua , and Cuminum cyminum methanol extracts could be promising sources of new medicament for toxoplasmosis.
... The total phenolic content (TPC) of carob pulps was 18 to 41.3 mg gallic acid equivalent (GAE)/g. Catechins and gallic acid contents ranged from 6-10 and 1-2 mg/g respectively [10]. The pods comprised 3.7 mg/g total tannins [4]. ...
... mg/ 100 g dw. In another study examining six carob seed varieties from Portugal, the total amount of phenolic substance was found to be between 19.8-28.7 mg/g (Custodio et al., 2011). Brahim et al. (2017) investigated total phenolic content of carob pods (pulp and seeds) and leaves of different ages. ...
... Therefore, the detection of gallic acid and gallotannins in the aqueous extracts of carob pods collected from Italy was contributed to a marked reduction of the carcinoma T1 cell proliferation [10]. Likewise, it has also be demonstrated a high inhibitory capacity of methanolic extract obtained from Portuguese carob pods on HeLa cells viability [11]. This was attributed to the presence of catechin and gallic acid on such extract. ...
Ceratonia siliqua L. pods have been classically used in Mediterranean folk medicine as a potential source of natural antibacterial and antioxidant agents. The in vitro antiradical, immunostimulant, antibacterial and cytotoxic properties, as well as phytochemical composition of the ethanolic and aqueous extracts obtained from carob pods harvested at three ripening stages were assessed. Fructose, glucose, and sucrose, as quantified in both extracts, showed varied yields according to the ripening stage and solvent type. Likewise, significant differences were shown in bioactive compounds content depending on extraction solvent and ripening stage. Moreover, gilthead seabream (Sparus aurata L.) leucocyte viability and functions were not altered after being incubated for 24 h with the extracts of carob pods. On the other hand, the ethanolic extracts (0.75 and 1 mg mL⁻¹), exhibited an interesting cytotoxic effect on a tumoricidal fish cell line (PLHC-1 cell). Besides, the ethanolic extract (1 mg mL⁻¹) displayed a moderate bactericidal activity against different examined bacteria. Photobacterium damselae was found to be the most sensitive to different carob pod extracts, except for the aqueous extract obtained from ripened pods. While Vibrio harveyi and Vibrio anguillarum exhibited a strong resistance against the aqueous extracts. Our data indicate that the carob pods are an excellent source of natural bioactive molecules that would be used to prevent and treat several marine animal diseases.
Graphical Abstract
... The difference of these results can be explained by the use of different extracts, containing completely different bioactive compounds, besides the use of a different assay for detecting living cells by MTT oxidation and not considering apoptotic cells. In accordance with our findings, carob pod extracts reveal a cytostatic activity by arresting breast cancer MDA-MB-231 cells in the S phase, with 30% of the cells in sub-G1 (apoptosis) at 40 mg/mL [51]. Nevertheless, the literature demonstrates that carob pods exhibit a strong antiproliferative capacity on several cancer cell lines, different from those investigated here. ...
The present work describes the volatile compounds profile and phytochemical content of Ceratonia siliqua L. Fifty different components have been identified. Among them, three constituents are shared i.e., 2-methlybutanoic acid, methyl hexanoate and limonene by different common carob preparations: pulp decoction (PD), seeds decoction (SD) and Rob, a sweet syrup extracted from the pulp of the carob pod. Each extract exhibits different volatile aromatic emission profiles. The antioxidant activity of the extracts was evaluated using three methods, DPPH, ABTS and FRAP, producing a dose-dependent response. The IC50, when determined by FRAP, gave the lowest values (0.66 ± 0.01, 0.73 ± 0.05 and 0.55 ± 0.00 mg/mL PD, SD and Rob, respectively). The nociception essay, after intraperitoneal injection of acetic acid in mice, demonstrated that Rob, pulp and seeds decoction extracts showed an efficient inhibition of writhes over time, with persistence over 30 min. The SD decoction revealed the highest efficacy in decreasing the writhing reflex (90.3 ± 1.2%; p < 0.001). Furthermore, the proapoptotic activity of SD against three human cell line, THP-1, MCF-7 and LOVO, evaluated by flow cytometry, showed a significantly stronger proapoptotic activity on colon cancer (LOVO) than on the other cell lines, a phenomenon known as phenotypic selectivity.
... [6][7][8][9][10] Consumption of carob pulp was also related to several health promoting effects: it exhibits high antioxidant activity, improves glucose tolerance and attenuates cholesterol levels. [11][12][13][14][15][16] The chemical composition of the pulp strongly depends on the cultivar, origin and harvesting time. 3,17 Due to the high free sugar content (>48% of which more than 65% sucrose), the pulp is gaining popularity as a natural sweetener in foods, with a flavor profile and appearance similar to cocoa powder. ...
Carob is the fruit obtained from Ceratonia siliqua L. and it is a source of bioactive compounds that have been linked to several health promoting effects, including lowering blood cholesterol concentration. The objective of this study was to connect the physicochemical changes of carob flour occurring during roasting with its capacity to bind glycoconjugates of bile acids. Carob flour samples were roasted for different times at 150°C and chemically characterized by measuring the concentrations of tannins and polyphenols. Data showed that carob flour binds high amounts of bile acids: 732.6 µmol of bound bile acid per g of carob flour which is comparable to the 836.2 µmol per g bound by cholestyramine, a known cholesterol lowering drug. The carob flour ability to bind cholesterol decreases up to 40% during roasting. Data suggested that tannins and insoluble components play a major role in binding bile salts, as a result of hydrophobic interactions.
... In the present study, autoclaving treatment increased the DRSC of Mugua compared to the single reflux method (Table 3), which was opposite to the change of total phenolics from Mugua, and opposite to other studies as the total phenolics always have a significant correlation with DRSC. [23] However, total phenolics of Mugua have been shown with no significant correlation with DRSC, which may contribute to the phenolic composition in Mugua, such as the anthocyanins have no significant correlation with DRSC. [24,25] On the other hand, the content of polysaccharides was increased by hyphenated method, which has antioxidant activity, and monosaccharides are reductive agents as they can supply or donate hydrogen, which can combine with radical and form a stable radical to terminate the free radical chain reaction. ...
Chaenomeles (called Mugua in China)
is an important material in the food industry, and XuanMugua is the only one which is allowed to serve as a homology of medicine and food in China. This study was conducted to develop an effective extraction method for XuanMugua by autoclaving treatment hyphenated with reflux. Hyphenated extraction method gave 19% higher extraction efficiency, 5% higher content of polysaccharides, and 29% higher content of triterpenes than single reflux. Furthermore, it improved the antioxidant activities of Mugua by increasing DPPH radical scavenging by 0.80 times and similar ABTS and hydroxyl radical scavenging abilities.
... Furthermore, the obtained results agree with previous published data which revealed that carob leaf extracts exhibit antitumor activity against different tumor cell lines. Indeed such leaf extracts (methanolacetone-water) were seen to exert an antitumor effect on human colon cancer cells [18], while a methanolic carob leaf extract decreased the viability of human cervical adenocarcinoma cell line (Hela) in an impressive manner but with less impact on prostate (DU-145), breast (MDA-MB-231) and colon (HCT-166) cell lines [49]. For its part, an aqueous carob leaf extract also induced some antiproliferative effects on a mouse hepatocellular carcinoma cell line [41]. ...
Carob leaves, the main residues of the carob tree, were investigated as a renewable and abundant source of bioactive compounds for fish aquaculture. Aqueous and ethanolic extracts obtained from carob leaves were characterized in terms of biochemical composition, antiradical and cytotoxic effects and immunostimulant and antibacterial activities. The ethanolic extract showed higher levels of total phenolics, flavonoids and condensed tannins and higher antioxidant activity than the aqueous extract. No significant immunostimulant effects were observed on gilthead seabream (Sparus aurata) head kidney leucocytes (viability, phagocytosis and respiratory burst activities and peroxidase content) after incubation for 24 h with different extracts. Furthermore, the ethanolic extracts used at 0.5, 0.75 and 1 mg mL-1 and aqueous extracts at 1 gmL-1 had a cytotoxic effect on PLHC-1 cells. When the bactericidal activity was tested against three fish pathogenic bacteria (Vibrio harveyi, Vibrio anguillarum and Photobacterium damselae) notable activity of the different extracts was detected against P. damselae at all three concentrations. A similar effect was demonstrated against V. haryeri when ethanolic extracts were used in the same range of concentrations. This work demonstrates interesting in vitro effects of carob leaf extracts and suggests it could be used as an alternative to chemical compounds with farmed fish. The concentration and nature of the extracts were very important in terms of any positive results.
... Indeed, the genetic and environmental expressions were known by its considerable effect on enzymatic activities that are involved in the biosynthesis and biodegradation of several primary and secondary metabolites (15). Therefore, pulps collected from Portugal (16) contain higher concentrations of total polyphenols than those collected from Algeria (12) and Tunisia (17). Besides, it has been also proved that the ripening process alters significantly this composition. ...
The Ceratonia siliqua fruits contain several substances known to have high adaptability to environmental conditions. The aim of this paper is to evaluate the changes in physicochemical properties of different Tunisian provenances of carob pulps harvested at three ripening stages. Furthermore, six provenances were investigated during the ripening process in terms of their moisture, ash, minerals, bioactive compounds, antiradical activity and sugar profile. The results demonstrated that all examined parameters are highly influenced by geographic origin. Concerning ripening impact, our data showed that water and ash content significantly decreased during the development of six provenances, as well as the bioactive and mineral contents. Thus, the total polyphenols (TP), total flavonoids (TF) and condensed tannins (CT) contents exhibited the highest levels in the unripe fruits. The antiradical activity trend was positively correlated to the behavior of the bioactive compounds content. Moreover, the sucrose, glucose and fructose were the main sugar qualified and quantified in carob pods at different ripening stages. At the maturity stage, the monosaccharide contents (glucose and fructose) were slightly reduced, while, the sucrose was rapidly accumulated. In conclusion, the ripening process diversely affected the nutritional composition and generally extended the exploitation of carob fruits. The study could provide valuable information about the suitability of carob pods at different maturity stages as potential biomaterials for nutraceutical applications.
... Ancak bitkilerin bileşimlerinde bulunan maddelerin bitkinin çeşidine, yetişme yerine, mevsime göre değişiklik gösterebileceği bilinmektedir (15,16). Custodio ve ark., dişi, erkek ve hermafrodit ağaçlardan hazırladıkları ekstraktların total fenolik içerik miktarı ile gallik asit, mirisetin, metil gallat gibi bazı fenolik bileşenlerin miktarlarını karşılaştırmışlar; hermafrodit ağaçların en yüksek total fenolik içeriğe sahip olduğunu ve fenolik bileşen oranlarının da çeşitler arasında farklılık gösterdiğini bulmuşlardır (17,18). Çalışmamızda, araştırılan fenolik bileşenlerden vanilik asitin hem yaprak hem de meyvelerde en yüksek oranda bulunduğu belirlenmiştir. ...
Background: Ceratonia siliqua L., a member of the family Leguminosae, has been reported to have antimicrobial, antifungal and cytotoxic activity (carob, kharnub). In this study, the antiproliferative and cytotoxic effects of extracts prepared with methanol and ethanol from the carob leaves and fruits on HeLa cells were investigated. Additionally, 3T3, embryonic mouse fibroblast cell line, was used as a control.
Material and Methods: For these purposes, after treatment of the extracts at different concentrations (200-800μg/mL) for 48 hours, viable cell counts (proliferation, MTT method), mitotic index (mitotic activity) and labeling index (DNA synthesis) were determined. In addition, the phenolic component compositions in the extracts were also determined by LC-MS / MS.
Results: According to the findings, the methanolic and ethanolic leaf extracts of carob bean suppressed mitotic activity and proliferation by inhibiting the DNA synthesis of human cervical cancer HeLa cells. However, these extracts also exhibited a cytotoxic effect against normal fibroblast 3T3 cells. Ethanolic fruit extract reduced the DNA synthesis of HeLa cells.
Conclusions: The active substances or preparations obtaining from the C. siliqua plant have the possibility of being an anticancer agent, whereas their cytotoxic effects on the normal cells should not be overlooked.
... A previous study correlated the antioxidant and cytotoxic activities of carob to its high phenolics content. Nevertheless, carob pods were more enriched with catechin than gallic acid contrary to our results (Custódio et al., 2011) which reflects the influence of the geographical origin. Interestingly, the relative content of gallic acid levels varied among geographical sources even inside Egypt localities, with unripe fruits recording the highest percentile in Giza specimens (7%) versus that of Behira and Siwa ca.5% in both samples. ...
Ceratonia siliqua is a legume tree of a considerable commercial importance for the flavor and sweet industry. In this context, it is cultivated mostly for its pods, which are known for their nutritive value and multiple health benefits. However, metabolite patterns, underlying these properties are still mostly uncharacterized. In this study, the role of geographical origin, ontogenetic changes and thermal processing on Ceratonia siliqua pod
metabolome was assessed by mass spectrometry (MS)-based metabolomics. Thereby, a total of 70 fruit primary metabolites, represented mainly by carbohydrates, organic and amino acids were detected. Analysis of secondary bioactive metabolites assessed by ultra-high-performance liquid chromatography-electrospray ionization high resolution mass spectrometry (UHPLC-ESI-HR-MS) revealed in total 83 signals. The major signals, most significantly contributing in discrimination of C. siliqua specimens were assigned to tannins and flavonoids. PCA models derived from either UHPLC-MS or GC-MS proved to be powerful tools for discrimination of C. siliqua specimens
... The endosperm of carob seeds produces a gum (carob bean gum) rich in galactomannans, used as a food additive [3]. In addition, the seed coat and pod pulp are used in the food industry to produce locust bean gum (LBG) and natural antioxidants [11,12,40]. Carob pods are also used in the pharmaceutical and cosmetic industries [2,32,40] and could provide an economical source of bioethanol in the future, particularly in arid regions [40,51,65]. ...
Ceratonia siliqua L. is a slow growing evergreen tree of the family Fabaceae used for the rehabilitation of marginal and submarginal dry areas of the Mediterranean basin due to it's resistant to drought and salt tolerance. In this study, the effects of different basal media (Woody Plant Medium and Murashige and Skoog medium), explant types (cotyledon and hypocotyl) and growth regulators (BA, Kinetin and NAA) on in vitro callus formation, differentiation of callus to shoot and root formation were investigated. High frequencies of caullogenesis were obtained and the best medium for callus induction was WPM supplemented with 1.0 mg L-1 BA + 0.5 mg L⁻¹ Kinetin + 0.5 mg L⁻¹ NAA and 0.5 mg L⁻¹ BA + 1.0 mg L⁻¹ Kinetin + 0.5 mg L⁻¹ NAA for hypocotyl explants and 0.5 mg L⁻¹ BA + 1.0 mg L⁻¹ Kinetin + 0.5 mg L⁻¹ NAA; 0.5 mg L⁻¹ BA + 0.5 mg L⁻¹ Kinetin + 0.5 mg L⁻¹ NAA and 1 mg L⁻¹ Kinetin + 0.5 mg L⁻¹ NAA for cotyledon explants. Callus induction was more readily obtained from hypocotyl explants than cotyledon explants. It was determined that explant types as significant on shoot formation, statistically. The shoot ratio was obtained from cotyledon explants in WPM as 10%. The best regeneration was obtained from cotyledon explants placed on WPM (30%) instead of MS medium.
... Carob is the beanlike fruit of Ceratonia siliqua, which grows in many parts of the Mediterranean region [29] and in some regions of USA, Latin America and Australia [30]. Due to a high level of tannins that can cause astringency, the use of the whole fruit in human consumption is rather limited [31]. ...
In the present study, we have investigated the effects of polyphenol-rich infusions from carob leaves and OFI-cladodes on inflammation associated with obesity and dextran sulfate sodium (DSS)-induced ulcerative colitis in Swiss mice. In vitro studies revealed that aqueous extracts of carob leaves and OFI-cladodes exhibited anti-inflammatory properties marked by the inhibition of IL-6, TNF-α and nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells concomitant with NF-κβ nucleus translocation inhibition. For in vivo investigations, Swiss male mice were subjected to control or high fat diet (HFD). At the 8th week after the start of study, animals received or not 1% infusion of either carob leaves or OFI-cladode for 4 weeks and were subjected to 2% DSS administration in drinking water over last 7 days. After sacrifice, pro-inflammatory cytokines levels in plasma and their mRNA expression in different organs were determined. Results showed that carob leaf and OFI-cladode infusions reduced inflammation severity associated with HFD-induced obesity and DSS-induced acute colitis indicated by decrease in pro-inflammatory cytokines expression (as such TNF-α, IL1b and IL-6) in colon, adipose tissue and spleen. In addition, plasma levels of IL-6 and TNF-α were also curtailed in response to infusions treatment. Thus, carob leaf and OFI-cladode infusions prevented intestinal permeability through the restoration of tight junction proteins (Zo1, occludins) and immune homeostasis. Hence, the anti-inflammatory effect of carob leaves and OFI-cladodes could be attributed to their polyphenols which might alleviate inflammation severity associated with obesity and colitis.
... They are more important in food industry and are a source of many products such as gum, sugar, and alcohol [11] and as a natural additive (E 410) [12]. The different parts of carob tree exerted diverse physiological functions [13][14][15][16][17][18][19][20]. ...
Carob tree, Ceratonia siliqua L., is a medicinal plant used in Tunisian traditional medicine for the treatment of the gastro-intestinal (GI) disorders. In this respect, a relatively large number of scientific publications on the carob tree have been published in recent years. Therefore, the present review was aimed to analyze the traditional uses, phytochemical constituents and pharmacological activities of Ceratonia siliqua on the GI tract. Indeed, previous investigations on the carob pods and leaves have revealed the presence of a number of compounds including high amounts of carbohydrates, dietary fibers, minerals, polyphenols, flavonoids and low amounts of protein and lipids. This plant possesses anti-inflammatory, antimicrobial, anti-diarrheique, antioxidant, anti-ulcer, anti-constipation and anti-absorptive of glucose activities in the gastrointestinal tract. Based on the chemical and pharmacological characteristics of C. siliqua, we concluded that this species has beneficial preventive and therapeutic properties, especially, in digestive tract.
... 25 Leaves and pods of carob exerted diverse physiological functions such as its antioxidant activity. [26][27][28][29][30][31][32][33] Indeed, the extracts of the carob tree showed signicant radical scavenging activity 34 and a remarkable ability to inhibit tumor cell proliferation. 26 Carob tree extracts contain anti-proliferative agents that could be of practical importance in the development of functional foods and/or chemopreventive drugs. ...
... 25 Leaves and pods of carob exerted diverse physiological functions such as its antioxidant activity. [26][27][28][29][30][31][32][33] Indeed, the extracts of the carob tree showed signicant radical scavenging activity 34 and a remarkable ability to inhibit tumor cell proliferation. 26 Carob tree extracts contain anti-proliferative agents that could be of practical importance in the development of functional foods and/or chemopreventive drugs. ...
The current study aimed to estimate the chemical composition of the powder of both leaves and seeds of Ceratonia siliqua, which was purchased from the local market in Iraq/Baghdad. Where the test included (moisture-fat-protein-carbohydrate-ash-fiber), both leaves and seeds were extracted by Soxhlet continuous extraction machine using ethanol solvent. Several chemical tests were conducted on the extracts, which included the following: Detection of phenolic compounds and flavonoids by HPLC technology (high-performance liquid chromatography), where the results showed the appearance of several peaks at a wavelength of 280 nm for several phenolic compounds and at different time lengths. The inhibition activity was evaluated for two types of bacteria (positive/gram-negative), one type of yeast, two types of molds, and four concentrations of extracts (25-50-75-100 mg/ml). The results showed that the alcoholic extract of the leaves of the carob plant was superior to the seeds against all microorganisms. As for the antioxidant activity, it was approved by the DPPH test compared to vitamin C, as it is a natural antioxidant. The results showed that both leaf and seed extracts were superior to vitamin C, as the IC50% of seed extract was 7 ppm 11 and leaf extract was 184. As for vitamin C, the percentage of free radical inhibition was 50%, 373PPM, as it is known that the higher the IC50 value, the lower the antioxidant activity.
EU funds are all financial and operative programs funded by the European Union, either in the Member States (Union Programs and instruments of the Cohesion, Agriculture and Fisheries Policies), or outside of them (preaccession funds, other country programmes). In order to develop the regions, cities and villages of Croatia, the programme of strategic investment is established by the regional policy which promotes economic and social growth and improves the living standard. It gives support for less developed areas and rural regions as an expression of solidarity. Purpose of this research paper is to identify level of knowledge about EU funds in rural area with survey method. There is a practical example of a survey of the respondents on the impact of EU funds on reducing rural regional inequalities as a result of this paper.
area with survey method. There is a practical example of a survey of the respondents on the impact of EU funds on
reducing rural regional inequalities as a result of this paper.
The carob tree is a common tree throughout the world. It has received great attention due to its potent biological effects and diverse industrial applications resulting from its richness in phytochemicals, particularly polyphenols. The polyphenol content, the phenolic composition, and the antioxidant activity of carob were studied with variable results. Several factors are involved in this variability, such as gender, variety, cultivar, ripening stage, environmental conditions, and extraction techniques. Also, the bioaccessibility of carob polyphenols is strongly affected by several factors, such as the phenolic fraction, food matrix, structure of compounds, and particle size. As a result, the purpose of this paper is to discuss the various factors of variation as well as the extraction and encapsulation techniques used to improve their recovery and bioaccessibility.
Doxorubicin (DOX) is an effective antitumor therapy but its use is limited by its deleterious toxic effects including nephrotoxicity and cardiotoxicity. The aim of this work was to assess the potential protective effect of Ceratonia siliqua methanol extract (CME) on DOX-induced nephrotoxicity in 5 groups of rats. Rats in groups 1and 2 were given normal saline while groups 3–5 were given Vitamin C (reference antioxidant, 250mg/kg), CME (500mg/kg) and CME (1000 mg/kg) for 5 days. On the 5th day, 1 hour after the last treatment dose, rats of groups 2–5 were given DOX in a dose of 15 mg/kg IP. DOX increased serum creatinine, urea, sodium and potassium and decreased GSH concentration, GST, CAT, SOD and MPO activities but increased MDA. It increased the inflammatory mediators (COX-2, IL-6, TNF-α, and NF-κβ) but decreased the anti-inflammatory cytokine (IL-10) and the Transforming growth factor-β (TGF-β). DOX has up-regulated COX-2, Caspase-3, Caspase-9, Bax and NF- κβ transcripts and down-regulated the anti-apoptotic Bcl-2 as assessed by immunohistochemistry and gene expression analysis. CME significantly improved the levels of kidney function parameters and restored the levels of the oxidative stress markers. It also decreased the level of COX-2, IL-6, TNF-α, and NF-κβ and stimulated the production of IL-10 and TGF-β. CME down-regulated the expression levels of the Bax, Cox-2 and caspases and up-regulated the anti-apoptotic Bcl-2. Microscopically, CME alleviated the DOX-induced renal damage in dose dependent manner. Phytochemical analysis revealed the presence of 26 compounds among which 4 major compounds (over 5%) in the CME. Acute toxicity test revealed that CME is not toxic up to 5 g/kg orally into rats. In conclusion, CME could effectively alleviate the deleterious effects of DOX on the kidney. The safety of carob extract encourages its use in the preparation of valuable therapeutic agents.
Cyclophosphamide is one of the chemotherapeutics for cancer, however this drug has severe side effects in multiple organs and is a well-known mutagenic agent. In this work the protective activity of antioxidant-rich Carob (Ceratonia siliqua L.) pods and leaves extracts against cyclophosphamide induced genotoxicity was studied. Safety of these extracts was evaluated in acute and subacute toxicity tests at a dose of 1500 mg/kg/day in albino rats. Both extracts exhibited no mortality or significant changes in rats' general behavior. Moreover, organ weights and blood chemistry revealed no toxic effects of these extracts (p>0.05).The anti-mutagenic potential of carob pods and leaves extracts were evaluated using in vivo bone marrow chromosomal aberration and micronucleus assays and in vitro comet assay for peripheral blood white cells. Results showed that rats that were treated with cyclophosphamide along with carob pods had a significant decrease in chromosomal aberrations when compared with cyclophosphamide-treated rats (p<0.05). Pods extract also showed significant reduction in cyclophosphamide-induced DNA migration (p<0.05). There were also highly statistically significant differences in the means of micronuclei (MNs) in immature red blood cells between cyclophosphamide group and carob leaves group (p<0.01).It could be concluded that Carob extracts had protective effect against cyclophosphamide- induced genotoxicity and may be considered as anti-mutagenic agent after conducting more studies.
In this work, a simple, reliable, and fast capillary electrophoretic method was developed and validated for the simultaneous determination of 12 polyphenolic compounds, the most frequently found in carob's pulp and seeds. The present work deals with the development of a novel dual electrophoretic system based on the combined use of β‐CD and ionic liquid (IL) as buffer additives. A baseline separation of the target analytes was achieved in less than 10 min by using a BGE consisting of 35 mM borate along with 15 mM β‐CD and 3 mM L‐alanine tert butyl ester lactate (L‐AlaC4Lac) IL as buffer additives at pH 9.5, a temperature of 25°C and an applied voltage of 30 kV. The application of the developed electrophoretic method to real samples enabled the identification and quantification of the main phenolic constituents of both ripe and unripe carob pulp extracts. The results revealed the predominance of gallic acid in both ripe (183.92 μg/g carob pulp) and unripe (205.10 μg/g carob pulp) carob pulp and highlighted the great influence of the ripening stage on carobs polyphenolic composition, with unripe pods being more enriched in polyphenols (total phenolics detected: 912.58 and 283.13 μg/g unripe and ripe carob pulp). This article is protected by copyright. All rights reserved
Polyphenols in carobs have recently attracted great attention due to their wide range of biological and health promoting effects. A comprehensive study was conducted to find an optimum method for the extraction, purification and characterization of these valuable bioactive substances. Under this framework, the ultrasound-assisted extraction (UAE) of polyphenols from carob pulp was optimized by the maximization of the yield in total phenolics using response surface methodology. In particular, the effects of solid-solvent ratio, solvent concentration, extraction time, sonication amplitude, and sonication mode were investigated and optimized using a complete experimental design. In comparison to conventional extraction techniques, UAE offered a higher yield of antioxidants and a shorter processing time. Solid-phase extraction was evaluated as a clean-up strategy prior to the electrophoretic analysis of extracts. The results from the analysis of real samples revealed the predominance of gallic acid and highlighted the great influence of the ripening stage on carobs composition.
Carob (Ceratonia siliqua L.) tree is considered among the most important forest-fruit species native to the Mediterranean region. It has various uses and great valorization potential, all parts of this plant could be exploited as a source of income, as human food or livestock fodder as well as source of raw materials for pharmaceutical, cosmetic, or food industries. Moreover, due to its particular agroecological features, carob tree offers the advantage of growing in poor and unfertile soils in the Mediterranean and Mediterranean-like regions of the world. Thus, carob trees are suitable for the rehabilitation of marginal and sub-marginal areas, helping to compensate for the expanding land desertification in these regions where it can play the role of pioneer and productive species. Carob has been intermittently explored over the last 20 years as a potential tree crop industry in low rainfall areas. The importance of developing the industrial agroforestry potential of the carob tree is hurdled by the lack of options for agroforestry, especially in Mediterranean regions with low rainfall (below 500 mm), and by the need to develop suitable practices for the sustainable management of natural resources. Viable commercial carob cultivation will require mastering efficient farming practices with detailed attention to water requirements and soil fertility. It would improve agricultural productivity in low rainfall areas, help manage water and land degradation, diversify farmers’ incomes, and contribute to the development of export industries contributing to balance the economy of the country. This chapter will provide current knowledge regarding the use of mycorrhizal symbiosis for the improvement of carob culture and productivity in the context of Mediterranean ecosystems. An overview on the multipurpose potential of the carob tree and how spreading its cultivation will benefit people and the environment in marginal areas is highlighted.
We evaluated the cytotoxic and genotoxic effects of protein isolates of ayocote beans on adult male CD1⁺ mice and the anticancer activity of their protein fractions using SiHa Cells. The mice that received protein isolate showed a Polychromatic Erythrocytes (PCE) increase at 48 h; a significant PCE decrease was observed in mice administered the protein isolate-Daunorubicin (isolate-DAU) mix. A micronucleated erythrocytes (MNE) increase was observed in mice that received the mix in all times tested. All protein fractions of ayocote beans (black and purple) showed inhibition against SiHa cells proliferation at doses of 3000 or 5000 µg/mL. The maximal inhibitory concentration (IC50) of each protein fraction was reached at a concentration of 5000 µg/mL. A synergistic effect of isolate-DAU was observed in the in vivo test; the effect of the protein fractions on SiHa cells proliferation depended of the ayocote bean variety used.
Dans la présente étude, nous avons étudié les effets des infusions de feuilles de caroube et de cladodes OFI riches en polyphénols sur l'inflammation associée à l'obésité et la colite ulcéreuse induite par le dextran de sulfate de sodium (DSS) chez les souris suisse.Des études in vitro ont révélé que les extraits aqueux de feuilles de caroube et de cladodes OFI présentaient des propriétés anti-inflammatoires marquées par l'inhibition de la production d'IL-6, de TNF-α et d'oxyde nitrique (NO) dans les cellules RAW 264.7 stimulées par des lipopolysaccharides (LPS). Inhibition de la translocation du noyau NF-κβ.Pour des investigations in vivo, des souris mâles suisses ont été soumises à un régime contrôle (ND) ou à un régime riche en graisses (HFD). A la 4ème semaine après le début de l'étude, les animaux ont reçu ou non 1% d'infusion de feuilles de caroube ou d'OFI-cladode pendant 6 semaines et ont été soumis à une administration de DSS à 2% dans l'eau potable au cours des 7 derniers jours. Après sacrifice, les niveaux de cytokines pro-inflammatoires dans le plasma et l'expression de leur ARNm dans différents organes ont été déterminés. Les résultats ont montré que les infusions de feuille de caroube et de OFI-cladode réduisaient la sévérité de l'inflammation associée à l'obésité induite par HFD et la colite aiguë induite par le DSS indiquée par une diminution de l'expression des cytokines pro-inflammatoires (comme TNF-α, IL1b et IL-6) tissu adipeux et rate. En outre, les taux plasmatiques d'IL-6 et de TNF-α ont également été réduits en réponse au traitement par les infusions. Ainsi, les infusions de feuilles de caroube et d'OFI-cladode ont empêché la perméabilité intestinale grâce à la restauration des protéines de jonctions serrées (Zo1, occludines) et à l'homéostasie immunitaire. Ainsi, l'effet anti-inflammatoire des feuilles de caroube et des cladodes OFI pourrait être attribué à leurs polyphénols qui pourraient atténuer la gravité de l'inflammation associée à l'obésité et à la colite.
Ceratonia siliqua and some species of Prosopis (Fabaceae family) are commonly known as carob trees. The flours obtained from their pods are used in the food industry, as cocoa substitute in the confectionery and also used in beverages and mixed with products derived from cereals. The aim of this study was to compare and characterize the physical and chemical properties, specially the antioxidant activities, of the two commercial carob flours. Commercial Prosopis spp. (mainly from P. alba) flour exhibited high content of protein, starch and fat, while commercial flour from C. siliqua had a lower content of these compounds, but higher antioxidant activity. By nuclear magnetic resonance (NMR) the aqueous extracts of the two carob flours were analysed and concluded that they had similar content of sucrose, but C. siliqua had more monosaccharides and pinitol. This important cyclitol has beneficial physiological effects, improving the glycaemic level and, thus, having a great potential in the food industry. We conclude that the commercial flour of C. siliqua has a better nutritional potential than that of Prosopis spp., owing to dietary fiber, total phenols, pinitol and antioxidant activity. Our results corroborate the nutritional benefits of the commercial supplements already available for healthy food formulations.
The consecutive extraction via different solvents of increasing/decreasing polarity, which allowed a pre-fractionation of the extracted compounds, is widely used as major method for extraction of polyphenols. Thus, the phenolic compounds of Ceratonia siliqua L., at unripe and ripe stages, were extracted by four solvents of increasing polarity (hexane, chloroform, ethyl acetate, and acetone/water (70:30, v/v), consecutively. The extracts were evaluated for their total phenolic and flavonoid contents, while the antioxidant capacity was assessed in vitro using DPPH radical and hydrogen peroxide (H2O2) scavenging assays. The phenolic profile has been carried out by HPLC-MS/MS. The results showed that the phenolic contents and antioxidant capacity varied with the nature of extracts and the ripening stage. The 70% aqueous acetone and the ethyl acetate extracts contained the highest phenolic and flavonoid contents, and subsequently showed a pronounced scavenging activity on DPPH and hydrogen peroxide at unripe and ripe stages. The HPLC-MS/MS allowed the identification of five free phenolic acids (gallic, syringic, cinnamic, p-coumaric and ellagic acid for the first time) and five flavonoids (apigenin, naringenin, kaempferol, quercetin rhamnoside and myricetin rhamnoside) at both ripening stages.
Le caroubier est considéré comme l'un des arbres fruitiers et forestiers qui présente le plus grand potentiel de valorisation puisque toutes les parties de cette plante sont utilisables dans plusieurs applications industriels. Le caroubier est cultivé dans plusieurs régions du Liban mais peu d'études sont disponibles sur les voies de valorisation et sur les propriétés fonctionnelles et structurales des graines et des gousses. Ainsi, dans le cadre d'une démarche qui vise à promouvoir et contribuer à une meilleure valorisation et gestion de cette ressource renouvelable, nous avons entrepris dans cette thèse des travaux consacrés à la caractérisation et à l'évaluation de la valeur nutritive et fonctionnelle des gousses de caroube issues de plusieurs régions libanaise. Ces travaux ont porté notamment sur la caractérisation morphologique et physico-chimique des gousses et sur la purification des gommes extraits des graines. Ces gommes ont fait l'objet d'une étude approfondie portant sur leur comportement rhéologique et la relation entre ce comportement, la variété et le lieu géographique. Nous avons aussi déterminé les isothermes de sorption de ces gommes pour prédire les conditions idéales de leur conservation. Les résultats obtenus à l'issue de cette étude ont démontré clairement la présence des corrélations morphologie, composition chimique et coordonnées géographiques de différents cultivars étudiés. De même, les études structurales et rhéologiques ont montré des différences significatives entre les gommes purifiées. Cette différence semble être liée à une variation du rapport galactose/mannose et de la masse molaire observée selon les variétés. Par ailleurs, les études des isothermes de sorption ont permis de fournir des informations complémentaires sur leurs hygroscopicités et par conséquence sur les conditions idéales de leur conservation
Key message
Arbuscular mycorrhizal symbiosis strengthened the mechanisms developed by carob to withstand drought stress, including improved water relations, increased cell wall rigidity and osmolytes accumulation, and enhanced oxidative stress alleviation.
Abstract
The present investigation was carried out to provide more insight into the influence of arbuscular mycorrhizal fungi (Funneliformis mosseae, Rhizophagus fasciculatus, and Rhizophagus intraradices) on drought tolerance of carob. Non-mycorrhizal (NM) and arbuscular mycorrhizal (AM) carob plants were subjected to two watering regimes, 75% of field capacity (well water) or 25% of field capacity (water stress). Obtained results showed that stressed AM plants exhibited increased performance in terms of growth and biomass production, water and nutrient acquisition, and oxidative stress alleviation compared to NM plants. In fact, under limited water regime, AM plants maintained high stomatal conductance and high relative water content (over 94%) due to their high water and nutrient uptake efficiency. Moreover, AM plants especially those associated with F. mosseae maintained high membrane integrity (over 80%), high cell wall rigidity, and high leaf water potential and osmotic potential at full turgor and at turgor loss, while these parameters steeply decreased in NM plants. Furthermore, drought-stressed AM plants showed decreased hydrogen peroxide and malondialdehyde contents associated with increased activities of superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and catalase compared to their relative NM plants. Thus, AMF strengthened the mechanisms involved in drought tolerance of carob by improving water relations, increasing cell wall rigidity, and enhancing oxidative stress alleviation. Funneliformis mosseae was the most effective in improving carob tolerance to drought stress.
Extraction condition efficiency of total polyphenols was studied on carob pods (Ceratonia siliqua L.) by performing extractions with various solvent systems, in order to evaluate and optimize the conditions of the recovery of polyphenols. Maximum amounts of polyphenols were found in 80% acetone extracts versus water inefficiency. Total polyphenol contents, determined by Folin-Ciocalteu method, were estimated at 11.19 mg/g (dry weight) and the chromatographic analyses demonstrated that gallic acid is the major polyphenol compound of the extracts (45% of polyphenols by dry weight material).
Carob leaves and different parts of fruits were also analyzed for their total polyphenols and gallic acid content. It was found that the mature leaves of the year contain more polyphenols and gallic acid (45.26 mg/g of total polyphenols and 17.01 mg/g of gallic acid). Tegument extracts only contained low amounts of polyphenols and gallic acid (26.30 mg/g of total polyphenols and 2.51 mg/g of gallic acid), while only traces were detected in germ and endosperm (1.33 mg/g and 0.80 mg/g of total polyphenols; 0.99 mg/g and 0.72 mg/g of gallic acid, respectively). It should be noted that there are some additional phenolic compounds present whose structures still need to be determined
The flowering phenology of five carob cultivars (two female, two male, and one hermaphroditic cultivar) was studied at two sites in Tarragona (NE Spain) in 1988 and 1989. Developmental stages from initiation of inflorescence buds to maturity and withering were characterized. Flower number per inflorescence is greatest in male cultivars (25.7–43.5), intermediate in hermaphrodite ones (20.8), and lowest in female cultivars (17.0). In four of the five cultivars, all flowers within an inflorescence mature more or less at the same time, but inflorescences of one male cultivar develop acropetally. Maturity stages (dehiscence and/or receptivity) last 7–10 d. The percentage of flowers aborting before maturity is high in female and hermaphrodite cultivars (17.9 to 36.6%) and low in male cultivars (3.5 to 6.1%). In the study area, blooming starts in mid-August, and lasts until mid-November. Development of inflorescences is more staggered in female and hermaphrodite cultivars than in male ones, so that trees of the former cultivars typically bear inflorescences in as many as six developmental stages, whereas male trees never carry inflorescences of more than four developmental stages. For the same reason, blooming periods of female and hermaphroditic cultivars are longer (two months or more) than with male cultivars (1–1.5 months).
The mineral concentration of flowers and the seasonal fluctuation of macro- [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg)] and micronutrients [iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu)] in leaves of male, female, and hermaphroditic carob trees (Ceratonia siliqua L.) were studied. The nutrient dynamics were linked not only to phenological events, but also to the gender of the trees. The females were able to allocate more nutrients to leaves than male trees, even though male flowers were richer in particular elements such as N and Zn. The hermaphrodites supported the development of both inflorescences and fruits with a lower seasonal variation and a lower leaf nutrient concentration, as compared to the other genders, which may indicate a more efficient use of resources. Flowers had, in general, a higher concentration of N, P, and K, and a lower Mn concentration than in leaves. Flowers of the females had a lower nutrient concentration compared to males or hermaphrodites.
The methanol leaf extracts of female cultivars of the carob tree [Ceratonia siliqua L. (Fabaceae)] and of hermaphrodite and male trees were investigated for their contents of phenolic compounds, their in vitro antioxidant activity, measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging and linoleic acid system assays, and their in vitro tumor growth inhibition on HeLa cells. The different cultivars and trees showed high levels of phenols, and considerable variations in the amount of these compounds. The extracts showed significant radical scavenging activity (RSA), which was not significantly affected by the gender of the tree. From the female cultivars tested, Galhosa exhibited the highest RSA. Gender significantly affected the antioxidant activity of the extracts measured by the linoleic acid system assay, and males and hermaphrodites showed the highest activities. The extracts displayed a remarkable ability to inhibit tumor cell proliferation, and their bioactivity varied with different cultivars or trees tested. Extracts from male and hermaphrodite trees exhibited higher capacity to inhibit the proliferation of HeLa cells than the female cultivars.
This research evaluated the in vitro apoptotic inducing properties of a methanol leaf extract of carob tree (Ceratonia siliqua L.) on a human cervical adenocarcinoma cell line (HeLa). The cell viability effect on a prostate (DU-145), breast (MDA-MB-231) and colon cell line (HCT-166) was also assessed. The effect of the extract on reactive oxygen species (ROS) production by HeLa cells was studied, and a phytochemical evaluation was made by high performance liquid chromatography with diode array detection (HPLC). Total mean yield of identified phenols was 261.1 mg/g DW, and (+)-catechin was the major compound (76.8 mg/g DW). The extract decreased cell viability in a dose-and time-dependent manner, and a more pronounced effect on HeLa line was observed. In vitro cytotoxic activity was associated with apoptosis, but not to the increase of ROS production. Among the tested compounds identified in the extract, the highest activity was detected with gallic acid (GA), (+)-catechin and quercetin, with reductions in HeLa cells viability down to 8.0, 11.9 and 27.1%, respectively. This is the first report on the apoptotic activity of a leaf extract of carob tree on a human cancer cell line, suggesting that it may be a potential source of chemopreventive compounds.
Carob pod, germ, and seed were analyzed for moisture, ash, protein, fat, carbohydrates, and particularly for their tannin content. Recovery of tannins as affected by various solvent extraction systems was investigated. Carob pod meal contained high levels of carbohydrates (45%), appreciable amounts of protein (3%), and low levels of fat (0.6%). Germ and seed meal contained more fat and less carbohydrates compared to the carob pod. Seventy percent acetone was the most effective solvent for the extraction and recovery of tannins. Carob pod contains a mean value of 19 mg of total polyphenols/g, 2.75 mg of condensed tannins (proanthocyanidins)/g, 0.95 mg of hydrolysable tannins (gallo- and ellagitannins)/g. Germ contained higher concentration of total polyphenols (40.8 mg/g) and tannins (16.2 mg of condensed tannins/g and 2.98 mg of hydrolysable tannins/g) while only traces of these compounds were detected in carob seed.
This work aimed to evaluate the phytochemical content and to determine the antioxidant and cytotoxic activities of methanol extracts of the carob tree (Ceratonia siliqua L.) germ flour. The extracts were rich in phenolic compounds, had considerable antioxidant activity, and reduced the viability of cervical (HeLa) cancer cells. The chemical content and the biological activities of the extracts were significantly affected by gender and cultivar. Female cultivar Galhosa had the highest levels of phenolic compounds, and the highest antioxidant activity. Extracts from the hermaphrodite trees and from the female cultivars Galhosa and Costela/Canela exhibited the highest cytotoxic activity. The most abundant compound was theophylline. The phenolic content was correlated to both antioxidant and cytotoxic activities. Our findings provide new knowledge about the health implications of consuming food supplemented with carob germ flour.
Aged red wines possess significantly different polyphenolic composition compared with young ones, mainly due not only to formation of polymeric compounds but also because of oxidation, hydrolysis, and other transformations that may occur in native grape phenolics during aging. Representative Greek, single-variety, aged red wines were examined for total phenol, total flavanol, and total anthocyanin content using spectrophotometry, and attempts were made to establish correlations with the antiradical, reducing, and hydroxyl free radical scavenging activity. In addition, HPLC analyses were carried out, to ascertain whether individual polyphenols are actually responsible for the antioxidant effects of aged red wines. It was found that total flavanols are the class of polyphenols that account for hydroxyl free radical scavenging efficacy and to a lesser extent for antiradical and reducing ability, whereas there was a less significant link between the antioxidant properties and the total phenolics and only a weak relationship to total anthocyanin content. The correlation of the antioxidant properties with the principal polyphenols showed that individual compounds are weakly associated with all the antioxidant parameters, suggesting that the expression of antioxidant activity in aged red wines is rather a consequence of synergism between various phenolics, and it is not simply attributed to specific constituents.
Green tea, mainly through its constituents epigallocatechin gallate, epigallocatechin, epicatechin gallate and epicatechin, has demonstrated anticarcinogenic activity in several animal models, including those for skin, lung and gastro-intestinal tract cancer, although less is known about colorectal cancer. Quercetin, the major flavonoid present in vegetables and fruit, exerts potential anticarcinogenic effects in animal models and cell cultures, but less is known about quercetin glucosides. The objectives of this study were to investigate (i) the antioxidant activity of the phenolic compounds epicatechin, epigallocatechin gallate, gallic acid and quercetin-3-glucoside; (ii) the cytotoxicity of different concentrations of epicatechin, epigallocatechin gallate, and gallic acid; (iii) the cellular uptake of epicatechin, epigallocatechin gallate, gallic acid and quercetin-3-glucoside and (iv) their effect on the cell cycle. Human colon adenocarcinoma cells were used as experimental model. The results of this study indicate that all dietary flavonoids studied (epicatechin, epigallocatechin gallate, gallic acid and quercetin-3-glucoside) show a significant antioxidant effect in a chemical model system, but only epigallocatechin gallate or gallic acid are able to interfere with the cell cycle in Caco2 cell lines. These data suggest that the antioxidant activity of flavonoids is not related to the inhibition of cellular growth. From a structural point of view, the galloyl moiety appears to be required for both the antioxidant and the antiproliferative effects.
British Journal of Cancer (2002) 86, 1645–1651. DOI: 10.1038/sj/bjc/6600295 www.bjcancer.com
© 2002 Cancer Research UK
Water and ethanol crude extracts from black pepper (Piper nigrum) were investigated for their antioxidant and radical scavenging activities in six different assay, namely, total antioxidant activity, reducing power, 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities. Both water extract (WEBP) and ethanol extract (EEBP) of black pepper exhibited strong total antioxidant activity. The 75 microg/ml concentration of WEBP and EEBP showed 95.5% and 93.3% inhibition on peroxidation of linoleic acid emulsion, respectively. On the other hand, at the same concentration, standard antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and alpha-tocopherol exhibited 92.1%, 95.0%, and 70.4% inhibition on peroxidation of linoleic acid emulsion, respectively. Also, total phenolic content in both WEBP and EEBP were determined as gallic acid equivalents. The total phenolics content of water and ethanol extracts were determined by the Folin-Ciocalteu procedure and 54.3 and 42.8 microg gallic acid equivalent of phenols was detected in 1 mg WEBP and EEBP.
Ghrelin is an orexigenic hormone that may affect substrate utilization in humans. Ghrelin is influenced by macronutrients, but the effects of insoluble dietary fiber and polyphenols are unknown. We investigated the effects of a polyphenol-rich insoluble dietary fiber preparation from carob pulp (carob fiber) on postprandial ghrelin responses and substrate utilization. Dose-dependent effects of the consumption of carob fiber were investigated in a randomized, single-blind, crossover study in 20 healthy subjects, aged 22-62 y. Plasma total and acylated ghrelin, triglycerides, and serum insulin and nonesterified fatty acids (NEFA) levels were repeatedly assessed before and after ingestion of an isocaloric standardized liquid meal with 0, 5, 10, or 20 g of carob fiber over a 300-min period. The respiratory quotient (RQ) was determined after consumption of 0 or 20 g of carob fiber. Carob fiber intake lowered acylated ghrelin to 49.1%, triglycerides to 97.2%, and NEFA to 67.2% compared with the control meal (P < 0.001). Total ghrelin and insulin concentrations were not affected by consumption of a carob fiber-enriched liquid meal. Postprandial energy expenditure was increased by 42.3% and RQ was reduced by 99.9% after a liquid meal with carob fiber compared with a control meal (P < 0.001). We showed that the consumption of a carob pulp preparation, an insoluble dietary fiber rich in polyphenols, decreases postprandial responses of acylated ghrelin, triglycerides, and NEFA and alters RQ, suggesting a change toward increased fatty acid oxidation. These results indicate that carob fiber might exert beneficial effects in energy intake and body weight.
The natural antioxidant gallic acid (GA) was isolated from fruits of a medicinal Indonesian plant, Phaleria macrocarpa (Scheff.) Boerl. The structure was identified on the basis of spectroscopic analysis and comparison with authentic compound. GA demonstrated a significant inhibition of cell proliferation in a series of cancer cell lines and induced apoptosis in esophageal cancer cells (TE-2) but not in non-cancerous cells (CHEK-1). Observation of the molecular mechanism of apoptosis showed that GA up-regulated the pro-apoptosis protein, Bax, and induced caspase-cascade activity in cancer cells. On the other hand, GA down-regulated anti-apoptosis proteins such as Bcl-2 and Xiap. In addition, GA also induced down-regulation of the survival Akt/mTOR pathway. In non-cancerous cells, we observed delayed expression of pro-apoptosis related proteins. Our results suggest that GA might be a potential anticancer compound. However, in depth in vivo studies are needed to elucidate the exact mechanism.
The natural antioxidant gallic acid (GA) was isolated from fruits of a medicinal Indonesian plant, Phaleria macrocarpa (Scheff.) Boerl. The structure was identified on the basis of spectroscopic analysis and comparison with authentic compound. GA demonstrated a significant inhibition of cell proliferation in a series of cancer cell lines and induced apoptosis in esophageal cancer cells (TE-2) but not in non-cancerous cells (CHEK-1). Observation of the molecular mechanism of apoptosis showed that GA up-regulated the proapoptosis protein, Bax, and induced caspase-cascade activity in cancer cells. On the other hand, GA down-regulated antiapoptosis proteins such as Bcl-2 and Xiap. In addition, GA also induced down-regulation of the survival Akt/mTOR pathway. In non-cancerous cells, we observed delayed expression of pro-apoptosis related proteins. Our results suggest that GA might be a potential anticancer compound. However, in depth in vivo studies are needed to elucidate the exact mechanism.
Carob bean is the fruit of Ceratonia siliqua L, an evergreen sclerophyllous perennial and long-producing tree, which naturally grows on barren, rocky and dry regions of the Mediterranean basin and other parts of world, with similar climate. Deseeded carob pod (husk), although rich in water-soluble sugars (mainly sucrose), has a very low protein content and high levels of tannins (mainly condensed), which minimize the nutritional value of carob beans. The present review attempts to appraise the current status and future potentials of the carob bean in food as well as feed industry and environment upgrading.
Several phenolic components were extracted from willow samples by using the same extraction procedure, purified, and analyzed by spectrophotometry and gas chromatography. The components analyzed were total phenolics, condensed tannins, and phenolic glycosides. Methods for drying the material, extraction, and analysis are tested and discussed.
The genetic diversity of 15 carob (Ceratonia siliqua L.) cultivars located in an experimental field from Algarve (Portugal) was evaluated over 7 years using 12 fruit and seed phenotypic characters, in order to characterize carob cultivars. The values of morphological traits obtained by cultivar were compared with those from other countries of the Mediterranean basin. Statistically significant differences were found between cultivars for all characters which were examined, what indicates a high genetic diversity. The relationship among these characters was analyzed by principal component analysis (PCA) resulting in the separation of these cultivars classed in four groups (clusters I–IV) and in four ungrouped cultivars. A three dimension of the model was found to be significant and explained 74.5% of the total variation, in which the first component accounting for 34.6% of the total variation is dominated by fruit characters, while the second component is dominated by seed characters. Cultivars plotted on the left-lower quadrant on the space determined by principal components 1 and 2 are characterized by fruits with high seed yield more appropriated for industrial rentability. The correlation analyses established by cultivar provided a specific understanding about the way how fruit and seed characteristics correlate within each cultivar. This approach can be useful for the development of a breeding programme, aiming to increase the seed yield, seed thickness, individual and total seed weight by fruit, characteristics that are determinant to improve the industrial exploitation of carob. # 2007 Published by Elsevier B.V.
Phytochemicals, as plant components with discrete bio-activities towards animal biochemistry and metabolism are being widely examined for their ability to provide health benefits. It is important to establish the scientific rationale to defend their use in foods, as potential nutritionally active ingredients. Phytochemicals could provide health benefits as: (1) substrates for biochemical reactions; (2) cofactors of enzymatic reactions; (3) inhibitors of enzymatic reactions; (4) absorbents/sequestrants that bind to and eliminate undesirable constituents in the intestine; (5) ligands that agonize or antagonize cell surface or intracellular receptors; (6) scavengers of reactive or toxic chemicals; (7) compounds that enhance the absorption and or stability of essential nutrients; (8) selective growth factors for beneficial gastrointestinal bacteria; (9) fermentation substrates for beneficial oral, gastric or intestinal bacteria; and (10) selective inhibitors of deleterious intestinal bacteria. Such phytochemicals include terpenoids, phenolics, alkaloids and fiber. Research supporting beneficial roles for phytochemicals against cancers, coronary heart disease, diabetes, high blood pressure, inflammation, microbial, viral and parasitic infections, psychotic diseases, spasmodic conditions, ulcers, etc is based on chemical mechanisms using in vitro and cell culture systems, various disease states in animals and epidemiology of humans. However, it must be emphasized that a distinction needs to be drawn between the types of information that can be obtained from studies in vitro, in animals and in humans. Mechanisms of action must certainly be established in vitro; however, the efficacy of these same ingredients with their mechanisms of action, must also be demonstrated in vivo. The rapid growth in the use of phytochemicals in nutraceutical and functional foods requires that the food and pharmaceutical industries face new challenges: in addressing worldwide public concern over the efficacy and safety of supplements and foods claimed to be health-promoting; in government regulations related to safety, labeling and health claims for products that contain phytochemicals; in the manufacturing of foods with different qualities and stabilities; and in marketing issues, particularly as they relate to consumers' recognizing added value.© 2000 Society of Chemical Industry
The aim of this study was to assess the antioxidant value and antiproliferative activity of some vegetables such as raw garlic
(Allium sativum L), white and yellow, and red onions (Allium cepa L), red and green peppers (Capsicum annuum L.), and white cabbage (Brassica oleracea var. capitata f. alba), which were harvested in the same year and in the same geographical and climatic conditions. It was found that the highest
content of some bioactive compounds (25.19±2.2mg GAE/g, 3.84±0.3mg CE/g, 4.88±0.3mg CE/g, 59.20±0.8μg CE/g, 1992±99.8μg
AA/g, 452.7±26.1mg CGE/kg DW for polyphenols, flavonoids, tannins, flavanols, ascorbic acid, and anthocyanins, respectively)
and the antioxidant activity (41.32±3.9, 31.05±2.7, 59.17±5.2 and 58.94±5.1μM TE/g for DPPH, FRAP, CUPRAC and ABTS,
respectively) was in red onion. Methanol extracts in concentration of 1,000μgml−1 of garlic and red onion exhibited antiproliferative activity (83.1±2.1 and 85.0±3.2% of viability, respectively). In
spite of relatively high antioxidant activity in methanol extracts of yellow onion, red and green pepper, no antiproliferative
activity on both tumor cell lines was registered. In conclusion, among the studied vegetables raw red onion was the preferable.
The interrelationship was in the following order: red onion>white onion=yellow onion>red pepper>garlic=green pepper>white
cabbage. The antiproliferative activities of these vegetables were different: some samples reacted only on Calu-6 and the
others—on SNU-601. Thus, vegetables from the same cultivation place were examined for their antioxidant and antiproliferative
activities with four different methods. Based on obtained data a direct comparison between these vegetables was possible for
the first time.
The antiradical activities of various antioxidants were determined using the free radical, 2,2-Diphenyl-1-picrylhydrazyl (DPPH*). In its radical form. DPPH* has an absorption band at 515 nm which dissappears upon reduction by an antiradical compound. Twenty compounds were reacted with the DPPH* and shown to follow one of three possible reaction kinetic types. Ascorbic acid, isoascorbic acid and isoeugenol reacted quickly with the DPPH* reaching a steady state immediately. Rosmarinic acid and δ-tocopherol reacted a little slower and reached a steady state within 30 min. The remaining compounds reacted more progressively with the DPPH* reaching a steady state from 1 to 6 h. Caffeic acid, gentisic acid and gallic acid showed the highest antiradical activities with a stoichiometry of 4 to 6 reduced DPPH* molecules per molecule of antioxidant. Vanillin, phenol, γ-resorcylic acid and vanillic acid were found to be poor antiradical compounds. The stoichiometry for the other 13 phenolic compounds varied from one to three reduced DPPH* molecules per molecule of antioxidant. Possible mechanisms are proposed to explain the experimental results.
The aerial parts of two endemic Pimpinella [Pimpinella anisetum Boiss. & Ball. and Pimpinella flabellifolia (Boiss.) Benth. ex Drude] were hydro-distilled to produce oils in the yields of 2.07% (v/w) and 2.61% (v/w), respectively. The oils were analysed by GC and GC/MS. Twenty-one and nineteen components were identified, representing 99.5% and 99.7% of the oils, respectively. The main compounds of P. anisetum were (E)-anethole (82.8%) and methyl chavicol (14.5%), whereas limonene (47.0%), (E)-anethole (37.9%) and α-pinene (6.0%) were the major constituents of P. flabellifolia. The oils were screened for their possible antioxidant activities by two complementary test systems, namely DPPH free radical-scavenging and β-carotene/linoleic acid systems. In the first case, P. anisetum oil exerted greater antioxidant activity than that of P. flabellifolia oil with an IC50 value of 5.62 ± 1.34 μg/ml. In the β-carotene/linoleic acid test system, the oil of P. anisetum was superior to P. flabellifolia with 70.5% ± 2.86 inhibition rate. Essential oils of the plants studied here were also screened for their antimicrobial activities against six bacteria and two fungi. The oils showed moderate antimicrobial activity against all microorganisms tested.
Red meat intake is associated with an increased risk of developing cancer. This is possibly related to the heme content of red meat. Plant derived polyphenols might protect from cancer development via their antioxidant activities. In this study, the impact of an aqueous extract of carob (CE) on hemin-modulated proliferation was investigated. CE, gallic acid (GA) and a known iron chelator (deferoxamine: DFO) significantly reduced the number of human colon cancer HT29 cells. CE and GA were more effective under serum-free conditions than in normal cell culture medium. These effects were abolished by addition of 1 μM hemin at low concentrations of CE and GA. At higher concentrations of CE and GA, both substances reduced cell number despite hemin supplementation. Effects of CE, GA and DFO on cell number could not be linked to iron chelation even though CE and DFO were capable of chelating iron. Furthermore, the effects of high CE concentration point to antioxidative effects other than iron chelation. However, a connection to a reduction of colorectal cancer risk due to consumption of meat with high heme content by CE could not be drawn, since the effective concentrations are beyond the physiologically relevant concentrations.
The literature addresses dietary fiber (DF) and antioxidants (AOX) separately as nonrelated compounds. This paper proposes to show that DF and AOX could be approached jointly in nutrition and health studies because around 50% of the total dietary antioxidants, mainly polyphenolics, traverse the small intestine linked to dietary fiber. These antioxidants have received little attention so far. They release the fiber matrix in the colon by the action of the bacterial microbiota, producing metabolites and an antioxidant environment. The content of polyphenols associated with DF in different foods and their potential health-related properties, including animal experiments and human trials, are reviewed. It is concluded that the transportation of dietary antioxidants through the gastrointestinal tract may be an essential function of DF.
Phenolic ingredients of an aqueous carob extract are well characterized and consist of mainly gallic acid (GA). In order to assess possible chemopreventive mechanisms of carob, which can be used as a cacao substitute, effects on expression of genes related to stress response and drug metabolism were studied using human colon cell lines of different transformation state (LT97 and HT29). Stress-related genes, namely catalase (CAT) and superoxide dismutase (SOD2), were induced by carob extract and GA in LT97 adenoma, but not in HT29 carcinoma cells. Although corresponding protein products and enzyme activities were not elevated, pretreatment with carob extract and GA for 24 h reduced DNA damage in cells challenged with hydrogen peroxide (H(2)O(2)). In conclusion, carob extract and its major phenolic ingredient GA modulate gene expression and protect colon adenoma cells from genotoxic impact of H(2)O(2). Upregulation of stress-response genes could not be related to functional consequences.
Infants aged 3-21 months with acute diarrhea of bacterial and viral origin were treated as inpatients with oral rehydration fluid and randomly received for up to 6 days either a tannin-rich carob pod powder (40% tannins or 21.2% polyphenols and 26.4% dietary fiber), 1.5 g/kg/day (n = 21) to a maximum of 15 g, or an equivalent placebo (n = 20). The duration of the diarrhea from admission was 2.0 +/- 0.27 days in the test group and 3.75 +/- 0.30 days in the placebo group (p less than 0.001). Normalized defecation, body temperature, and weight and cessation of vomiting were reached more quickly by the patients who received the test substance. The test substance was well accepted and tolerated.