The botanical species Ceratonia siliqua L., commonly referred to as the Carob tree, and locally as “L’Kharrûb”, holds significance as an agro-sylvo-pastoral species, and is traditionally utilized in Morocco for treating a variety of ailments. This current investigation aims to ascertain the antioxidant, antimicrobial, and cytotoxic properties of the ethanolic extract of C. siliqua leaves (CSEE). Initially, we analyzed the chemical composition of CSEE through high-performance liquid chromatography with Diode-Array Detection (HPLC-DAD). Subsequently, we conducted various assessments, including DPPH scavenging capacity, β-carotene bleaching assay, ABTS scavenging, and total antioxidant capacity assays to evaluate the antioxidant activity of the extract. In this study, we investigated the antimicrobial properties of CSEE against five bacterial strains (two gram-positive, Staphylococcus aureus, and Enterococcus faecalis; and three gram-negative bacteria, Escherichia coli, Escherichia vekanda, and Pseudomonas aeruginosa) and two fungi (Candida albicans, and Geotrichum candidum). Additionally, we evaluated the cytotoxicity of CSEE on three human breast cancer cell lines (MCF-7, MDA-MB-231, and MDA-MB-436) and assessed the potential genotoxicity of the extract using the comet assay. Through HPLC-DAD analysis, we determined that phenolic acids and flavonoids were the primary constituents of the CSEE extract. The results of the DPPH test indicated a potent scavenging capacity of the extract with an IC₅₀ of 302.78 ± 7.55 µg/mL, which was comparable to that of ascorbic acid with an IC₅₀ of 260.24 ± 6.45 µg/mL. Similarly, the β-carotene test demonstrated an IC₅₀ of 352.06 ± 12.16 µg/mL, signifying the extract’s potential to inhibit oxidative damage. The ABTS assay revealed IC₅₀ values of 48.13 ± 3.66 TE µmol/mL, indicating a strong ability of CSEE to scavenge ABTS radicals, and the TAC assay demonstrated an IC₅₀ value of 165 ± 7.66 µg AAE/mg. The results suggest that the CSEE extract had potent antioxidant activity. Regarding its antimicrobial activity, the CSEE extract was effective against all five tested bacterial strains, indicating its broad-spectrum antibacterial properties. However, it only showed moderate activity against the two tested fungal strains, suggesting it may not be as effective against fungi. The CSEE exhibited a noteworthy dose-dependent inhibitory activity against all the tested tumor cell lines in vitro. The extract did not induce DNA damage at the concentrations of 6.25, 12.5, 25, and 50 µg/mL, as assessed by the comet assay. However, the 100 µg/mL concentration of CSEE resulted in a significant genotoxic effect compared to the negative control. A computational analysis was conducted to determine the physicochemical and pharmacokinetic characteristics of the constituent molecules present in the extract. The Prediction of Activity Spectra of Substances (PASS) test was employed to forecast the potential biological activities of these molecules. Additionally, the toxicity of the molecules was evaluated using the Protox II webserver.

Carob (Ceratonia siliqua L.) is a tree species native to the Mediterranean region and belongs to the Fabaceae family. The tree is well-known for its sweet and nutritious fruits, which have been used for long time as a nutritious food. In addition to the edible fruits, the carob tree also produces seeds that are highly prized for their ability to produce carob gum (locust bean gum). The carob seed consists of three main components: the shell, the endosperm, and the embryo. The shell is the outermost layer of the seed, followed by the endosperm, which is the largest part of the seed and contains high levels of carbohydrates and proteins. The embryo is the smallest part of the seed and is rich on bioactive compounds. Carob seed constituents have attracted considerable attention due to their exceptional nutritional and therapeutic properties in various industries, including food, medicine, pharmaceuticals, cosmetics, and textiles. The high content of bioactive compounds in carob seeds, such as polyphenols, tannins, and flavonoids, is believed to be responsible for their antioxidant and anti-inflammatory properties. The use of carob seed constituents in the food industry is mainly due to their ability to act as thickeners and stabilizers in various foods. They are used as a substitute for other thickening agents such as guar gum and carrageenan, due to their superior properties. In the pharmaceutical industry, carob seeds have been found to have antidiabetic, antihyperlipidemic, and anticancer properties, among others. The cosmetics industry is also interested in the ingredients of carob seed, as they can improve hydration and elasticity of the skin. They are also used as a natural alternative to synthetic thickeners in cosmetic formulations. The textile industry has also recognized the potential of carob seed constituents, as they can be used as a natural dye and as a sizing agent to improve the strength and durability of textiles. In summary, carob seed constituents offer a wide range of applications in various industries, owing to their high content of bioactive compounds, excellent nutritional and therapeutic profile, and ability to act as thickeners, stabilizers, and antioxidants. This review has highlighted the latest findings on the chemical composition, applications, and health benefits of carob seed constituents.

Ceratonia siliqua L. is a medicinal plant that has long been used in traditional Moroccan medicine to treat many diseases. This study aimed to assess the impact of the stages of the immature phase of carob pulp (M1, M2, M3, M4, and M5) on phytochemical composition, antioxidant activity, and antidiabetic activity of Ceratonia siliqua L. The identification of the phenolic profile by HPLC-UV/MS-MS and the study of the antidiabetic effect by in silico, in vitro, and in vivo studies were carried out for extracts with high contents of phenolic compounds from immature wild carob pulp from the communes of Timoulit (TM), Bin Elouidane (AW), and Ouaouizerth (TG) in the province of Azilal in the Béni Mellal-Khénifra region. The results revealed a gradual increase in total sugar content over the pulp’s ripening period, reaching a value of 2134 ± 56.23 mg GE/100 g fresh weight (FW) for TG. The three locations showed peak values for total polyphenol content (TPC), total flavonoid content (TFC), and total condensed tannin (TCT) at the M2 stage. AW had the highest concentrations of TPC (3819 ± 226.4 mg GAE/100 g FM), TFC (1034 ± 57.08 mg QE/100 g FM), and TCT (1472 ± 28.46 mg CE/100 g FM). The DPPH assay (7892 ± 296.1 mg TE/100 g FM) and the FRAP assay (278.2 ± 7.85 mg TE/100 g FM) both demonstrated that the TG zone is a highly potent antioxidant zone. In contrast, the AW site exhibited a markedly elevated value of 725.4 ± 103.6 mg TE/100 g FM in the ABTS assay. HPLC-UV-MS/MS analysis showed that the methanolic extracts of immature carob pulp (MEICP) from the three areas contained several different chemical compounds. The most prevalent were 3-O-p-coumaroyl-5-O-caffeoylquinic acid, quercetin 3-methyl ether, gallic acid, and galloylquinic acid. Immature carob pulp extract (ICPE) from AW showed the strongest in vitro inhibition of pancreatic α-amylase (IC₅₀ = 0.405 µg/mL) and TG extracts were most potent against intestinal α-glucosidase (IC₅₀ = 0.063 µg/mL). In vivo, AW, TG, and TM extracts significantly reduced postprandial glycemia in rats, with AW having the greatest effect. These results highlight the antidiabetic potential of ICPE. The 3-O-p-Coumaroyl-5-O-caffeoylquinic acid showed better affinity for α-amylase compared to acarbose and interacted significantly with several amino acid residues of the enzyme. Similarly, this molecule and 3,4-Dicaffeoylquinic acid demonstrated a strong affinity for α-glucosidase, suggesting their potential as natural inhibitors of enzymes involved in carbohydrate metabolism. Most of the compounds are not substrates of P-glycoprotein and exhibited high intestinal absorption. Furthermore, the majority of these compounds did not act as inhibitors or substrates of CYP450 enzymes, reinforcing their suitability for development as oral medications. These results underscore the potential of immature carob pulp as a promising antidiabetic agent.

The significance of conducting research for its application has been noted as a result of the rising global food production and waste generation. As a result, there is increasing interest in fruits and vegetable seeds that contain bioactive chemicals, such as those that are obtained from orange seeds. In the current work, orange seed powder replaced wheat flour at 0, 2.5, 5, 7.5, and 10% levels, to observe changes in physicochemical features of developed biscuits. Proximate analysis of orange seed powder and wheat flour revealed that orange seed powder has high fat, fiber, protein, and ash contents as compared to wheat flour, whereas moisture contents in wheat flour were high. In developed biscuits, the highest values (percentage) of ash ( ), fiber ( ), protein ( ), and fat ( ) were found in biscuits developed with 10% orange seed powder. Orange seed powder was a comparatively good source of both macro and micro minerals, as compared to wheat flour. High contents of selenium ( ), iron ( ), zinc ( ), and manganese ( ) mg/100 g, present in orange seed powder, were the prominent findings of this research work, as wheat flours were observed to be deficient in these trace minerals. Contents of calcium, magnesium, potassium, zinc, manganese, zinc, and selenium in control biscuits were found , , , , ,