HPLC and GC Characterization of Dicliptera bupleuroides Aerial Parts and Evaluation of Its Anti-Inflammatory Potential in Vitro, in silico and in Vivo Using Carrageenan and Formalin Induced Inflammation in Rat Models.

The current study aimed to evaluate the anti-inflammatory activity of Dicliptera bupleuroides Nees aerial parts methanol extract and its different fractions namely hexane, chloroform, ethyl acetate and butanol in vitro using cyclooxygenase inhibitory assay (COX-2). In vivo anti-inflammatory evaluation was performed using carrageenan and formalin induced inflammation in rat models followed by molecular docking. High performance liquid chromatography (HPLC) and gas chromatography coupled with mass chromatography (GC/MS) analyses were used for chemical analyses of the tested samples. The tested samples showed significant inhibition in COX-2 inhibitory assay where methanol extract (DBM) showed the highest inhibitory potential at 100 µg/mL estimated by 67.86 %. At a dose of 400 mg/kg, all of the examined samples showed pronounced results in carrageenan induced acute inflammation in rat model at 4th h interval with DBM showed the highest efficiency displaying 65.32 % inhibition as compared to the untreated rats. Formalin model was employed for seven days and DBM exhibited 65.33 % and 69.39 % inhibition at 200 and 400 mg/kg, respectively approaching that of the standard on the 7th day. HPLC revealed the presence of caffeic acid, gallic acid and sinapic acid, quercetin and myricetin in DBM. GC/MS analysis of its hexane fraction revealed the presence of 16 compounds belonging mainly to fatty acids and sterols that account for 85.26 % of the total detected compounds. Molecular docking showed that hexadecanoic acid followed by decanedioic acid and isopropyl myristate showed the best fitting within cyclooxygenase-II (COX-II) while nonacosane followed by hexatriacontane and isopropyl myristate revealed the most pronounced fitting within the 5-lipoxygenase (5-LOX) active sites. Absorption, metabolism, distribution and excretion and toxicity prediction (ADMET/ TOPKAT) concluded that most of the detected compounds showed reasonable pharmacokinetic, pharmacodynamic and toxicity properties that could be further modified to be more suitable for incorporation in pharmaceutical dosage forms combating inflammation and its undesirable consequences.

Anti-Inflammatory and Phytochemical Analysis of the Crude Leaves Extracts of Boscia Coriacea Graells and Uvaria Leptocladon Oliv.

Background: The objective of this study was to evaluate the anti-inflammatory activities and phytochemical composition of the leaves extracts of Boscia coriacea Graells and Uvaria leptocladon Oliv. Methods: The powdered leaves of Boscia coriacea Graells and Uvaria leptocladon Oliv were extracted by maceration and soxhlet extraction methods. Anti-inflammatory activity of the leaves extracts of Boscia coriacea Graells and Uvaria leptocladon Oliv were evaluated using carrageenan-induced paw edema model. Standard methods were used for analysis of phytochemical composition of the leaves extracts of Boscia coriacea Graells and Uvaria leptocladon Oliv. Data analysis was done using one way analysis of variance. Results: U. leptocladon Oliv (200 mg/kg) and B. coriacea Graells (200 mg/kg) showed percent inhibition on mice paw edema of 86% and 75% after six hours of carrageenan injection, respectively. The ethanol fraction (100 mg/kg) of U. leptocladon Oliv showed the highest anti-inflammatory effect after six hours of carrageenan injection. The phytochemical analysis of the leaves extracts of B. coriacea and U. leptocladon revealed the presence of tannins, alkaloids, cardiac glycoside, flavonoids, phenols, quinones, and saponins. Conclusion: The crude leaves extracts of B. coriacea Graells and U. leptocladon Oliv contain phytochemicals with anti-inflammatory activities.

Ultrasound-Assisted Water Extraction of Mastocarpus stellatus Carrageenan with Adequate Mechanical and Antiproliferative Properties.

Ultrasound-assisted water extraction was optimized to recover gelling biopolymers and antioxidant compounds from Mastocarpus stellatus. A set of experiments following a Box-Behnken design was proposed to study the influence of extraction time, solid liquid ratio, and ultrasound amplitude on the yield, sulfate content, and thermo-rheological properties (viscoelasticity and gelling temperature) of the carrageenan fraction, as well as the composition (protein and phenolic content) and antiradical capacity of the soluble extracts. Operating at 80 °C and 80 kHz, the models predicted a compromise optimum extraction conditions at ~35 min, solid liquid ratio of ~2 g/100 g, and ultrasound amplitude of ~79%. Under these conditions, 40.3% carrageenan yield was attained and this product presented 46% sulfate and good mechanical properties, a viscoelastic modulus of 741.4 Pa, with the lowest gelling temperatures of 39.4 °C. The carrageenans also exhibited promising antiproliferative properties on selected human cancer cellular lines, A-549, A-2780, HeLa 229, and HT-29 with EC50 under 51.9 µg/mL. The dried soluble extract contained 20.4 mg protein/g, 11.3 mg gallic acid eq/g, and the antiradical potency was equivalent to 59 mg Trolox/g.

The impact of ι- and κ-carrageenan addition on freezing process and ice crystals structure of strawberry sorbet frozen by various methods.

The aim of this work was to study the influence of ι- and κ-carrageenan addition to strawberry sorbet prepared by various freezing methods (cryostat and conventional freezer) on freezing kinetics and ice crystals structure. Four variants of strawberry-based sorbet were prepared: with no additive, with 0.1% addition of ι- and κ-carrageenan, and 1:1 blend of both. Freezing curves and freezing time, as well as the size and morphology of ice crystals, were analyzed. The addition of carrageenan stabilizers resulted in freezing process elongation. Reference samples were characterized by different dynamics of the freezing process. The addition of carrageenan increased the final temperature of the samples, compared to the reference sample (even 2.6 °C of difference). Additives reduced ice crystals growth. The most efficient was mixture of both fractions of carrageenan--the samples in this variant frozen in conventional freezer were characterized by the smallest ice crystals diameter--average 5.44 µm, while for the reference sample it was 17.79 µm. PRACTICAL APPLICATION: This research can give a new path for the carrageenan's hydrolysates application--not only for dairy products. Iota carrageenan fraction was recommended for dairy products according to its water-holding capacity in the presence of calcium ions. This study showed that mix of ι- and κ-carrageenan could bring positive results to such product as sorbet without any milk proteins addition. This experiment data could also be helpful when optimizing frozen desserts production in small gastronomy--especially the temperature and time of freezing modifications.

Addendum to Weiner, M.L. (2016) Parameters and Pitfalls to Consider in the Conduct of Food Additive Research, Carrageenan as a Case Study. Food Chemical Toxicology 87, 31-44.

This paper is an addendum to a 2016 paper outlining pitfalls and parameters to consider in the conduct of food additive research with carrageenan (Fd. Chem. Tox. 87, 31-44 (2016)). The literature on the food additive, "carrageenan," contains many publications which either erroneously misuse the name, carrageenan, for a sample which is not carrageenan, but "degraded carrageenan" or "poligeenan" and also conduct studies without understanding the physical/chemical properties of carrageenan. Degraded carrageenan and poligeenan are not food additives and have a completely different physical/chemical and toxicological properties from carrageenan. Two recent publication examples, one in vivo and one in vitro, demonstrate the serious misunderstanding promulgated by incorrect sample identity/purity and poor study conduct. These new publication examples reiterate the problems in the literature summarized by the Weiner (2016). It is important to have thorough, rigorous peer review of all studies using carrageenan in vivo or in vitro.

Optimization of a gelled emulsion intended to supply ω-3 fatty acids into meat products by means of response surface methodology.

The optimization of a gelled oil-in-water emulsion was performed for use as fat replacer in the formulation of ω-3 PUFA-enriched cooked meat products. The linseed oil content, carrageenan concentration and surfactant-oil ratio were properly combined in a surface response design for maximizing the hardness and minimizing the syneresis of the PUFA delivery system. The optimal formulation resulted in a gelled emulsion containing 40% of oil and 1.5% of carrageenan, keeping a surfactant-oil ratio of 0.003. The gel was applied as a partial fat replacer in a Bologna-type sausage and compared to the use of an O/W emulsion also enriched in ω-3. Both experimental sausages contributed with higher ω-3 PUFA content than the control. No sensory differences were found among formulations. The selected optimized gelled oil-in-water emulsion was demonstrated to be a suitable lipophilic delivery system for ω-3 PUFA compounds and applicable in food formulations as fat replacer.

Preliminary assessment of carrageenan as excipient for extrusion/spheronisation.

The current study pursues the suitability of different types of carrageenan as a novel extrusion aid. The aim was to find out a suitable substitution to the commonly used microcrystalline cellulose (MCC). The types of kappa-carrageenan were found to be the most appropriate material and the required fraction to produce acceptable pellets in the formulation was determined. The investigation showed that 5% of kappa-carageenan was necessary to produce pellets without MCC. Similar formulations produced with MCC or kappa-carrageenan were compared with respect to size and shape of the pellets. kappa-Carrageenan required higher water content for the formation of pellets, but the formulation was more robust as the optimal range of water content was much broader. Hence, kappa-Carrageenan seems to be a suitable and promising extrusion aid. The study showed that the substitution of MCC by kappa-carrageenan in formulations is possible and the produced pellets were of high quality.