Increased anti-inflammatory activity and enhanced phytochemical concentrations in superfine powders obtained by controlled differential sieving process from four medicinal plants.

Anti-inflammatory effect of Rosa canina, Salix alba, Scrophularia nodosa and Hedera helix were studied in LPS-stimulated primary peripheral blood mononuclear cells (PBMCs) from mice (n=18) by comparing homogeneous powders of small microparticles (50-100 µm, 100-180 µm and 180-315 µm) obtained from plants via a controlled differential sieving process (CDSp) versus total plant materials obtained via hydroethanolic (HE) extraction. Further, phytochemical composition of the fine powders and HE extracts was determined by LC-PDA-ESI/MS analyses. Results showed that a one-hour pretreatment of PBMCs with fine powders, particularly those with superfine particle sizes (i.e. 50-100 µm and 100-180 µm), significantly inhibited TNFα, IL-1ß, IL-6 and NO production in LPS-stimulated PBMCs, by at least ca. 20% more than HE extracts (all, p<0.05). For each of the plants studied, their superfine powdered fractions were more concentrated in phenolic contents than their HE extracts. Overall, our results further confirm CDSp, as an environmentally friendly method, for improving the concentration of bioactive compounds as well as their biological activities.

Phytochemicals, Antioxidant Attributes and Larvicidal Activity of Mercurialis annua L. (Euphorbiaceae) Leaf Extracts against Tribolium confusum (Du Val) Larvae (Coleoptera; Tenebrionidae).

This study reports the link between larvicidal activity and the phytochemical composition of male and female leaf extracts of Mercurialis annua L. (M. annua) from four Tunisian regions: Bizerte, Jandouba, Nabeul and Beja. Their antioxidant activity was evaluated using DPPH (2, 2-diphenyl-1-picrylhydrazyl) assays. Phenolic compounds were identified and quantified using liquid chromatography coupled with a UV detector and mass spectrometry (LC-UV-ESI/MS). Higher antioxidant activity (AOA) was found in the leaves of male plant extracts than of female ones. The leaves of male and female plant extracts from Bizerte exhibited the highest AOA: 22.04 and 22.78 mg Trolox equivalent/g dry matter (mg TE/g DM), respectively. For both sexes, plant extracts from Beja had the lowest AOA with 19.71 and 19.67 mg TE/g DM for male and female plants, respectively. Some phenolic compounds such as narcissin, gallocatechin, rutin, epigallocatechin and epicatechin were identified and quantified using LC-MS, which highlighted the abundance of narcissin and rutin in the male leaves of M. annua. We noted that the interaction between the sex of plants and the provenance had a significant effect on TFC (F = 6.63; p = 0.004) and AOA (F = 6.53; p = 0.004) assays, but there was no interaction between sex and origins for TPC (F = 1.76; p = 0.19). The larvicidal activity of aqueous leaf extracts of M. annua against Tribolium confusum (Du Val) (T. confusum), an insect pest of flour and cereal seeds, showed that the mortality could reach 100% after 48 h of exposure in the Bizerte region. The LC50 values for the leaf extract were low in Bizerte, with 0.003 and 0.009 g/mL for male and female plants, respectively, succeeded by Jandouba, which displayed 0.006 and 0.024 g/mL for male and female plants, respectively. Nabeul showed 0.025 g/mL for male plants and 0.046 g/mL for female plants and Beja showed 0.037 and 0.072 g/mL for male and female plants, respectively. This is the first time that a study has revealed a negative correlation between the antioxidant activity and the larvicidal activity of the leaf extracts of M. annua with the following correlation coefficients of Perason: r = -0.975 and r = -0.760 for male and female plants, respectively.

Effect of controlled differential sieving processing on micronutrient contents and in vivo antioxidant activities of Hibiscus sabdariffa L. calyxes powder.

Sun-dried calyxes of Hibiscus sabdariffa were finely grinded and fractionated by controlled differential sieving processing (CDSp) into four granulometric classes. The obtained powder fractions were examined for their micronutrient contents and in vivo antioxidant properties in high-fat diet induced oxidation model rats. Unsieved powder and lyophilized ethanolic extract were used for comparison. Micronutrient contents and antioxidant properties were significantly influenced (p < 0.05) by particle sizes. The mineral, vitamin, carotenoid and anthocyanin contents were shown to be higher in smaller powder particles. Comparatively, the ethanolic extract powder had the higher carotenoid, vitamin and anthocyanin contents while the superfine powders < 180 µm and 180-212 µm had the higher mineral contents and antioxidant properties which was characterized by high superoxide dismutase, catalase activities and low malondialdehyde production. Plant grinding followed by CDSp may be an option to improve micronutrient content and antioxidant activity of plant powders.

Micronutrients and in vivo antioxidant properties of powder fractions and ethanolic extract of Dichrostachys glomerata Forssk. fruits.

Dichrostachys glomerata powders were processed by sieve fractionation and ethanolic extraction followed by freeze-drying. The micronutrient contents and the in vivo antioxidant properties of powder fractions in rats' high-fat diet-induced oxidation model were determined. Sieved fractionation was achieved by finely grinding the Dichrostachys fruits and fractionating on a sieve column to retain particle of sizes <180, 180-212, 212-315, and ≥315 µm. Unsieved powder and lyophilized ethanolic extract were used as control. All the powders were examined for the minerals, total carotenoids, and vitamins contents. For in vivo antioxidant properties assessment, the individual powder was dissolved in water and administered to rats at the dose of 250 mg/kg body weight. Oxidation was induced by treating the rat with high-fat diet, and the measured parameters were malondialdehyde, superoxide dismutase, and catalase activities. The results showed a significant influence (p < .05) of particle size on the micronutrient contents and in vivo antioxidant properties. The smaller the particle size of the powder fractions, the higher the minerals, vitamins, total carotenoids contents, and antioxidant properties. Comparatively, the ethanolic powder had the highest carotenoids content, while the powders of particle size <180 µm and 180-212 µm had the highest minerals and vitamin contents. The highest antioxidant properties were characterized by high superoxide dismutase, catalase activities, and low malondialdehyde production. The grinding of Dichrostachys fruit followed by controlled differential sieving process may compete with ethanol extraction for an efficient concentration of bioactive compounds and micronutrients except carotenoids.

Optimisation of phytochemical characteristics and antioxidative properties of Foeniculum vulgare Mill. seeds and Ocimum basilicum L. leaves superfine powders using new parting process.

INTRODUCTION: Fennel (Foeniculum vulgare Mill.) and basil (Ocimum basilicum L.) are two aromatic medicinal plants. Recently, a new parting process which is named CDS "Comminution and to control Diffraction Sieving" is taken into consideration and its positive effect on the extraction of bioactive compounds from the plants, without any solvent, is reported. OBJECTIVE: Study the effect of CDS on phytochemical properties of superfine powders of fennel seeds and basil leaves. METHODS: Fennel seeds and basil leaves superfine powders were fractionated as follows: 100-180 µm, 180-315 µm, 315-500 µm, > 500 µm and unsieved superfine powders. Extraction of polyphenols was carried out using hydromethanolic maceration. The essential oils were extracted by maceration with dichloromethane. The antioxidant activities were evaluated by DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Using liquid chromatography ultraviolet electrospray ionisation mass spectrometry (LC-UV-ESI-MS), concentrations of characterised phenolic compounds were measured. The essential oils compositions were characterised using gas chromatography mass spectrometry (GC-MS). RESULTS: In fennel seeds, the 100-180 µm fraction had maximum antioxidant activity and LC-UV-ESI-MS analyses proved that the best extraction of certain polyphenols was obtained in the 100-180 µm fraction. In basil leaves, the best antioxidant activity corresponded to the 315-500 µm fraction and LC-UV-ESI-MS analyses showed that the polyphenols were concentrated in the < 315 µm fractions. For both plants, GC-MS presented that the essential oils were concentrated in the 315-500 µm fraction. CONCLUSION: CDS as a new green parting process leads to improve the phytochemical properties of these two superfine plant powders in the specific granulometric classes.

Powders with small microparticle size from Hedera helix and Scrophularia nodosa exhibited high preventive antioxidant activity against H2O2-induced oxidative stress in mouse primary spleen cells.

The purpose of this study was to examine the effects of powder particle size on the cytoprotective and antioxidant activity of Hedera helix (HH) and Scrophularia nodosa (SN), two medicinal plants more commonly known as ivy and figwort, against H2O2-induced oxidative stress in mouse primary spleen cells. Thus, the preventive effects of powders of 3 different granulometric classes (50-100 µm, 100-180 µm and 180-315 µm) and those of the hydroethanolic (HE) extract from HH and SN on oxidative stress were compared by monitoring reactive oxygen species (ROS) formation, malondialdehyde (MDA) production, and the activity of enzymatic antioxidants including catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx). Results showed that pretreatment with the 3 fine powders from both plants generally offered to H2O2-exposed spleen cells, a protection against oxidative stress, highlighted by a significant decrease of both ROS formation and the level of MDA (p < 0.001), and a significant increase of GPx activity (p < 0.05). The two superfine powders (i.e. 50-100 µm and 100-180 µm), at 250 µg/mL, were more effective in modulating all oxidative stress markers studied than both HE extracts (p < 0.01), and the powder with the highest particular size (i.e., 180-315 µm) (p < 0.01). Compared to untreated cells, our results suggest that pretreatment with powders, in particular the superfine fractions, has relatively restored the levels of antioxidant-related enzymes including GPx, CAT and SOD. In summary, our results suggest differential effects between the 3 different fine powders studied, with the best cytoprotective and antioxidant activities being in favor of the superfine powders.

Improvement of cytoprotective and antioxidant activity of Rosa canina L. and Salix alba L. by controlled differential sieving process against H2O2-induced oxidative stress in mouse primary splenocytes.

In this study, cytoprotective and antioxidant activities of Rosa canina (RC) and Salix alba (SA), medicinal plants, were studied on mouse primary splenocytes by comparing Controlled Differential Sieving process (CDSp), which is a novel green solvent-free process, versus a conventional technique, employing hydroethanolic extraction (HEE). Thus, preventive antioxidant activity of three plant powders of homogeneous particle sizes, 50-100 µm, 100-180 µm and 180-315 µm, dissolved directly in the cellular buffer, were compared to those of hydroethanolic (HE) extract, at 2 concentrations (250 and 500 µg/mL) in H2O2-treated spleen cells. Overall, compared to HE extract, the superfine powders, i. e., fractions < 180 µm, at the lowest concentration, resulted in greater reactive oxygen species (ROS) elimination, increased glutathione peroxidase (GPx) activity and lower malondialdehyde (MDA) production. Better antioxidant and preventive effects in pre-treated cells were found with the superfine powders for SA (i. e., 50-100 µm and 100-180 µm, both p < 0.001), and with the intermediate powder for RC (i. e., 100-180 µm, p < 0.05) versus HE extract. The activity levels of catalase (CAT) and superoxide dismutase (SOD) in pretreated splenocytes exposed to H2O2, albeit reduced, were near to those in unexposed cells, suggesting that pretreatment with the fine powders has relatively restored the normal levels of antioxidant-related enzymes. These findings supported that CDSp improved the biological activities of plants, avoiding the use of organic solvents and thus it could be a good alternative to conventional extraction techniques.