Maria Bellumori

Storage method, drying processes and extraction procedures strongly affect the phenolic fraction of rosemary leaves: an HPLC/DAD/MS study.

The Rosmarinus officinalis L. is widely known for its numerous applications in the food field but also for the increasing interest in its pharmaceutical properties. Two groups of compounds are mainly responsible for the biological activities of the plant: the volatile fraction and the phenolic constituents. The latter group is mainly constituted by rosmarinic acid, by a flavonoidic fraction and by some diterpenoid compounds structurally derived from the carnosic acid. The aim of our work was to optimize the extractive and analytical procedure for the determination of all the phenolic constituents. Moreover the chemical stability of the main phenols, depending on the storage condition, the different drying procedures and the extraction solvent, have been evaluated. This method allowed to detect up to 29 different constituents at the same time in a relatively short time. The described procedure has the advantage to being able to detect and quantify several classes of compounds, among them numerous minor flavonoids, thus contributing to improving knowledge of the plant. The findings from this study have demonstrated that storing the raw fresh material in the freezer is not appropriate for rosemary, mainly due to the rapid disappearing of the rosmarinic acid during the freezing/thawing process. Regarding the flavonoidic fraction, consistent decrements, were highlighted in the dried samples at room temperature if compared with the fresh leaf. Rosmarinic acid, appeared very sensitive also to mild drying processes. The total diterpenoidic content undergoes to little changes when the leaves are freeze dried or frozen and limited losses are observed working on dried leaves at room temperature. Nevertheless it can be taken in account that this fraction is very sensitive to the water presence during the extraction that favors the conversion of carnosic acid toward it oxidized form carnosol. From our findings, it appear evident that when evaluating the phenolic content in rosemary leaves, several factors, mainly the type of storage, the drying process and the extraction methods, should be carefully taken into account because they can induce partial losses of the antioxidant components.

Rosmarinic acid prevents lipid peroxidation and increase in acetylcholinesterase activity in brain of streptozotocin-induced diabetic rats.

We investigated the efficacy of rosmarinic acid (RA) in preventing lipid peroxidation and increased activity of acetylcholinesterase (AChE) in the brain of streptozotocin‐induced diabetic rats. The animals were divided into six groups (n = 8): control, ethanol, RA 10 mg/kg, diabetic, diabetic/ethanol and diabetic/RA 10 mg/kg. After 21 days of treatment with RA, the cerebral structures (striatum, cortex and hippocampus) were removed for experimental assays. The results demonstrated that the treatment with RA (10 mg/kg) significantly reduced the level of lipid peroxidation in hippocampus (28%), cortex (38%) and striatum (47%) of diabetic rats when compared with the control. In addition, it was found that hyperglycaemia caused significant increased in the activity of AChE in hippocampus (58%), cortex (46%) and striatum (30%) in comparison with the control. On the other hand, the treatment with RA reversed this effect to the level of control after 3 weeks. In conclusion, the present findings showed that treatment with RA prevents the lipid peroxidation and consequently the increase in AChE activity in diabetic rats, demonstrating that this compound can modulate cholinergic neurotransmission and prevent damage oxidative in brain in the diabetic state. Thus, we can suggest that RA could be a promising compound in the complementary therapy in diabetes. Copyright

An innovative approach to the recovery of phenolic compounds and volatile terpenes from the same fresh foliar sample of Rosmarinus officinalis L.

Rosmarinus officinalis L. is a plant of relevant commercial interest because of its volatile fraction and also its phenolic constituents which are both well known for their numerous properties. Nevertheless, an extractive method suitable to recovering both the aromatic and phenolic fractions from the same fresh foliar tissue has not yet been reported. In this work we have optimized a two-step procedure able to recover first the phenolic compounds and successively the volatile terpenes from the same foliar sample. The recovery of the whole phenolic fraction, partially degraded using a traditional extractive method, was guaranteed and we observed a significant increment in the amount of volatile terpenes compared to a traditional extraction procedure. We also highlight crucial information on the enzymatic activity of the endogenous oxidases that rapidly transform the phenolic substrates, mainly the rosmarinic acid. Our results suggest that this extractive procedure could also be used for other aromatic plants, thus providing a useful tool for more complete analyses of the main phytochemicals available in fresh foliar samples and creating the possibility of incrementing yields of volatile compounds.

An in vitro evaluation of the antibacterial activity of the non-volatile phenolic fraction from rosemary leaves.

In this study, the aim was to evaluate the antimicrobial action of the non-volatile phenols of rosemary leaves against two Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and two Gram-positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis). Three extracts with different phenolic compositions were tested. By the agar disc diffusion method, Gram-positive bacteria were more sensitive to the extracts, and S. epidermidis showed the highest inhibition zones. Overall, all the extracts tested by the broth dilution method showed higher activity than results from the agar disc diffusion method. The minimal bactericidal concentration values indicated that E. coli was the most susceptible strain. This study demonstrated that the flavonoidic fraction of rosemary leaves does not play a crucial role as antimicrobial agents against these microorganisms. The most active extract was characterised by the highest amount of non-volatile terpenoidic compounds.

Anti-neuropathic effects of Rosmarinus officinalis L. terpenoid fraction: relevance of nicotinic receptors.

Traditional uses and current results highlight the neuroprotective properties of Rosmarinus officinalis L. The compelling need for novel strategies able to relieve neuropathic pain encouraged us to analyze different rosemary leaf extracts in rats following chronic constriction injury (CCI) of sciatic nerve. Ethanol, acetone, and the innovative ultrasound-hexane extractive methods were used to obtain: EE, AE, and for hexane extracts UREprel and URE. Extracts were characterized in terms of typical constituents and repeatedly administered to CCI-rats (13-days treatment, from the day of surgery). URE showed the best efficacy and potency in reducing hypersensitivity to noxious- and non-noxious stimuli and spontaneous pain. URE contained the higher quantity of the terpenoid carnosic acid (CA) and its efficacy was compared to pure CA. Histological analysis of the sciatic nerve revealed that URE prevented axon and myelin derangement, edema and inflammatory infiltrate. In the dorsal horn of the spinal cord, URE did not reduce astrocyte activation. Both the pain reliever and the neuroconservative effects of URE were significantly prevented by the nicotinic receptor (nAChR) antagonist mecamylamine. In conclusion, the hexane-ultrasound rosemary extract is able to reduce neuropathic hypersensitivity and protect nervous tissues. Effectiveness is mainly related to the terpenoid fraction by mechanisms involving nAChRs.

The antimicrobial effects of three phenolic extracts from Rosmarinus officinalis L., Vitis vinifera L. and Polygonum cuspidatum L. on food pathogens

Abstract In this study three phenolic extracts were examined, without volatile fraction, against common food pathogens. The samples, all suitable for food application, were from the leaves of Rosmarinus officinalis L., Vitis vinifera L., and the root of Polygonum cuspidatum L. The microorganisms tested were Escherichia coli O157:H7, Salmonella Enteritidis, Salmonella Typhi, Yersinia enterocolitica and Listeria monocytogenes, well-known as important food pathogens. The results demonstrated a microbicidal activity of all the tested compounds at different concentrations; the rosemary extract showed greater efficacy than the other compounds against the tested microorganisms. In particular, the best results were obtained with rosemary extract against E. coli O157:H7 and L. monocytogenes with values of 200 and 270 μg/mL, respectively. Our results show that rosemary extract, often present as a natural antioxidant in food, can also be proposed as a natural disinfectant in the food field.

Effects of ionizing radiation on bio-active plant extracts useful for preventing oxidative damages

Abstract Humans are exposed to ionizing radiations in medical radiodiagnosis and radiotherapy that cause oxidative damages and degenerative diseases. Airplane pilots, and even more astronauts, are exposed to a variety of potentially harmful factors, including cosmic radiations. Among the phytochemicals, phenols are particularly efficient in countering the oxidative stress. In the present study, different extracts obtained from plant food, plant by-products and dietary supplements, have been compared for their antioxidant properties before and after irradiation of 140 cGy, a dose absorbed during a hypothetical stay of three years in the space. All the dry extracts, characterized in terms of vitamin C and phenolic content, remained chemically unaltered and maintained their antioxidant capability after irradiation. Our results suggest the potential use of these extracts as nutraceuticals to protect humans from oxidative damages, even when these extracts must be stored in an environment exposed to cosmic radiations as in a space station.

Characterization and comparison of cold brew and cold drip coffee extraction methods: Cold brew and cold drip coffee extraction methods

BACKGROUND Each region of the world has its own methods, protocols, instruments and procedures regarding how to brew coffee. The final result in the cup is strongly affected by the extraction method, and many studies have focused on this subject. However, few studies have investigated slow, cold extraction methods, despite their popularity among baristas. Therefore, the present study aimed to characterize and compare two cold extraction methods: cold brew and cold drip. RESULTS Physical and chemical analyses were used to describe coffee beverages in terms of pH, total solids, refractive index, density and viscosity. Caffeine and cinnamic acids were quantified using high-performance liquid chromatography (HPLC)/diode array detector and HPLC/mass spectrometry. A sensory evaluation included aroma, flavor and textural attributes. CONCLUSIONS Significant differences were found in the chemical and physical parameters, both between and within the two methods, as a function of the extraction temperature and contact time. Similarly, the sensory evaluation found differences in flavor profiles, as measured in terms of bitterness, sweetness, sourness and global intensity.

What kind of coffee do you drink? An investigation on effects of eight different extraction methods

The chemical composition of brewed coffee depends on numerous factors: the beans, post-harvest processing and, finally, the extraction method. In recent decades, numerous coffee-based beverages, obtained using different extraction techniques have entered the market. This study characterizes and compares eight extraction coffee methods from a chemical-physical point of view, starting from the same raw material. Specifically, three types of Espresso, Moka, French Press, and 3 filter coffee that for the first time are reported in the scientific literature Cold Brew, V60, and Aeropress are compared. Physical measurements included the quantification of total dissolved solids, density, pH, conductivity, and viscosity. Chemical analyses identified 15 chlorogenic acids (CGAs): six caffeoylquinic acids, one p-Coumaroylquinic acid, one Feruloylquinic Acid, four Caffeoylquinic lactones, and three Dicaffeoylquinic acids. Maximum caffeine and CGA concentrations were found in Espresso coffees, while Moka and filtered coffees were three to six times less concentrated. The classic Espresso method was most efficient for caffeine and CGA recovery, with a yield almost double that of other methods. Per-cup caffeine and CGAs were higher in Cold Brew than Espresso coffees, as a function of the volume of beverage, which ranged from 30 mL (for espresso) to 120 mL (for filtered coffees). In light of these results, it is not possible to establish how many cups of coffee can be consumed per day without exceeding the recommended doses, since according to the applied brewing method, the content of the bioactive substances varies considerably.

Recovery and stability over time of phenolic fractions by an industrial filtration system of olive mill wastewaters: a three years study

BACKGROUND The recovery of phenolic compounds from olive milling is recognized as strategic for producers. The aim of this work was to evaluate the quality and stability of retentates obtained from olive mill wastewaters treated with a membrane filtration system constituted by a micro-, ultra- and nanofiltration, followed by a final reverse osmosis, over three crop seasons. Efficiency was evaluated in terms of phenolic amount in the retentates and of organic load in the final discarded waters. Phenolic compounds were quantified using tyrosol as external standard. RESULTS Our study highlighted a reproducibility of the process over years and a low organic load in permeates from reverse osmosis. Hydroxytyrosol was very stable in the liquid products at 18-28 °C over 24 months of storage. The retentates from reverse osmosis showed the highest phenolic content (78.6 mg g-1 dry matter in 2015), associated with a potassium content of 22 g kg-1 . CONCLUSIONS The liquid concentrated retentates showed an unexpected stability over time of their bioactive phenolic compounds, particularly of hydroxytyrosol. These samples recovered from olive mill wastewaters can be good sources of natural antioxidants and potassium to guarantee the correct intake and to formulate new food ingredients or food products.