# Phytocomplex Influences Antimicrobial and Health Properties of Concentrated Glycerine Macerates
## 3 Discussion
Furthermore, the findings obtained by IM-SPME-GC-MS are very considerable because, thanks to this methodology, they highlight for the first time a different volatile chemical composition of the samples analyzed. As shown in the results, components belonging to the terpene family, among which terpinen-4-ol, terpinolene, spathulenol and caryophyllene oxide, were only found in RN extracts. These volatile compounds define the tropism of RN C-GMs for inflammatory diseases of the respiratory tract. Moreover, components belonging to the family of fatty acids have been identified in the C-GMs of FC, CB and AG, even if samples extracted from the same botanical species showed different concentrations between the two sources. Fatty acids are known to be a primary energy substrate, as well as structural components of cell membranes, second messengers, anti-inflammatory and anti-microbial compounds that have beneficial effects for the health of themother and her offspring, for cardiovascular health, insulin sensitivity, metabolic syndrome, cancer, critically ill patients, and immune system disorders [21; 22; 23]. It is generally acknowledged that oxidative stress can play a crucial role in the development or chronicity of respiratory tract diseases. In this context, the use of natural products able to neutralize the reactive oxygen species can be a valuable aid in healing [24; 25]. The DPPH analysis shows that the B-C-GMs are active at lower concentrations than A-C-GMs, and that C-GMs of CB and AG have the best antioxidant action. Bacterial or viral microbial agents can determine several cooling conditions; among bacterial ones, that associated with _S. pyogenes_ is one of the most frequent causes of pharyngitis. The microbial analysis shows that both sources of AG, in equal measure, are the most active C-GMs against this strain as they are able to inhibit bacterial growth at concentrations lower than 2.5% _v/v_, and to suppress it at concentrations equal to 5% _v/v_ and 10% _v/v_ for AG C-GM from A and B producer, respectively. At these concentrations, this C-GM exerts a selective action towards the pathogenic strain without altering the viability of the probiotic strains. This selective action is already known for natural compounds, and our data support what has recently been observed by [PERSON] et al. [26]. Therefore, the in vitro data support the integration of ingestible gargles made with C-GMs to treat oral bacterial affections that potentially require the use of antibiotics. Furthermore, a microbial infection induces a natural inflammatory response mediated by the immune system, which is able to counteract the infection but, at the same time, generates important discomforts such as pain, fever and impaired tissue functions. The analyses carried out with the major anti-inflammatory bud-derived GMs, when tested on PBMCs grown in the presence of pro-inflammatory stimulus (LPS), show that the most concentrated extract (RN-B) can significantly counteract the pro-inflammatory response induced by the LPS, bringing the ratio of pro- and anti-inflammatory cytokines to levels close to unstimulated PBMCs (_p_ < 0.05). This data supports the traditional use of C-MG of RN as an immunomodulatory agent of the inflammatory response. Finally, the data obtained from the toxicity test indicate that the doses traditionally used (5 gtt of C-GMs in some water/day) have not cytotoxic effects. Our in vitro data, in addition to supporting the traditional integration of CB, AG and RN C-GMs in the diet of people with diseases of the upper respiratory tract, highlight how the quality of C-GMs correlates with their activity, and the importance of standardizing all practices carried out before the maceration step in order to obtain more concentrated products which are, therefore, active at lower concentrations.