Metabolomic Profiling and Biological Activities of Pleurotus columbinus Quél. Cultivated on Different Agri-Food Byproducts

The genus Pleurotus (Fr.) P. Kumm (Pleurotaceae, Basidiomycota) comprises a cosmopolitan group of mushrooms highly appreciated for their nutritional value and health-promoting benefits. Despite there being many studies about the phytochemical composition of Pleurotus spp., there are very few reports dealing with the phytochemistry, antioxidant and antimicrobial activities of P. columbinus Quél. In this study, a mass spectrometry ultra-performance liquid chromatography mass spectrometry (UHPLC)-QTOF method, coupled with principal component analysis (PCA), was applied to the P. columbinus metabolome in order to investigate the influence of different agri-food residues as growth substrates for P. columbinus cultivation, on the bioactive chemical profile of fruiting bodies and evaluated their potential as antioxidants and antimicrobials. Additionally, a quantitative HPLC-DAD-MS analysis was conducted on phenolic and flavonoid compounds, that could explain, albeit partially, the observed biological effects of P. columbinus extracts. The qualitative metabolic profile identified 97 metabolites, whereas the quantitative HPLC-DAD-MS analysis confirmed the presence of phenolic and flavonoids, in the mushroom extracts, which also showed intrinsic scavenging/reducing and antimicrobial effects. The antibacterial effects were particularly evident against Escherichia coli, whereas Tricophyton and Aspergillus were the dermatophytes more sensitive to the mushroom extracts. The present study supports more in-depth investigations, aimed at evaluating the influence of growth substrate on P. columbinus antimicrobial and antioxidant properties. The extracts from P. columbinus revealed valuable sources of primary and secondary metabolites, thus suggesting potential applications in the formulation of food supplements with biological properties, above all in terms of antioxidant and antimicrobial properties.

Metformin and sodium dichloroacetate effects on proliferation, apoptosis, and metabolic activity tested alone and in combination in a canine prostate and a bladder cancer cell line

An important approach in tumor therapy is combining substances with different action mechanisms aiming to enhance the antineoplastic effect, decrease the therapeutic dosage, and avoid resistance mechanisms. Moreover, evaluating compounds already approved for the treatment of non-neoplastic diseases is promising for new antineoplastic therapies. Sodium dichloroacetate (DCA) reactivates oxidative phosphorylation in the cancer cell mitochondria, reducing apoptosis resistance in cancer cells. Furthermore, metformin inhibits the proliferation of tumor cells and CD133+ cancer -stem-like cells. In the present study, we evaluated the independent and synergistic effect of metformin and DCA on the metabolic activity, cell proliferation, and apoptosis of a canine prostate adenocarcinoma (Adcarc1258) and a transitional cell carcinoma cell line (TCC1506) in comparison to a primary canine fibroblast culture. Determining metformin uptake in tumor cells was performed by quantitative HPLC. Depending on the dosage, metformin as a single agent inhibited the metabolic activity and cell proliferation of the tumor cells, showing only minor effects on the fibroblasts. Furthermore, 1 mM metformin increased apoptosis over 96 h in the tumor cell lines but not in fibroblasts. Additionally, metformin uptake into the tumor cells in vitro was measurable by quantitative HPLC. Synergistic effects for the combination therapy were observed in both neoplastic cell lines as well as in the fibroblasts. Based on these results, metformin might be a promising therapeutic agent for canine urogenital tumors. Further studies on kinetics, toxicology, bioavailability, and application of metformin in dogs are necessary.

Comparison of Secondary Metabolite Contents and Metabolic Profiles of Six Lycoris Species

Quantitative HPLC analysis was performed on six different species of Lycoris herbs to investigate variation in phytochemical content, especially galantamine and phenylpropanoid-derived compounds. The contents of these compounds differed widely among the Lycoris species, with L. radiata and L. chinensis containing the lowest and highest galantamine contents, respectively. Specifically, the galantamine content of L. radiata was 62.5% higher than that of L. chinensis. Following L. radiata, L. sanguinea contained the next highest galantamine content, which was 59.1% higher than that of L. chinensis. Furthermore, a total of 12 phenylpropanoid-derived compounds were found in the different Lycoris species, where L. sanguinea, L. squamigera, and L. uydoensis had the largest accumulation of these compounds. The total phenylpropanoid content of L. sanguinea was the highest, while that of L. radiata was the lowest. Seven of the phenylpropanoid-derived compounds, rutin, quercetin, catechin, epicatechin gallate, chlorogenic acid, benzoic acid, and kaempferol, were dominant. L. sanguinea, L. uydoensis, and L. squamigera showed amounts of these seven compounds that were 5–6 times greater than those of the other species in the study. To the best of our knowledge, our results provide the most detailed phytochemical information on these species to date, which is valuable for future applications using these medicinal plants.

Establishing pinostrobin reference standard for quantitative analysis of the rhizomes of Boesenbergia pandurata

Boesenbergia pandurata Roxb. Schlecht. (Zingiberaceae), called ``Ngai bun'' in Viet Nam, is one of the Southeast Asian medicinal plants and its rhizomes are used primarily as a spice. This is a perennial, short-stemmed plant, formed by leaf sheaths and can grow up to 50 cm. The leaves are 7-11 cm wide and 25-50 cm long. Its rhizome surfaces are light brown in color, the inner rhizome is yellow, oval-shaped, and has a very aromatic odor. In folklore, Boesenbergia pandurata rhizomes are used as a spice for food processing. This plant contains pinostrobin as the major constituents. Previously showed that pinostrobin compound is the main ingredient together with a variety of biological activities such as antibacterial, inhibition of free radicals, ... Pinostrobin is necessary composition for the screening, testing, and quality evaluation of the rhizomes of B. pandurata species and others in the Zingiberaceae family. This research had conducted a reference standard of pinostrobin isolated from the rhizomes of B. pandurata had 99.26 % purity, which is reliable in medicinal testing. An HPLC method for pinostrobin determination was conducted and The quantitative HPLC analysis was validated for system suitability, selectivity, linearity ranges, and precision. Application of the process to investigate the preparation of extract shown that reflux extraction with ethanol obtained the highest pinostrobin content with 22.05 % in extracts and 2.89 % in dried rhizomes of B. pandurata.

A Simple and Rapid Method for Quantitative HPLC MS/MS Determination of Selected Perfluorocarboxylic Acids and Perfluorosulfonates in Human Serum

Per- and polyfluorinated alkyl substances (PFASs) are ubiquitous environmental contaminants, present in the environment and in the human body. They have raised global concern because of their diffusion in the environment, particularly in water, causing cases of human overexposure due to consumption of contaminated drinking water. Human biomonitoring is the most effective way to characterize human exposure to PFASs, and it is important that as many labs as possible could easily perform this kind of analysis. Analytical methods for quantitation of PFAS mixtures in human serum have been developed, but most of them required materials that are not always easily available in all the laboratories. This paper describes a very simple and accessible HPLC MS/MS method of analysis and quantification of 13 perfluorocarboxylic acids and perfluorosulfonate compounds (belonging to the class of per- and polyfluorinated alkyl substances (PFASs)) in human serum. Method development data provide detailed descriptions of the optimization process in terms of sample preparation, laboratory analysis of human serum samples, determination of analytes by HPLC MS/MS, and describing the pump gradient time, working conditions, and acquisition.

Qualitative and quantitative HPLC-ELSD-ESI-MS analysis of steroidal saponins in fenugreek seed

Fenugreek seeds are known as a source of various compounds, the most common of which are steroidal saponins. However, despite the growing interest in this plant material as a healing agent, spice and dietary supplement ingredient, the composition of Polish fenugreek seeds remains unknown. Therefore, the steroidal saponin complex in the seeds of T. foenum-graecum cultivated in Poland was qualitatively and quantitatively analyzed by the HPLC-ELSDESI-MS method. Two C-18 columns connected in series were used for the first time in analysis of fenugreek saponins and ELS detector parameters were optimized. A total of 26 furostanol saponins were revealed, of which 24 were tentatively identified. The HPLC-ELSD method developed for quantitative analysis was preliminarily validated and the determined amount of steroidal saponins in Polish fenugreek seeds was 0.14 %.