The 42nd Symposium Chromatographic Methods of Investigating Organic Compounds : Book of abstracts

The 42nd Symposium Chromatographic Methods of Investigating Organic Compounds : Book of abstracts. June 4-7, 2019, Szczyrk, Polan

Phytochemical and Pharmacological Evaluation of the Residue By-Product Developed from the Ocimum americanum (Lamiaceae) Postdistillation Waste

The yield of essential oils in plants is not high and postdistillation wastes rich in phenolic compounds could be used to enhance the profitability of essential oil-bearing plants. The aim of the study was to evaluate polyphenols in a dry extract obtained from the postdistillation waste of the American basil (Ocimum americanum L.) herb, and to conduct the screening of its pharmacological activities. Rosmarinic acid, caffeic acid and rutin were identified in the extract using high-performance thin-layer chromatography. The high-performance liquid chromatography analysis found the presence of a plethora of polyphenols in the extract. Rosmarinic acid, luteolin-7-O-glucoside and rutin were as the main compounds. The total phenolic content in the extract was 106.31 mg GAE/g and free radical scavenging activity against 2,2-diphenyl-1-picrylhydrazyl evaluated as IC50 was 0.298 mg/mL. The tested extract dose-dependently decreased the paw edema in rats, suggesting its potent anti-inflammatory property. The acute toxicity study indicates its safety. Thus, the O. americanum hydrodistilled residue by-product is the promising source of biologically active compounds with significant antioxidant and anti-inflammatory effects

Polish Yellow Sweet Clover (Melilotus officinalis L.) Honey, Chromatographic Fingerprints, and Chemical Markers

A case study of Polish Melilotus officinalis honey was presented for the first time. Gas chromatography–mass spectrometry (GC-MS) (after steam distillation, Soxhlet extraction, ultrasonic solvent extraction, and solid phase extraction (SPE)) and targeted high performance liquid chromatography with a photodiode array detector (HPLC-PAD) were applied to determine the characteristic components of honey. While ubiquitous in most honeys, carbohydrates, terpene derivatives, and phenylacetic acid dominated in the Soxhlet extracts (25.54%) and in the application of SPE (13.04%). In addition, lumichrome (1.85%) was found, and may be considered as a marker of this honey. Due to the presence of these compounds, Polish yellow sweet clover honey is similar to French lavender honeys. The major compounds determined in the methanolic extract were (+)-catechine (39.7%) and gallic acid (up to 30%), which can be regarded as specific chemical markers of the botanical origin of melilot honey. With respect to total phenolic and flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays were determined spectrophotometrically. The honey exhibited a moderate antioxidant activity, typical for light honeys, which correlates well with its phenolic and flavonoid composition

Macrofungi as a source of bioactive substances

From ancient times medicine and natural products have been closely linked through the use of traditional medicines and natural toxins. Macrofungi have an established history of use especially in traditional oriental medicine. Nowadays, mushrooms comprise a vast and yet largely untapped source of powerful new pharmaceutical products. In particular, and most importantly for modern medicine, they are an unlimited source of compounds that exhibit multidirectional activity: antioxidant, immuno-enhancing, immunostimulatory, antibacterial, antiviral, anti- inflammatory, antidiabetic, antiallergenic and anticarcinogenic [1, 2, 5-8]. In the presented work, a substances with biological activity from mainly Basidiomycota were reviewed, including low-molecular-weight (terpenoids, polyphenols steroids, quinones, amines, cerebrosides) (Tab.1) and high-molecular- weight compounds (homo and heteroglucans, glycans, glycoproteins, glycopeptides, peptides, proteins) (Tab. 2). Selected species of toxic and hallucinogenic mushrooms were also included in the review. At present, mushrooms are considered to be functional food. Beneficial health effects may be achieved through the direct consumption of fruiting bodies or the use of dietary supplements in the form of ready-to-use preparations containing mushroom extracts. The last years is also a period of a flourishing new field of medicine - mycopharmacology. The scientific approach to compounds contained in mushrooms allowed the isolation and identification of many valuable active substances which are used in the prevention and treatment of lifestyle diseases

Polish Yellow Sweet Clover (Melilotus officinalis L.) Honey, Chromatographic Fingerprints, and Chemical Markers

A case study of Polish Melilotus officinalis honey was presented for the first time. Gas chromatography–mass spectrometry (GC-MS) (after steam distillation, Soxhlet extraction, ultrasonic solvent extraction, and solid phase extraction (SPE)) and targeted high performance liquid chromatography with a photodiode array detector (HPLC-PAD) were applied to determine the characteristic components of honey. While ubiquitous in most honeys, carbohydrates, terpene derivatives, and phenylacetic acid dominated in the Soxhlet extracts (25.54%) and in the application of SPE (13.04%). In addition, lumichrome (1.85%) was found, and may be considered as a marker of this honey. Due to the presence of these compounds, Polish yellow sweet clover honey is similar to French lavender honeys. The major compounds determined in the methanolic extract were (+)-catechine (39.7%) and gallic acid (up to 30%), which can be regarded as specific chemical markers of the botanical origin of melilot honey. With respect to total phenolic and flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays were determined spectrophotometrically. The honey exhibited a moderate antioxidant activity, typical for light honeys, which correlates well with its phenolic and flavonoid composition

HPTLC Fingerprinting—Rapid Method for the Differentiation of Honeys of Different Botanical Origin Based on the Composition of the Lipophilic Fractions

Bee honey possess various nutritional and medicinal functions, which are the result of its diverse chemical composition. The numerous bioactive compounds in honey come from flower nectar; thus, the identification of the specific chemical profiles of honey samples is of great importance. The lipophilic compounds from eight monofloral honeys (rape, buckwheat, clover, willow, milk thistle, dandelion, raspberry and sweet yellow clover) were investigated. Analyses of the lipophilic fractions obtained by UAE (ultrasound assisted extraction) and SPE (solid phase extraction) extractions were performed using high-performance thin layer chromatography (HPTLC). Chromatographic and cluster analyses allowed the identification of a unique, colorful pattern of separated compounds with specific Rf values on the HPTLC plate for each type of monofloral honey. HPTLC is a simple and effective visual method of analysis, and it can serve as a basis for authenticating different types of honey

Chemical Profiling of Polyfloral Belgian Honey: Ellagic Acid and Pinocembrin as Antioxidants and Chemical Markers

Chemical profiling of northern Belgian polyfloral honeys was performed to analyse their phenolic compound content (flavonoids and phenolic acids). First, samples were subjected to a standard analysis of their physicochemical properties, and then, the phenolic fraction was isolated and analysed using a HPLC/PAD method. All of the tested honeys showed a common and specific phenolic profile that could be the basis for the differentiation of polyfloral honeys of the Antwerp region from other polyfloral honeys. Chromatographic data indicated a high content of ellagic acid (9.13–13.66 mg/100 g honey), as well as the flavonoid pinocembrin (1.60–1.85 mg/100 g honey) in these honeys. Ellagic acid, a compound with well-defined prohealth activities, might be used as a chemical marker for these honeys. With respect to total phenolic and flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays were determined spectrophotometrically. The honey exhibited a moderate antioxidant activity, typical for light honeys

Spectrophotometric and Chromatographic Assessment of Total Polyphenol and Flavonoid Content in <i>Rhododendron tomentosum</i> Extracts and Their Antioxidant and Antimicrobial Activity

In the literature, the chemical composition of Rhododendron tomentosum is mainly represented by the study of isoprenoid compounds of essential oil. In contrast, the study of the content of flavonoids will contribute to the expansion of pharmacological action and the use of the medicinal plant for medical purposes. The paper deals with the technology of extracts from Rh. tomentosum shoots using ethanol of various concentrations and purified water as an extractant. Extracts from Rh. tomentosum were obtained by a modified method that combined the effects of ultrasound and temperature to maximize the extraction of biologically active substances from the raw material. Using the method of high-performance thin-layer chromatography in a system with solvents ethyl acetate/formic acid/water (15:1:1), the following substances have been separated and identified in all the extracts obtained: rutin, hyperoside, quercetin, and chlorogenic acid. The total polyphenol content (TPC) and total flavonoid content (TFC) were estimated using spectrophotometric methods involving the Folin-Ciocalteu (F-C) reagent and the complexation reaction with aluminum chloride, respectively. A correlation analysis was conducted between antioxidant activity and the polyphenolic substance content. Following the DPPH assay, regression analysis shows that phenolic compounds contribute to about 80% (r2 = 0.8028, p Rh. tomentosum. The extract of Rh. tomentosum obtained by ethanol 30% inhibits the growth of test cultures of microorganisms in 1:1 and 1:2 dilutions of the clinical strains #211 Staphylococcus aureus and #222 Enterococcus spp. and the reference strain Pseudomonas aeruginosa ATCC 10145

Research on Acute Toxicity and the Behavioral Effects of Methanolic Extract from Psilocybin Mushrooms and Psilocin in Mice

The pharmacological activities and acute toxicity of the psilocin (PC) and dried residues of the crude extracts of psychotropic mushrooms were investigated in mice. The hallucinogenic substances were effectively isolated, by using methanol, from the species of Psilocybe semilanceata and Pholiotina cyanopus, that were collected in the north-east region of Poland. The chemical analysis of these extracts, which was performed by liquid chromatography with mass spectrometry detection (LC-MS), indicated the presence of psilocin and other hallucinogenic substances, including indolealkylamines and their phosphorylated analogues. When the pure psilocin or fungal extracts were used, slight differences in determined LD50 values were observed. However, the application of PC evoked the highest level of toxicity (293.07 mg/kg) compared to the activity of extracts from Ph. cyanopus and P. semilanceata, where the level of LD50 was 316.87 mg/kg and 324.37 mg/kg, respectively. Furthermore, the behavioral test, which considered the head-twitching response (HTR), was used to assess the effects of the studied psychotropic factors on the serotonergic system. Both, the fungal extracts and psilocin evoked characteristic serotoninergic effects depending on the dose administered to mice, acting as an agonist/partial agonist on the serotonergic system. A dose of 200 mg/kg 5-hydroxytryptophan (5-HTP) induced spontaneous head-twitching in mice (100% effect), as a result of the formation of 5-hydroxytryptamine (5-HT) in the brain. Compared to the activity of 5-HTP, the intraperitoneal administration of 1mg/kg of psilocin or hallucinogenic extracts of studied mushrooms (Ph. cyanopus and P. semilanceata) reduced the number of head-twitch responses of about 46% and 30%, respectively. In contrast, the administration of PC exhibited a reduction of about 60% in HTR numbers