Phenological and Environmental Factors' Impact on Secondary Metabolites in Medicinal Plant Cotinus coggygria Scop.

Cotinus coggygria Scop. (smoketree) is a phytotherapeutically valuable shrub growing in specific areas in many Eurasian countries. Exploring the intrinsic and extrinsic (abiotic) factors that modulate its secondary metabolism has fundamental and applicative importance. Three smoketree plants from the same population were studied for a period of 4.5 months. Their extracts were characterized using LC-MS/MS, HPLC-UV-VIS-DAD and colorimetric assays to determine the chemical composition and antioxidant potential. Multivariate analysis was applied to correlate the metabolomic data with registered habitat variables and phenological stages. The identified and quantified compounds belonged to the flavonoids (myricetin-3-O-galactoside, myricitrin) and hydrolysable tannins groups (pentagalloyl glucose, methyl gallate, methyl digallate I). Phenolic compounds and tannins were synthesized abundantly in the flowering and fruit stages, whereas flavonoids and triterpenes accumulated during senescence. The antioxidant activities varied between detection methods, samplings and individuals and were only punctually correlated with the compound contents in certain phenological stages. Based on the HCAbp analysis, the samples clustered under four groups, according to their metabolic profile. The CCA analysis revealed that during the reproductive stages (flower, fruit or seed), the secondary metabolism of the plants' leaves is sensitive to the action of abiotic factors, while in senescence, the metabolic content is according to the phenological phase. This study provides a first attempt at understanding the interplay between the habitat and the metabolome of smoketree.

Optimization of an Ultrasound-Assisted Extraction Method for the Analysis of Major Anthocyanin Content in Erica australis Flowers.

Erica australis plants have been used in infusions and folk medicine for years for its diuretic and antiseptic properties and even for the treatment of infections. In addition, a recently published thorough study on this species has demonstrated its antioxidant, antibiotic, anti-inflammatory, anticarcinogenic and even antitumoral activities. These properties have been associated with the high content of anthocyanins in E. australis leaves and flowers. The aim of the present research is to optimize an ultrasound-assisted extraction methodology for the recovery of the anthocyanins present in E. australis flowers. For that purpose, a Box Behnken design with response surface methodology was employed, and the influence of four variables at different values was determined: namely, the composition of the extraction solvents (0-50% MeOH in water), the pH level of those solvents (3-7), the extraction temperature (10-70 °C), and the sample:solvent ratio (0.5 g:10 mL-0.5 g:20 mL). UHPLC-UV-vis has been employed to quantify the two major anthocyanins detected in the samples. The extraction optimum conditions for 0.5 g samples were: 20 mL of solvent (50% MeOH:H2O) at 5 pH, with a 15 min extraction time at 70 °C. A precision study was performed and the intra-day and inter-day relative standard deviations (RSDs) obtained were 3.31% and 3.52%, respectively. The developed methodology has been successfully applied to other Erica species to validate the suitability of the method for anthocyanin extraction.

The Study of Antioxidant Components in Grape Seeds.

The paper deals with the study of antioxidant properties of extracts from vine seeds (Vitis vinifera L.) using spectrometric and chromatographic techniques. Ten vine varieties (Cerason, Laurot, Kofranka, Gewürztraminer, Hibernal, Blaufrankisch, Zweigeltrebe, Erilon, Palava, and Welschriesling) obtained from the years 2015, 2016, and 2017 were selected for the study. The antioxidant activity was determined spectrophotometrically using four fundamentally different methods; the content of total polyphenolic compounds was determined using the Folin-Ciocalteu method. In 2015, the content of 14 antioxidants (gallic acid, caffeic acid, coumaric acid, coutaric acid, ferulic acid, fertaric acid, trans-piceid, trans-piceatannol, rutin, quercetin-3-ß-d-glucoside, quercitrin, myricetin, catechin, and epicatechin) were studied. The results of the study show the high content of antioxidant components in grape seeds and the differences in content in individual varieties and in individual years.

Solid phase adsorption of emodin on hydrotalcites and inorganic oxides: A preliminary study.

Solid phase extraction applied to plant matrices is nowadays a well validated technique allowing the concentration and purification of selected secondary metabolites for subsequent analysis. In this short communication we screened the efficiency of 16 selected solid supports including layered structures (hydrotalcites and zirconium phosphate), magnesium oxide and hydroxide, and finally the phyllosilicates talc and bentonite for the selective concentration of the anthraquinone emodin from raw solid extracts of Polygonum cuspidatum Siebold & Zucc. (sin. Reynoutria japonica Houtt.) (Polygonaceae), commonly known as "Japanese knotweed". An ethanolic solution of sample extract from this plant was vigorously mixed with fixed quantities of each solid support. Subsequent HPLC analysis, coupled to photodiode array detection, revealed that, among the solid supports assayed, the hydrotalcite zinc aluminum oleate and magnesium oxide were largely the most effective to this concern. Both were able to extract emodin from the raw extract in percentages of 81.5 % and 92.4 %, respectively. The application of the title supports for the extraction and concentration in the solid phase of anthraquinones from raw plant extracts have been reported herein for the first time.

Vaccinium macrocarpon (Cranberry)-Based Dietary Supplements: Variation in Mass Uniformity, Proanthocyanidin Dosage and Anthocyanin Profile Demonstrates Quality Control Standard Needed.

Vaccinium macrocarpon (syn. American Cranberry) is employed in dietary supplements (DS) with the aim to improve urinary tract well-being. This property is linked to the antiadhesion-activity of proanthocyanidins (PACs) against uropathogenic-bacteria. However, the current European legislation has been criticized for being weak and ineffective. Indeed, recent scientific works report mislabeled, contaminated, and adulterated supplements containing dangerous or unknown compounds, or sold at toxic doses. In this work, we analysed 24 DS that claim to contain cranberry, and to have a specific dosage of PACs. Our tests included the control of the good manufacturing practice according to the European Pharmacopoeia, and the verification of the claimed dosage of PACs. Moreover, in order to confirm the real presence of cranberry in DS, chemical fingerprinting via HPLC-UV/Vis-MS/MS was employed. Our results showed that 17 DS did not comply with the uniformity test of dosage forms, and only five contained cranberry. Finally, 16 DS claimed an incorrect amount of PACs. These data suggest that several cranberry-based DS are present in the European market with insufficient quality controls. Considering that often DS are self-prescribed by consumer relying on their claim, the data obtained in this work should encourage more controls and stricter rules.

Bioactivity evaluation and HPLC UV-VIS based quantification of antioxidant secondary metabolites from extract and fractions of Bistorta amplexicaulis rhizome.

Bistorta amplexicaulis is a popular medicinal plant and reported as rich source of antioxidant compounds. The present study was designed for antioxidant and anticancer potential of polarity based fractions of B. amplexicaulis and its correlation to the secondary metabolites quantified by HPLC-UV/VIS.Crude extract was prepared by maceration method and polarity based fractions were prepared by solvent-solvent extraction. Antioxidant and anticancer potential was investigated by using various physiological and non-physiological assays while secondary metabolites rutin, naringin and quercetin present in extract and fractions were quantified by using HPLC- UV/VIS. All extracts showed Antioxidant potential but highest activity was obtained with ethyl acetate fraction (DPPH IC50 5.76±0.03 µg/ml, ABTS IC50 0.74±0.1 µg/ml, Total Antioxidant Assay 72.55±0.098 Ascorbic acid equivalents, Super oxide radical scavenging assay IC506.86±0.1909 µg/ml, Hydroxyl radical scavenging assay IC50 0.96±0.1690 µg/ml). The cytotoxicity of fractions against HepG2 cell lines showed lowest ell viability in n-hexane fraction (11%). The results revealed that ethyl acetate fraction of B. amplexicaulis can be a potential source of novel antioxidant compounds while n hexane fraction could provide anticancer compounds. A new method of simultaneous quantification of three flavonoids by using UV/VIS detector is reported in this study.

Determination of chlorpropham (CIPC) residues, in the concrete flooring of potato stores, using quantitative (HPLC UV/VIS) and qualitative (GCMS) methods.

Isopropyl-N-(3-chlorophenyl) carbamate (CIPC, common name Chlorpropham) is commonly used for post-harvest sprout inhibition in stored potatoes. It is applied as a thermal fog which results in loss to the fabric of the store and the atmosphere. Recently, there have been concerns in the United Kingdom because of cross contamination of other crop commodities that were stored in buildings with a history of CIPC usage. This cross contamination may have occurred because of retained residues in the fabric of the stores. The retention of CIPC in concrete is poorly understood; therefore the requirement for a robust analytical method for the detection and quantification of CIPC in concrete is a critical first step in tackling this problem. A method using High-Performance Liquid Chromatography with ultraviolet detection (HPLC UV/VIS) was validated. CIPC recoveries at three concentration levels (0.4, 4.0 and 40.0 µg g-1) were in the range of 90.7-97.0% with relative standard deviations between 2.14 and 3.01%. The limits of detection and quantification were 0.03 and 0.1 µg g-1, respectively. This study confirmed that CIPC was persistent in concrete to a depth of 4 cm, with >90% within the top 1 cm of the flooring.

Quantification of biologically active O-prenylated and unprenylated phenylpropanoids in dill (Anethum graveolens), anise (Pimpinella anisum), and wild celery (Angelica archangelica).

An analytical strategy based on different extraction methodologies and HPLC with spectrophotometric (UV-vis) detection has been developed to investigate the presence of and to quantitate biologically active selected unprenylated and O-prenylated phenylpropanoids, namely umbelliferone, 4'-geranyloxyferulic acid, 7-isopentenyloxycoumarin, auraptene, and umbelliprenin in dill (Anethum graveolens L.), anise (Pimpinella anisum L.), and wild celery (Angelica archangelica L.). Absolute ethanol or 7:3 water/ethanol mixtures were seen to be the most powerful extraction solvents to perform "classic" maceration or ultrasound-assisted one in terms of yields in secondary metabolites. For anethum and anise, umbelliprenine was found to be the most abundant prenyloxy secondary metabolite, while in wild celery 4'-geranyloxyferulic acid recorded the highest concentration. Our experimental approach demonstrated to be efficient for the simultaneous identification and quantitation of the above mentioned prenyloxyphenylpropanoids in the title plant species, that is reported herein for the first time in the literature.

Antioxidant Capacity of Selected Plant Extracts and Their Essential Oils.

The main objective of this study was the screening of some selected aromatic plants very popular in Greece, with respect to their total phenolic content, antioxidant capacity, reducing activity, and oxidative stability. All plants were extracted with the conventional method, reflux with methanol. The essential oils of the plants were also analyzed for their antioxidant properties. The total phenolic content was determined by the Folin-Ciocalteu method using gallic acid as the standard, while the phenolic substances were identified and quantified by High Performance Liquid Chromatography (HPLC) coupled with a multi-wavelength ultraviolet-visible (UV-vis) detector. The antioxidant capacity of the plant extracts was measured by their ability to scavenge free radicals such as (a) DPPH (2,2-diphenyl-1-picrylhydrazyl) and, (b) ABTS (2,2'-azinobis-(3-ethylbenzothiaziline-6- sulfonate). The Folin-Ciocalteu method proved the existence of antioxidants in the aromatic plant extracts. Taking into account the results of the DPPH and ABTS methods, the free radical scavenging capacity was confirmed. Eventually, all plants exhibited low but noticeable protection levels against lipid oxidation, as determined by the Rancimat test.