Bioprocessed Wheat Ingredients: Characterization, Bioaccessibility of Phenolic Compounds, and Bioactivity During in vitro Digestion

To enlarge the applications of whole wheat grain (WWG) and wheat bran (WB) as functional ingredients in foodstuffs that can promote human health, researchers have explored bioprocessing approaches to improve the bioaccessibility of phenolic compounds from these food matrices and, subsequently, their biological effects. The objective of this study was to compare the composition in nutrients, anti-nutrients, and bioactive compounds of WWG and WB, and their respective bioprocessed products: sprouted wheat (GERM) and WB hydrolysate (stabilized by spray-drying [SPD] and microencapsulated [MEC]). In addition, to evaluate the functional properties of these ingredients, the bioaccessibility of phenolic compounds and their potential antioxidant and anti-inflammatory activities were monitored in different digestion steps. GERM had increased amounts of insoluble dietary fiber, higher diversity of oligosaccharides, and higher concentration of monosaccharides, free phosphorous, and phenolic compounds than WWG. SPD had improved content of soluble dietary fiber, oligosaccharides, monosaccharides, free phosphorous, and phenolic compounds (vs. WB), whereas MEC was mainly composed of protein and had nearly 2-fold lower content of SPD components. All the ingredients showed lower amounts of phytic acid as compared with raw materials. In all samples, hydroxycinnamic acids were the most representative polyphenols followed by minor amounts of hydroxybenzoic acids and flavonoids. Gastrointestinal digestion of GERM, SPD, and MEC revealed high stability of total phenolic compounds in both gastric and intestinal phases. Hydroxycinnamic acids were the most bioaccessible compounds during digestion among the three bioprocessed wheat ingredients studied, although their bioaccessibility varied across ingredients. In this sense, the bioaccessibility of ferulic acid (FA) derivatives increased in GERM with progression of the digestion, while it was reduced in SPD and MEC up to the end of the intestinal phase. Microencapsulation of SPD with pea protein led to generally to lower bioaccessible amounts of phenolic acids. Comparison analysis of biological effects highlighted SPD for its most potent antioxidant effects in the gastrointestinal tract (3 out 4 antioxidant parameters with highest values), while no clear differences were observed with regard to in vitro anti-inflammatory activity. Overall, these results support the potential application of GERM, SPD, and MEC as functional and nutraceutical ingredients.

The study of the qualitative composition and the quantitative content of phenolic compounds in dietary supplements with lingonberry

Aim. Today, there are a lot of dietary supplements with lingonberry at the pharmaceutical market of Ukraine; therefore, the analysis and quality control of these products are relevant. In this connection, the aim of the research was to study the qualitative composition and determine the quantitative content of phenolic compounds in dietary supplements with lingonberry.Materials and methods. Such dietary supplements as “Extract of lingonberry” (MEDAGROPROM), “Lingonberry” (Danikafarm), “Lingonberry nano” (LSS SYSTEM) were chosen for the study. The qualitative analysis was performed by thin layer chromatography (TLC), spectrophotometry was used for the quantitative determination.Results and discussion. Hydroquinone derivatives, flavonoids and hydroxycinnamic acids were found in the dietary supplements analyzed. The total content of phenolic compounds was 8.70, 0.26, 0.30 %, flavonoids – 6.37, 0.15, 0.12 %, hydroxycinnamic acids – 0.94, 0.06, 0.13 %, and hydroquinone derivatives – 1.01, 0.04, 0.03 % in such dietary supplements as “Extract of lingonberry” (MEDAGROPROM), “Lingonberry” (Danikafarm), “Lingonberry nano” (LSS SYSTEM), respectively. Conclusions. The qualitative and quantitative analysis of the dietary supplements with lingonberry analyzed has been performed. “Extract of lingonberry” (MEDAGROPROM) dietary supplement meets the requirements of the State of Pharmacopoeia of Ukraine 2.0, whereas “Lingonberry” (Danikafarm) and “Lingonberry nano” (LSS SYSTEM) do not. Based on the results of the study it can be concluded that the problem of compliance of dietary supplements is relevant today and requires the introduction of regulatory documentation for the detection and determination of biologically active substances in dietary supplements.

Selection of the optimal extractant for the extraction of phenolic compounds from Laurus nobilis L. leaves

Aim. To select the optimal extractant for the extraction of a number of groups of phenolic compounds from Lаurus nоbilis L. leaves based on the determination of the quantitative content of these groups in the extracts obtained. Materials and methods. Lаurus nоbilis L. leaves were harvested in November 2020 from artificially cultivated specimens aged 5 – 7 years. The quantitative determination of polyphenols was performed using the spectrophotometric method at a wavelength of 760 nm in accordance with the requirements of the Supplement of the State Pharmacopoeia of Ukraine (SPhU) 1.2 (2.8.14). The quantitative content of this group of compounds was calculated with reference to pyrogalol and dried substance. The quantitative determination of the amount of hydroxycinnamic acids was performed according to the SPhU 2.2 monograph “Orthosyphon stamen (kidney tea) leavesN” by the spectrophotometric method at a wavelength of 505 nm and calculated with reference to rosmarinic acid. The quantitative determination of the amount of flavonoids was performed according to the SPhU 2.0 monograph “Hawthorn leaves and flowers” by the spectrophotometric method at a wavelength of 410 nm and calculated with reference to hyperoside.Results and discussion. The analysis of the results for the quantitative determination of polyphenols, the amount of hydroxycinnamic acids and the amount of flavonoids in extracts from Lаurus nоbilis L. leaves obtained using water, water-ethanol mixtures and 96 % ethanol showed that 70 % ethanol was optimal for extracting compounds of these groups. The quantitative content of polyphenols (calculated with reference to pyrogalol) was not less than 21 %, the amount of hydroxycinnamic acids (calculated with reference to rosmarinic acid) – not less than 3 %, the amount of flavonoids (calculated with reference to hyperoside) – not less than 5 %. Conclusions. For the first time, the quantitative content of a number of groups of phenolic compounds (polyphenols, the total amount of hydroxycinnamic acids, and the total amount of flavonoids) in Lаurus nоbilis L. leaves extracts obtained using various extractants has been determined. On this basis, the optimal extractant – 70 % ethanol for the extraction of phenolic compounds has been selected. The results obtained will be used when developing the technology for obtaining substances from Lаurus nоbilis L. leaves.

Determination of the quantitative content of some groups of phenolic compounds in tinctures from the raw material of plant families Polygonaceae, Rosaceae, Asteraceae

Aim. To determine the quantitative content of total polyphenols and the amount of hydroxycinnamic acids in the series of tinctures from rhizomes with roots of Rumex confertus Willd., Sanguisorba officinalis L., roots of Rosa majalis Herrm., Rosa canina L., Arctium lappa L., Arctium minus (Hill) Bernh., Arctium tomentosum Mill., and the herb of Bidens tripartita L.Materials and methods. As study objects the tinctures from rhizomes with roots of Rumex confertus, Sanguisorba officinalis, roots of Rosa majalis, Rosa canina, Arctium lappa, Arctium minus, Arctium tomentosum and the herb of Bidens tripartita were used. These tinctures were obtained by the method of maceration at room temperature and the ratio of 1 : 5 of the plant raw material/finished products; the extractant was 50 % ethyl alcohol. The quantitative content of total polyphenols and the amount of hydroxycinnamic acids was determined by spectrophotometry according to the methods of the State Pharmacopoeia of Ukraine.Results and discussion. The limits of the quantitative content of total polyphenols and the amount of hydroxycinnamic acids in the tinctures were determined. They were not less than 0.070 mg mL-1 and 0.002 mg mL-1 for the tincture of rhizomes with roots of Rumex confertus, 0.100 mg mL-1 and 0.005 mg mL-1 for the tincture of rhizomes with roots of Sanguisorba officinalis, 0.070 mg mL-1 and 0.002 mg mL-1 for the tincture of Rosa majalis roots, 0.080 mg mL-1 and 0.001 mg mL-1 for the tincture of Rosa canina roots, 0.01 mg mL-1 and 0.001 mg mL-1 for the tincture of Arctium lappa roots, 0.010 mg mL-1 and 0.002 mg mL-1 for the tincture of Arctium minus roots, 0.001 mg mL-1 and 0.002 mg mL-1 for the tincture of Arctium tomentosum roots, 0.070 mg mL-1 and 0.001 mg mL-1 for the tincture of Bidens tripartita herb, respectively.Conclusions. The quantitative content of total polyphenols and the amount of hydroxycinnamic acids in tinctures from rhizomes with roots of Rumex confertus, Sanguisorba officinalis, roots of Rosa majalis, Rosa canina, Arctium lappa, Arctium minus, Arctium tomentosum and the herb of Bidens tripartita have been determined. The data obtained will be used in further work on the study of tinctures of these types of the plant raw material.

Synthesis of hydroxycinnamoyl shikimates and their role in monolignol biosynthesis

Hydroxycinnamoyl shikimates were reported in 2005 to be intermediates in monolignol biosynthesis. 3-Hydroxylation of p-coumarate, originally thought to occur via coumarate 3-hydroxylase (C3H) from p-coumaric acid or its CoA thioester, was revealed to be via the action of coumaroyl shikimate 3′-hydroxylase (C3′H) utilizing p-coumaroyl shikimate as the substrate, itself derived from p-coumaroyl-CoA via hydroxycinnamoyl-CoA: shikimate hydroxycinnamoyltransferase (HCT). The same HCT was conjectured to convert the product, caffeoyl shikimate, to caffeoyl-CoA to continue on the pathway starting with its 3-O-methylation. At least in some plants, however, a more recently discovered caffeoyl shikimate esterase (CSE) enzyme hydrolyzes caffeoyl shikimate to caffeic acid from which it must again produce its CoA thioester to continue on the monolignol biosynthetic pathway. HCT and CSE are therefore monolignol biosynthetic pathway enzymes that have provided new opportunities to misregulate lignification. To facilitate studies into the action and substrate specificity of C3H/C3′H, HCT, and CSE enzymes, as well as for metabolite authentication and for enzyme characterization, including kinetics, a source of authentic substrates and products was required. A synthetic scheme starting from commercially available shikimic acid and the four key hydroxycinnamic acids (p-coumaric, caffeic, ferulic, and sinapic acid) has been developed to provide this set of hydroxycinnamoyl shikimates for researchers.

Phenolic Compounds and Hepatoprotective Activity of Chicory Herb Extract

Introduction. Chicory (Cichorium intybus L.) is widely applied for liver disease treatment by traditional medicine of different countries; as well, it is the object for pharmacological research of hepatoprotective activity. In this regard, the method for obtaining dry extract of wild chicory herb (WCHE) is developed in the All-Russian Research Institute of Medicinal and Aromatic Plants.Aim. Aim of the research is determination of the qualitative composition of phenolic compounds, identification of the substances prevailing in WCHE and conducting pharmacological screening of the extract.Materials and methods. WCHE chemical composition has been explored with HPLC-MS/MS method; the main components were determined quantitatively with HPLC-UF method using single compounds that were isolated by us earlier and identified by NMR spectroscopy. WCHE pharmacological screening of hepatoprotective activity research was involving 50 male rats. Acute toxic hepatitis in animals was induced by a single subcutaneous injection of 50 % oily solution of tetrachloromethane (TCM) at a dosage of 0.4 ml per 100 g body weight. One hour before administration TCM, animals received WCHE at the doses of 100 or 500 mg/kg. 48 hours after TCM administration, the activity of serum enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), as well as the content of total bilirubin were determined for preliminary establishment of pharmacological activity. Pathomorphological studies of rat liver were carried out using histological methods. The liver histological structure was inspected using liver sections stained with hematoxylin and eosin.Results and discussion. The component composition of WCHE is represented by oxycoumarins, hydroxycinnamic acids and flavonoids. The dominant phenolic compounds are esculetin, chicoriin, chicoric, chlorogenic and caftaric acids. It was found under acute experimental toxic hepatitis, that preliminary WCHE administration reduces the toxic TCM effect on liver cells. In animals treated with WCHE at doses both 100 mg/kg and 500 mg/kg body weight, it was observed decreases in ALT activity by 35 % and 45 %, AST by 15 % and 28 %, alkaline phosphatase by 15 % and 21 %; the content of total bilirubin by 20 % and 29 %, respectively, in comparison with similar indicators in the group of animals that were not treated with the extract. The histological study showed that WCHE administration to animals at the doses of 100 and 500 mg/kg reduces dystrophic changes in hepatocytes, this effect is more pronounced at the extract dosage of 500 mg/kg.Conclusion. Main WCHE components are oxycoumarins (esculetin, chicoriin), hydroxycinnamic acids (chicoric, chlorogenic and caftaric). According to the results of screening studies, it was established that WCHE in doses of 100 mg/kg and 500 mg/kg is a promising object for further pharmacological research.

Hydroxycinnamic Acids and Their Related Synthetic Analogs: An Update of Pharmacological Activities

: The hydroxycinnamic acid scaffold is extremely versatile with various biological activities. This review will highlight the progress of the biological activities of hydroxycinnamic acids and their related synthetic analogs, including recently reported anti-cancer, anti-inflammatory, and antioxidant activities.

Parameters of obtaining tincture from underground organs of Sanguisorba officinalisand study its biological activity

The aim of the work is to experimentally determine the optimal parameters for obtaining a tincture of rhizomes with roots of Sanguisorba officinalis and to investigate its antimicrobial and antioxidant activity. Materials and methods. Rhizomes with roots of Sanguisorba officinalis were harvested in autumn 2019. When establishing the optimal parameters of the technology of obtaining tincture, the evaluation criterion was the number of the sum of polyphenols and the amount of hydroxycinnamic acids, which were determined by spectrophotometric method according to the methods of the State Pharmacopoeia of Ukraine 2.0. Antimicrobial activity was studied by agar diffusion and serial dilutions, antioxidant – in vitro using a stable radical 2,2-diphenyl-1-picrylhydrazyl (2,2-diphenyl-1-picrylhydrazyl – DPPH). Results. The optimal parameters for obtaining a tincture of rhizomes with roots of Sanguisorba officinalis was extraction method maceration at room temperature, raw material-finished product ratio 1:5, time 48 hours, extractant 50 % ethyl alcohol, the number of polyphenols, and the number of hydroxycinnamic acids in terms of dry raw materials not less than 4.0 % and 1.5 % respectively. The resulting tincture exhibits antimicrobial activity against microorganisms Bacillus subtilis ATCC 6633 (growth retardation 26.40 ± 1.04 mm), Staphylococcus aureus ATCC 25923 (growth retardation 24.60 ± 0.68 mm) and Pseudomonas aeruginosa ATCС 27853 (growth retardation 23.60 ± 0.68 mm) and at a dose of 0.02 ml showed antioxidant activity at the level of 70 %. Conclusions. The parameters of obtaining a tincture of rhizomes with roots of Sanguisorba officinalis were determined and its antimicrobial and antioxidant activity was studied.

Study of the effect of dermatological phytogel on the ability of microorganisms to form a biofilm

The development of many chronic infections, including skin diseases, is caused by bacteria growing in the form of biofilms. Bacterial biofilms provide beneficial survival mechanisms that determine virulence, disease pathogenesis, or resistance of the pathogen to antibiotics. As shown by a large number of studies, biofilms play an important role in the pathogenesis of dermatological diseases, including atopic dermatitis. The close relationship between the microbial biofilm that colonizes the skin surface and the negative consequences for human health makes the skin microbiome an object of therapeutic intervention in dermatological pathogenic processes. The work aims to study the effect of dermatological phytogel on the ability of microorganisms to form biofilms. The objects of research were samples of gel containing dry walnut leaf extract with the sum of tannins in terms of gallic acid and dry matter 30 mg/100 g of gel, dry nettle extract with the sum of hydroxycinnamic acids in terms of chlorogenic acid, and dry matter 20 mg/100 g of gel, dry thyme extract with the sum of flavonoids in terms of rutin and dry matter 35 mg/100 g of gel both monocomponent and combined. The study of the ability of individual plant components of phytogel samples N 1, N 2, N 3 and samples of combined phytogel N 4, N 5, and N 6 to influence biofilm formation have shown that the most pronounced decceleration of biofilms formation was registered in the gel sample with phytocomplex N 4 and was 19.7–20.7% to S. aureus, E. coli, P. aerugenosis and C. albicans respectively. The activity of the gel sample with phytocomplex N 4 was 1.3–1.4 times higher than that of monocomponent gel samples N 1, N 2, and N 3. When determining the ability of the test samples to destroy biofilms, it has been found that the gel sample with phytocomplex N 4 showed the greatest activity, which exceeded the specified properties of samples N 5 and N 6 by an average of 1.2 and 1.8 times. The activity of single-component gel samples N 1, N 2, and N 3 was lower in S. aureus, E. coli, P. aerugenosis and C. albicans biofilm destruction. The conducted studies prove the feasibility of further study of the combined gel with the phyto complex № 4 containing dry walnut leaf extract with the sum of tannins in terms of gallic acid and dry matter 30 mg/100 g of gel, dry nettle extract with the sum of hydroxycinnamic acids in terms of chlorogenic acid, and dry matter 20 mg/100 g of gel, dry thyme extract with the sum of flavonoids in terms of rutin and dry matter 35 mg/100 g of gel.

Enzyme Complexes of Ptr4CL and PtrHCT Modulate Co-enzyme A Ligation of Hydroxycinnamic Acids for Monolignol Biosynthesis in Populus trichocarpa

Co-enzyme A (CoA) ligation of hydroxycinnamic acids by 4-coumaric acid:CoA ligase (4CL) is a critical step in the biosynthesis of monolignols. Perturbation of 4CL activity significantly impacts the lignin content of diverse plant species. In Populus trichocarpa, two well-studied xylem-specific Ptr4CLs (Ptr4CL3 and Ptr4CL5) catalyze the CoA ligation of 4-coumaric acid to 4-coumaroyl-CoA and caffeic acid to caffeoyl-CoA. Subsequently, two 4-hydroxycinnamoyl-CoA:shikimic acid hydroxycinnamoyl transferases (PtrHCT1 and PtrHCT6) mediate the conversion of 4-coumaroyl-CoA to caffeoyl-CoA. Here, we show that the CoA ligation of 4-coumaric and caffeic acids is modulated by Ptr4CL/PtrHCT protein complexes. Downregulation of PtrHCTs reduced Ptr4CL activities in the stem-differentiating xylem (SDX) of transgenic P. trichocarpa. The Ptr4CL/PtrHCT interactions were then validated in vivo using biomolecular fluorescence complementation (BiFC) and protein pull-down assays in P. trichocarpa SDX extracts. Enzyme activity assays using recombinant proteins of Ptr4CL and PtrHCT showed elevated CoA ligation activity for Ptr4CL when supplemented with PtrHCT. Numerical analyses based on an evolutionary computation of the CoA ligation activity estimated the stoichiometry of the protein complex to consist of one Ptr4CL and two PtrHCTs, which was experimentally confirmed by chemical cross-linking using SDX plant protein extracts and recombinant proteins. Based on these results, we propose that Ptr4CL/PtrHCT complexes modulate the metabolic flux of CoA ligation for monolignol biosynthesis during wood formation in P. trichocarpa.