High-performance Liquid Chromatography Chromatograms Research Articles

Antioxidant and Anti-Inflammatory Activities of Ethanol Extract and its Fractions from Acer tegmentosum Stem

In this study, the first purpose was to determine total phenolic, total flavonoid and total terpenoid content of the Acer tegmentosum ethanol extract and four different types of fractions, including chloroform, ethyl acetate, n-butanol and water. Among the samples, Acer tegmentosum ethanol extract and fraction of ethyl acetate showed the highest total phenolic, total flavonoid and total terpenoid content. The second purpose was to evaluate the antioxidant activity of the samples assessed by in vitro methods such as scavenging capacity of 2,2-diphenyl-1-picryl-hydrazyl-hydrate and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid free radicals. Especially, Acer tegmentosum ethanol extract and fraction of ethyl acetate showed the strongest scavenging capacity with the lowest half maximal scavenging capacity values of these free radicals. The third purpose was to evaluate the anti-inflammatory activity in the lipopolysaccharide-stimulated Ralph and William’s cell line 264.7 macrophages treated with various concentrations of Acer tegmentosum ethanol extract and fraction of ethyl acetate, and no cytotoxic effect on the macrophages was observed at the concentration of 100 μg/ml of this extract. It suggested that this extract suppresses the activation of macrophages to secrete both pro-inflammatory cytokines (interleukin-1 beta, tumour necrosis factor alpha, inducible nitric oxide synthase and interleukin-6) and enzyme (inducible nitric oxide synthase) and anti-inflammatory cytokine (interleukin-10). The fourth purpose was to identify some compounds in a sample responsible for the antioxidant and anti-inflammation activities. In high-performance liquid chromatography chromatograms, the peak retention times detected from Acer tegmentosum ethanol extract and fraction of ethyl acetate in comparison with those from reference standards identified six compounds such as gallic acid, salidroside, (-)-epigallocatechin, (+)-catechin, scopoletin and trans-ferulic acid. It suggested that these compounds are responsible for antioxidant and anti-inflammation activities. Taken together, Acer tegmentosum seems to be one of the most promising species as a therapeutic source against diseases.

α-glucosidase inhibitors from Syzygium polyanthum (Wight) Walp leaves as revealed by metabolomics and in silico approaches

Ethnopharmacological relevanceSyzygium polyanthum (Wight) Walp leaves are traditionally used to cure diabetes in many regions of Indonesia. Traditional use involves boiling the leaves until the water is reduced to half volume, and then the decoction is taken 1–2 times daily. Despite several studies reporting the antidiabetic activity of this plant, bioactive compounds have not been well identified. Aim of the studyIndonesia is one of the countries with the highest diabetes cases, particularly type 2 diabetes mellitus (T2DM). Few people have access to modern medicinal treatment; thus, the role of antidiabetic traditional medicine has become increasingly important. This research aimed to identify α-glucosidase inhibitors from S. polyathum leaves using a metabolomics approach. When the active compounds of S. polyathum are properly identified, the quality of the herb can be more easily controlled. Materials and methodsThe dried leaves of S. polyanthum were extracted by a comprehensive extraction method using a solvent combination of n-hexane, acetone, and water in a gradient, resulting in a total of 42 fractions. All fractions were subjected to an in vitro α-glucosidase inhibition test and chemical profile analysis using Nuclear Magnetic Resonance (NMR) and high performance liquid chromatography (HPLC). Orthogonal projection least square (OPLS) analysis was used to correlate the two data to identify NMR signals, and HPLC chromatogram peaks correlated to the activity. 2D NMR and ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) analyses were also used to give more precise compound identification. The activity of the identified active compounds was confirmed by an in silico technique. Results and discussionThe results of the α-glucosidase activity test showed that the most active fractions were obtained from solvents with medium polarity: Fractions 9 and 10 (F9 and F10), obtained from gradient acetone-water 4:1 and 3:2, respectively. The IC50 values of F9 and F10 were 24.8 and 31.8 μg/mL, respectively. NMR data showed that F9 had more intense and diverse signals in the aromatic region than F10. OPLS analysis results showed that some typical flavonoid signals abundant in F9 positively correlated with α-glucosidase activity. 2D NMR and UHPLC-HRMS analysis of F9 led to the conclusion that these signals could be attributed to myricetin-3-O-rhamnoside (myricitrin) and epigallocatechin-3-gallate (EGCG). In silico analysis confirmed these results, as myricitrin and EGCG had binding energies resembling acarbose as a positive control (-8.47, -8.19, and -10.13, respectively). ConclusionsNMR and HPLC-metabolomics successfully identified myricitrin and EGCG as α-glucosidase inhibitors from S. polyanthum leaves, and docking analysis validated their inhibitory activity. The results of this study justified the traditional use of S. polyanthum as an antidiabetes herbal.

Nickel-Catalyzed Reductive Asymmetric Aryl-Acylation and Aryl-Carbamoylation of Unactivated Alkenes

Nickel-Catalyzed Reductive Asymmetric Aryl-Acylation and Aryl-Carbamoylation of Unactivated Alkenes

Jianpiyiqi formula ameliorates chronic atrophic gastritis in rats by modulating the Wnt/β-catenin signaling pathway.

Jianpiyiqi formula is a Traditional Chinese Medicine (TCM) prescription and is used for the clinical treatment of patients with chronic atrophic gastritis (CAG). The aim of the present study was to examine the underlying mechanisms of Jianpiyiqi formula treatment for CAG via the Wnt/β-catenin signaling pathway. The high-performance liquid chromatography (HPLC) chromatogram of Jianpiyiqi formula was constructed. A CAG rat model induced by N-methyl-N'-nitro-N-nitrosoguanidine and ranitidine was established. The body weight and food intake of the rats was recorded and rat gastric morphology was visually examined. Pathological analysis of rat gastric tissue was also performed. The levels of gastrin (GAS), pepsin (PP), somatostatin (SS) and prostaglandin E2 (PGE2) in rat serum were detected using ELISAs. The expression levels of proteins and genes associated with the Wnt/β-catenin signaling pathway were measured via immunohistochemistry and reverse transcription-quantitative PCR. The HPLC chromatogram of Jianpiyiqi formula was determined and as active components, liquiritin and hesperidin were identified from the chromatogram. Compared with the blank group, the body weight and feed intake of the rats were decreased, and gastric mucosal atrophy and inflammation appeared in the model group. Treatment with Jianpiyiqi formula increased the body weight and feed intake of the rats, as well as relieved the gastric atrophy and inflammation. The contents of GAS, PP, SS and PGE2 were significantly reduced in the model group compared with the blank group. Jianpiyiqi formula significantly increased GAS, PP, SS and PGE2 levels in serum of rats with CAG. In the model group, Wnt1, β-catenin and cyclin D1 protein expression levels were increased, and glycogen synthase kinase-3β (GSK-3β) protein expression levels were decreased. Jianpiyiqi formula decreased the protein expression levels of Wnt1, β-catenin and cyclin D1 and increased the protein expression levels of GSK-3β. Compared with the blank group, the mRNA expression levels of Wnt1, Wnt5a, β-catenin, cyclin D1 and MMP7 were upregulated, and the mRNA expression levels of GSK-3β were downregulated in the model group. Treatment with Jianpiyiqi formula downregulated the mRNA expression levels of Wnt1, Wnt5a, β-catenin, cyclin D1 and MMP7 and upregulated the mRNA expression levels of GSK-3β. All of the experimental results indicated that Jianpiyiqi formula exerted a therapeutic effect on rats with CAG and inhibited the activation of the Wnt/β-catenin signaling pathway. Thus, Jianpiyiqi formula, as an effective TCM prescription for treating patients with CAG, may be more widely used in the clinic.

Assessment of green biodiesel production based on Egyptian waste cooking oil using different porous membranes

In this study, green biodiesel was produced from Egyptian waste cooking oil by trans-esterification reaction using membrane technology. The produced biodiesel was characterized by physicochemical properties, FTIR, and gas chromatography/mass spectrometry (GC/MS). It was found that it meets the specifications of the American standard test method (ASTM) D6751. Because glycerol was the only impurity that was difficult to separate from the prepared biodiesel by regular methods, polysulfone (PS) membrane and three hybrid membranes were prepared using the phase inversion method for the glycerol purification process. The hybrid membranes were prepared by mixing PS with different weight ratios of polyethylene glycol (PEG), polyvinyl pyrrolidone (PVP), and zinc oxide nanoparticles (ZnO NPs). The structure and surface morphology of the resulted membranes were characterized by scanning electron microscope (SEM), Fourier-transform infrared spectroscopy (FTIR), and thermal gravimetric analysis (TGA). The purification efficiency of these membranes was satisfied by high-performance liquid chromatography (HPLC) and weight loss ratio. The HPLC chromatogram showed that the membrane of the PS/ZnO NPs provided the highest separation rate of the biodiesel by-product glycerol compared to the other ones.

ATR-FTIR Spectroscopy, HPLC Chromatography, and Multivariate Analysis for Controlling Bee Pollen Quality in Some Algerian Regions.

Bee pollen collected by honeybees (Apis mellifera) is one of the bee products, and it is as valuable as honey, propolis, royal jelly, or beebread. Its quality varies according to its geographic location or plant sources. This study aimed to apply rapid, simple, and accurate analytical methods such as attenuated total reflectance Fourier transform infrared spectroscopy (ATR–FTIR) and high-performance liquid chromatography (HPLC) along with chemometrics analysis to construct a model aimed at discriminating between different pollen samples. In total, 33 samples were collected and analyzed using principal component analysis (PCA), hierarchical clustering analysis (HCA), and partial least squares regression (PLS) to assess the differences and similarities between them. The PCA score plot based on both HPLC and ATR–FTIR revealed the same discriminatory pattern, and the samples were divided into four major classes depending on their total content of polyphenols. The results revealed that spectral data obtained from ATR–FTIR acquired in the region (4000–500 cm–1) were further subjected to a standard normal variable (SNV) method that removes scattering effects from spectra. However, PCA, HCA, and PLS showed that the best PLS model was obtained with a regression coefficient (R2) of 0.9001, root-mean-square estimation error (RMSEE) of 0.0304, and root-mean-squared error cross-validation (RMSEcv) of 0.036. Discrimination between the three species has also been possible by combining the pre-processed ATR–FTIR spectra with PCA and PLS. Additionally, the HPLC chromatograms after pre-treatment (SNV) were subjected to unsupervised analysis (PCA–HCA) and supervised analysis (PLS). The PLS model confers good results by factors (R2 = 0.98, RMSEE = 8.22, and RMSEcv = 27.86). Prospects for devising bee pollen quality assessment methods include utilizing ATR–FTIR and HPLC in combination with multivariate methods for rapid authentication of the geographic location or plant sources of bee pollen.

Laccase treatment of phenolic compounds for bioethanol production and the impact of these compounds on yeast physiology

Laccase treatment of phenolic compounds found in lignocellulosic hydrolysates is an alternative to reduce the growth inhibitory effect of these compounds on fermenting microorganisms. In order to determine the main factors that affect the efficiency of this biocatalytic approach, laccase oxidation of individual and mixtures of phenolic compounds was evaluated. Additionally, the effect of phenolic compounds on Saccharomyces cerevisiae cell physiology and their effect on yeast membrane liposome models were evaluated. Trametes versicolor laccase showed complete oxidation of six phenolic compounds, while Agaricus bisporus laccase showed oxidation percentages ranging between 30–100%. High Performance Liquid Chromatography (HPLC) analyses suggested that compounds such as syringic acid, catechol and gallic acid were not completely polymerised after laccase treatment, which could explain the increase of negative effects observed when they were added to S. cerevisiae cultures. Ferulic acid and vanillin oxidation led to a 6.4 and 6.5-fold increase in ethanol production, respectively, compared with the untreated cultures. In phenolic mixtures, chemical interactions between phenolic compounds led to biotransformation of these compounds since different by-products were observed in HPLC chromatograms. Moreover, after laccase treatment, the negative effects of these mixtures were reduced, resulting in an increase in ethanol production yields. No correlation was observed between growth inhibition data, the effect on liposomes and the detoxification efficiencies. Thus, the efficiency of laccase detoxification was not related to the toxicity of the compounds but could be conditioned by the heterogeneity in the chemical structure and properties of these compounds (functional side groups, hydrophobicity, pKa).

High-performance liquid chromatography (HPLC) analysis for flavonoids profiling of Napier grass herbal tea

Natural plant products are becoming more and more essential in helping to promote safe well-being worldwide. This leads to a substantial rise in the consumption of various herbal teas. The presence of beneficial bioactive ingredients such as flavonoids may be correlated with Napier grass herbal tea having health benefits associated with their intake. Studies have shown that herbal teas have preferentially high antioxidant activity due to the presence of flavonoids in them. The purpose of this study was to identify the Napier grass herbal teas flavonoids prepared under different conditions. Napier grass herbal teas have been formulated using green tea and black tea processes, respectively. The tea samples were extracted in water (95°C, 30 mins) and 60% (v/v) aqueous methanol (30 mins), respectively. Approximately, 1% (v/v) aqueous acetic acid solution (solvent A) and acetonitrile (solvent B) were used as the mobile phase. The flow rate was adjusted to 0.7 mL/min, the column was thermostatically controlled at 28°C, and the injection column was kept at 20 μL. HPLC chromatograms were detected using a photodiode array UV detector at 272 nm. Gallic acid, P-coumaric acid, catechin, epigallocatechin gallate, quercetin, rutin, myricetin and kaempferol were found in both Napier grass water and methanolic extracts, respectively. The findings suggested that the HPLC techniques are ideal for the detection and identification of flavonoids in Napier grass teas

Impact on degradation of antibiotics from poultry litter using Autothermal Thermophilic Aerobic Digestion (ATAD).

Tetracycline (TC) is one of the common antibiotics which is widely used in livestock growth promotion. The prevalent application TC may pave way to progression of antibiotic resistant bacteria. The main objective of this study is to determine the effect of Autothermal Thermophilic Aerobic Digestion (ATAD) on the fate of TC residues found in digested poultry litter. For the determination of TC in poultry litter, thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) were done. TLC result revealed that the Rf value of standard TC on TLC plate was 0.97 which correlates with the Rf value of TC at 0, 12, 24 and 36 h of digested poultry litter sample and not at 48, 60 and 72 h. HPLC chromatogram revealed that the limits of detection and the recovery were 5 µg/kg and 96% for standard TC. Linear correlation curves were obtained over the series of 100–500 µg/mL with correlation coefficient of 0.996 and the calibration curve was Y = 0.001X + 0.066. These results confirmed the degradation of TC in ATAD digestion of poultry litter by abiotic processes.

Evaluation of the Long-Term Storage Stability of the Cyanide Antidote: Dimethyl Trisulfide and Degradation Product Identification.

This study reports the long-term storage stability of a formulation of the cyanide (CN) antidote dimethyl trisulfide (DMTS). The F3-formulated DMTS was stored in glass ampules at 4, 22, and 37 °C. Over a period of one year, nine ampules (n = 3 at each temperature) were analyzed by high-performance liquid chromatography (HPLC)–UV/vis at daily time intervals in the first week, weekly time intervals in the first month, and monthly thereafter for a period of one year to determine the DMTS content. No measurable loss of DMTS was found at 4 and 22 °C, and good stability was noted up to five months for samples stored at 37 °C. At 37 °C, a 10% (M/M) decrease of DMTS was discovered at the sixth month and only 30% (M/M) of DMTS remained by the end of the study; discoloration of the formulation and the growth of new peaks in the HPLC chromatogram were also observed. To identify the unknown peaks at 37 °C, controlled oxidation studies were performed on DMTS using two strong oxidizing agents: meta-chloroperoxybenzoic acid (mCPBA) and hydrogen peroxide (H2O2). Dimethyl tetrasulfide and dimethyl pentasulfide were observed as products using both of the oxidizing agents. Dimethyl disulfide was also observed as a product of degradation, which was further oxidized to S-methyl methanethiosulfonate only when mCPBA was used. HPLC–UV/vis and gas chromatography–mass spectrometry/solid phase microextraction analysis revealed good agreement between the degradation products of the stability study at 37 °C and those of disproportionation reactions. Furthermore, at 4 and 22 °C, chromatograms were remarkably stable over the one-year study period, indicating that the F3-formulated DMTS shows excellent long-term storage stability at T ≤ 22 °C.

HPLC-Based Chemometric Analysis for Coffee Adulteration.

Coffee is one of the top ten most adulterated foods. Coffee adulterations are mainly performed by mixing other low-value materials into coffee beans after roasting and grinding, such as spent coffee grounds, maize, soybeans and other grain products. The detection of adulterated coffee by high performance liquid chromatography (HPLC) is recognized as a targeted analytical method, which carbohydrates and other phenolic compounds are usually used as markers. However, the accurate qualitation and quantitation of HPLC analyses are time consuming. This study developed a chemometric analysis or called non-targeted analysis for coffee adulteration. The HPLC chromatograms were obtained by direct injection of liquid coffee into HPLC without sample preparation and the identification of target analytes. The distinction between coffee and adulterated coffee was achieved by statistical method. The HPLC-based chemometric provided more characteristic information (separated compounds) compared to photospectroscopy chemometric which only provide information of functional groups. In this study, green Arabica coffee beans, soybeans and green mung beans were roasted in industrial coffee bean roaster and then ground. Spent coffee ground was dried. Coffee and adulterants were mixed at different ratio before conducting HPLC analysis. Principal component analysis (PCA) toward HPLC data (retention time and peak intensity) was able to separate coffee from adulterated coffee. The detection limit of this method was 5%. Two models were built based on PCA data as well. The first model was used to differentiate coffee sample from adulterated coffee. The second model was designed to identify the specific adulterants mixed in the adulterated coffee. Various parameters such as sensitivity (SE), specificity (SP), reliability rate (RLR), positive likelihood (+LR) and negative likelihood (−LR) were applied to evaluate the performances of the designed models. The results showed that PCA-based models were able to discriminate pure coffee from adulterated sample (coffee beans adulterated with 5%–60% of soybeans, green mung beans or spent coffee grounds). The SE, SP, RLR, +LR and −LR for the first model were 0.875, 0.938, 0.813, 14.1 and 0.133, respectively. In the second model, it can correctly distinguish the adulterated coffee from the pure coffee. However, it had only about a 30% chance to correctly determine the specific adulterant out of three designed adulterants mixed into coffee. The SE, RLR and −LR were 0.333, 0.333 and 0.667, respectively, for the second model. Therefore, HPLC-based chemometric analysis was able to detect coffee adulteration. It was very reliable on the discrimination of coffee from adulterated coffee. However, it may need more work to tell discern which kind adulterant in the adulterated coffee.

Detection of Antioxidant Phytochemicals Isolated from Camellia japonica Seeds Using HPLC and EPR Imaging.

In this study, we investigated the formation of stable radicals and compounds related to antioxidants in Camellia japonica seeds using high performance liquid chromatography (HPLC) and X-band electron paramagnetic resonance imaging (EPRI). The C. japonica seed coat extracts exhibited antioxidant activity in both in vitro and cell-based studies. The extracts inhibited reactive oxygen and reactive nitrogen species production in cell-based studies. HPLC chromatograms indicated that hydrophilic antioxidant compounds—namely, gallic acid, gallocatechin, epigallocatechin, caffeine, catechin, epicatechin, and epicatechin gallate—were found in the methanolic extract. Lipophilic antioxidant compounds—including α-tocopherol, γ-tocopherol, β-tocopherol, δ-tocopherol, α-tocotrienol, γ-tocotrienol, and δ-tocotrienol—were found in the hexane extract. EPRI primarily detected paramagnetic species in seed coats. These radical species were stable organic radicals based on the peak-to-peak line width and g-values. The signals from these stable radicals were strong and stable with a g-value of 2.002. Noninvasive EPRI of the radicals present in C. japonica seeds indicated that the stable radicals were essentially located in the seed coats. The EPRI of the cotyledon demonstrated that additional radicals were localized at an apex of the cotyledon. The results indicated that the stable radicals detected by EPRI and the hydrophilic and lipophilic antioxidant compounds analyzed by HPLC were related to antioxidant reactants and products.

African mustard (Brassica tournefortii) as source of nutrients and nutraceuticals properties.

Brassica tournefortii is an annual herbaceous plant, native to the North Africa and Middle East. It is considered as an excellent medicinal plant due to its richness by antioxidant like isothiocyanates and polyphenols. The present study is the first phytochemical investigation on Brassica tournefortii organs (leaves, stems, and roots) in terms of nutraceutical, chemical composition, and bioactivity. Brassica tournefortii leaves exhibited the highest values of nutraceutical contents. Interestingly, gas chromatograph-y-mass spectrometry (GC-MS) analysis enabled to identify three new isothiocyanates: iberverin nitrile and iberin detected only in roots, and iberin nitrile detected in all organs. HPLC chromatograms displayed different profiles depending on organic solvent and extracted organ. Icariin and 5,7-dihydroxy 4-propylcoumarin showed the highest concentrations with 2.3 and 1.3mg/g of dr among other molecules identified by high performance liquid chromatography (HPLC). Some phenolic compounds were identified in more than one organ extracts such as phenoxodiol and 4-hydroxy-3-propylbenzoic acid methyl ester. Brassica tournefortii extracts showed a moderate total phenolic contents and anti-15-LOX activity, while they exhibited a good anti-α-glucosidase activity ranging from 40% to 60%. Furthermore, leaves-MeOH and root-dichloromethane (DCM) extracts induced the highest cytotoxicity against MCF-7 cell lines, while roots-cyclohexane (CYHA) extract highlighted the highest inhibition activity against, both, HCT-116 and OVCAR cell lines.

Authentication of the Bilberry Extracts by an HPLC Fingerprint Method Combining Reference Standard Extracts.

A simple and fast high-performance liquid chromatography (HPLC) fingerprint method combining reference standard extract for the identification of bilberry extract was developed and validated. Six batches of bilberry extract collected from different manufactures were used to establish the HPLC fingerprint. Other berry extracts—such as blueberry extracts, mulberry extracts, cranberry extracts, and black rice extracts—were also analyzed for their HPLC chromatograms. The fingerprints of five batches of bilberry extract showed high similarities, while one batch was distinguished from others. Additionally, the content of anthocyanin Cyanidin-3-O-glucoside (Cy-3-glc) in each berry extract was analyzed and compared. The results indicate that this HPLC fingerprint method, combining reference standard extracts, could be used for the authentication and quality control of bilberry extracts.

The effect of drying treatment to metabolite profile and cytotoxic potential of Rhizophora apiculata leaves

Abstract. Sibero MT, Siswanto AP, Pribadi R, Sabdono A, Radjasa OK, Trianto A, Frederick EH, Wijaya AP, Haryanti D, Triningsih DW, Hayuningrat SJ, Igarashi Y. 2020. The effect of drying treatment to metabolite profile and cytotoxic potential of Rhizophora apiculata leaves. Biodiversitas 21: 2180-2187. Coastal communities in Indonesia have utilized Rhizophora spp. leaves as a traditional medicine for many years. The previous studies have succeeded in extracting bioactive compounds from this mangrove after drying treatment, but there is a possibility of the compound decomposition or breakdown. This study aimed to determine the effect of drying treatment on the metabolite profile of R. apiculata leaves which were taken from mangrove forests in Rembang, Central Java. The effect of pre-drying treatment was examined by comparing the metabolites profiles of fresh, oven-dried, and sun-dried leaves crude extracts. Extraction was carried out using maceration method with agitation (110 r.p.m.) for 24 hours in methanol. The metabolite profile was analyzed using high-performance liquid chromatography (HPLC) with diode array detector (DAD) and thin layer chromatography (TLC), while secondary metabolites were studied by phytochemical test. The phytochemical results showed that there were no differences in metabolites in dried and fresh R. apiculata leaves. Crude extract of fresh and oven-dried gave 10 spots on the TLC, while sun-dried crude extract had 9 spots. The one missing spot (Rf value of 0.79) in the sun-dried crude extract might have unstable compounds that are easily degraded or damaged by the sunlight. Moreover, HPLC chromatogram indicated the pre-drying treatment gave alteration to the R. apiculata metabolites that only detected at 400 nm. Cytotoxic assay against P388 murine leukemia cell indicated that oven-dried treatment gave the best anticancer activity with IC50 value of 0.0323 mg/mL.

Root exudation of organic acids as affected by plant growth-promoting rhizobacteria Bacillus subtilis RR4 in rice

ABSTRACTBacillus subtilis possesses plant growth-promoting traits and enhances the level of essential metabolites in plants for improved growth. Included among the metabolites are the primary metabolite malic acid (MA) and the secondary metabolite salicylic acid (SA). The impact of plant growth-promoting rhizobacteria (PGPR) on root exudation/secretion of metabolites (like MA and SA) remains unexplored, however. In this study, we aim to analyze the impact of B. subtilis RR4 on the root exudation pattern of MA and SA in rice (Oryza sativa L.). For this, rice plants were treated with the PGPR B. subtilis RR4 in hydroponics and the root exudates were collected at different periods to analyze the changes in the levels of MA and SA through high-performance liquid chromatography (HPLC). Analysis of HPLC chromatograms showed that progressive colonization by RR4 enhanced root exudation of MA 3-fold and of SA 7-fold at 96 h post-inoculation (hpi), as compared to the control. This study, thus, highlights the potential of B. subtilis RR4 of inducing/enhancing root exudation of MA and SA, which are stress alleviators, plant growth-promoting agents, and a source of carbon for microbes. With this knowledge, bioformulations, comprising the PGPRs and specific metabolites as PGPR stimulants, can be developed.

Antihypertensive Effects of Rg3-Enriched Korean Vitamin Ginseng in Spontaneously Hypertensive Rats

Ginseng is well known to treat various diseases. Ginsenoside Rg3 exhibits a variety of pharmacological activities including cardiovascular protective effects. Vitamins utilized as supplements have minimal interactions with other drugs making them attractive targets for therapeutics. Here, we prepared Rg3-enriched Korean ginseng catalyzed by vitamin (REKVG) and evaluated its ability to improve hypertension in spontaneously hypertensive rats (SHRs). The ginsenoside content in both Korean Red ginseng (KRG) and REKVG were analyzed using high-performance liquid chromatography (HPLC). Male SHRs and Wistar-Kyoto rats (WKYs) were randomly divided into 6 groups (WKY saline, WKY KRG, WKY REKVG, SHR saline, SHR KRG, and SHR REKVG). KRG and REKVG were orally administered once daily to the rats at a dose of 10 mg/kg for 6 weeks, and blood pressure was measured in live rats using the tail-cuff method. Human umbilical vein endothelial cells were used for the in vitro experiment. HPLC chromatograms revealed that the concentration of ginsenoside Rg3 in REKVG was much higher than that in KRG. The administration of REKVG significantly decreased the systolic blood pressure in SHRs at the end of 6 weeks as compared to KRG. Further, REKVG use resulted in a dose-dependent increase in Akt and endothelial nitric oxide synthase (eNOS) phosphorylation and NO production in endothelial cells. In addition, the administration of REKVG significantly increased Akt and eNOS phosphorylation and increased plasma NO levels in SHRs. We conclude that REKVG effectively lowers the blood pressure in rats and therefore could be considered for use in preventing or improving hypertension.

Study on Chemical Modifications of Glutathione by Cold Atmospheric Pressure Plasma (Cap) Operated in Air in the Presence of Fe(II) and Fe(III) Complexes

Cold atmospheric pressure plasma is an attractive new research area in clinical trials to treat skin diseases. However, the principles of plasma modification of biomolecules in aqueous solutions remain elusive. It is intriguing how reactive oxygen and nitrogen species (RONS) produced by plasma interact on a molecular level in a biological environment. Previously, we identified the chemical effects of dielectric barrier discharges (DBD) on the glutathione (GSH) and glutathione disulphide (GSSG) molecules as the most important redox pair in organisms responsible for detoxification of intracellular reactive species. However, in the human body there are also present redox-active metals such as iron, which is the most abundant transition metal in healthy humans. In the present study, the time-dependent chemical modifications on GSH and GSSG in the presence of iron(II) and iron(III) complexes caused by a dielectric barrier discharge (DBD) under ambient conditions were investigated by IR spectroscopy, mass spectrometry and High Performance Liquid Chromatography (HPLC). HPLC chromatograms revealed one clean peak after treatment of both GSH and GSSH with the dielectric barrier discharge (DBD) plasma, which corresponded to glutathione sulfonic acid GSO3H. The ESI-MS measurements confirmed the presence of glutathione sulfonic acid. In our experiments, involving either iron(II) or iron(III) complexes, glutathione sulfonic acid GSO3H appeared as the main oxidation product. This is in sharp contrast to GSH/GSSG treatment with DBD plasma in the absence of metal ions, which gave a wild mixture of products. Also interesting, no nitrosylation of GSH/GSSG was oberved in the presence of iron complexes, which seems to indicate a preferential oxygen activation chemistry by this transition metal ion.

Ability of Three Kind of Imidazole Dipeptides, Carnosine, Anserine, and Balenine, to Interact with Unsaturated Fatty Acid-Derived Aldehydes and Carbohydrate-Derived Aldehydes

Imidazole dipeptides (IDPs) such as carnosine (CAR), anserine (ANS), and balenine (BAL) are widely distributed in the skeletal muscle of vertebrates. Recently, several studies have revealed that CAR plays an important role in the detoxification of cytotoxic aldehydes arising from the peroxide of unsaturated fatty acids and carbohydrate metabolite. Although intensive studies on the detoxification of aldehydes by CAR have been performed, few studies have focused on the effects of detoxification by ANS and BAL. To determine the potential of minor IDPs such as ANS and BAL to react with cytotoxic aldehydes, the present study was established to investigate the consumption of IDP after co-incubation with cytotoxic aldehydes using high-performance liquid chromatography (HPLC). In the case of unsaturated fatty acid-derived aldehydes such as 4-hydroxy-2-trans-nonenal (from n-6 fatty acid) and 4-hydroxy-2-trans-hexenal (from n-3 fatty acid), ANS and CAR decreased considerably after co-incubation, but BAL did not. In the case of 3-deoxyglucosone and methylglyoxal as carbohydrate metabolites, no IDPs decreased after co-incubation; however, the absorbance at 336 nm of the CAR and BAL mixtures increased dramatically in a time-dependent manner. In the case of glyceraldehyde, which is also a carbohydrate metabolite, all IDPs, especially BAL, decreased after co-incubation and a new peak, surmised to represent an IDP-glyceraldehyde adduct, appeared on the HPLC chromatogram. These results can help explain the unique function and behavior of ANS and BAL in specific species.

Accurate discrimination of Gastrodia elata from different geographical origins using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis.

Gastrodia elata from different geographical origins varies in quality and pharmacological activity. This study focused on the classification and identification of Gastrodia elata from six producing areas using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis. Before recognition analysis, a principal component analysis was applied to ascertain the discrimination possibility with high-performance liquid chromatography fingerprints. And then, boosting partial least-squares discriminant analysis and conventional partial least-squares discriminant analysis were applied in this study. Experimental results indicated that the adaptive iteratively reweighted penalized least-squares algorithm could eliminate the baseline drift of high-performance liquid chromatography chromatograms effectively. And compared with partial least-squares discriminant analysis, the total recognition rates using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis for the calibration sets and prediction sets were improved from 94 to 100% and 86 to 97%, respectively. In conclusion, high-performance liquid chromatography combined with boosting partial least-squares discriminant analysis, which has such advantages as effective, specific, accurate, non-polluting, has an edge for discrimination of traditional Chinese medicine from different geographical origins. And the proposed methodology is a useful tool to classify and identify Gastrodia elata from different geographical origins.