Performance Liquid Chromatography-electrospray Ionization Mass Research Articles

Lipidome characterisation and sex-specific differences in type 1 and type 2 diabetes mellitus

BackgroundIn this study, we evaluated the lipidome alterations caused by type 1 diabetes (T1D) and type 2 diabetes (T2D), by determining lipids significantly associated with diabetes overall and in both sexes, and lipids associated with the glycaemic state.MethodsAn untargeted lipidomic analysis was performed to measure the lipid profiles of 360 subjects (91 T1D, 91 T2D, 74 with prediabetes and 104 controls (CT)) without cardiovascular and/or chronic kidney disease. Ultra-high performance liquid chromatography-electrospray ionization mass spectrometry (UHPLC-ESI-MS) was conducted in two ion modes (positive and negative). We used multiple linear regression models to (1) assess the association between each lipid feature and each condition, (2) determine sex-specific differences related to diabetes, and (3) identify lipids associated with the glycaemic state by considering the prediabetes stage. The models were adjusted by sex, age, hypertension, dyslipidaemia, body mass index, glucose, smoking, systolic blood pressure, triglycerides, HDL cholesterol, LDL cholesterol, alternate Mediterranean diet score (aMED) and estimated glomerular filtration rate (eGFR); diabetes duration and glycated haemoglobin (HbA1c) were also included in the comparison between T1D and T2D.ResultsA total of 54 unique lipid subspecies from 15 unique lipid classes were annotated. Lysophosphatidylcholines (LPC) and ceramides (Cer) showed opposite effects in subjects with T1D and subjects with T2D, LPCs being mainly up-regulated in T1D and down-regulated in T2D, and Cer being up-regulated in T2D and down-regulated in T1D. Also, Phosphatidylcholines were clearly down-regulated in subjects with T1D. Regarding sex-specific differences, ceramides and phosphatidylcholines exhibited important diabetes-associated differences due to sex. Concerning the glycaemic state, we found a gradual increase of a panel of 1-deoxyceramides from normoglycemia to prediabetes to T2D.ConclusionsOur findings revealed an extensive disruption of lipid metabolism in both T1D and T2D. Additionally, we found sex-specific lipidome changes associated with diabetes, and lipids associated with the glycaemic state that can be linked to previously described molecular mechanisms in diabetes.

Distribution of Vicine, Convicine and Levodopa in Faba Bean Plant Tissues Determined by UltraHigh Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometry

Faba bean is an emerging pulse crop for the Canada Prairies and is becoming an increasingly important source of plant sourced protein. It also has a high content of levodopa (up to 0.1 % of the dry seed weight), a nutraceutical compound for treating Parkinson's Disease (PD). The presence of pyrimidine glycosides, vicine and convicine, however, restricts the use of faba due to their potential to cause favism in susceptible individuals. For the rapid separation and quantification of levodopa, vicine and convicine, a 5-min ultrahigh performance liquid chromatography-electrospray ionization mass spectrometry method (UHPLC-ESI MS) was developed. This method provided a wide linear range of quantification (4.8 nM to 60 µM for vicine, and 6.0 nM to 24 µM for convicine and levodopa), and low limit of detection (1.2 to 3.0 nM). The method was applied in the analysis of diverse faba bean samples, from dehulled mature faba bean and seed coat, to fresh plant tissues such as flowers, leaves, stems, roots and pods. The results indicated that fresh faba bean plant tissues, such as leaves and flowers, had the highest concentration of levodopa and low level of VC, which makes them an alternative source of plant-based l-DOPA with potential as a food-based treatment for Parkinson's Disease.

Bioaffinity ultrafiltration coupled with HPLC-ESI-MS/MS for screening potential α-glucosidase inhibitors from pomegranate peel.

Pomegranate peel (PoP) contains plenty of bioactive compounds and exhibits strong activity to prevent postprandial hyperglycaemia and improve diabetes mellitus. Presently, bioaffinity ultrafiltration coupled with high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS/MS) is employed to screen and identify the efficient α-glucosidase inhibitors in PoP and the detailed inhibitory mechanisms are further investigated. The results show that many substances, including ellagic acid, kaempferol, gallic acid, and resveratrol in PoP reveal strong activity to inhibit α-glucosidase and ellagic acid (EA) is screened as the most effective compound. Further research indicates that EA plays a competitive and reversible inhibition role against α-glucosidase with the value of Ki was 6.24 × 105 mol/L. EA also directly interacts with the amino acids of α-glucosidase mainly via van der Waals forces and hydrogen bonds, thereby, influencing the secondary structure and stability of α-glucosidase. Finally, the α-glucosidase inhibitory activity of EA is further confirmed to significantly reduce postprandial blood glucose in vivo.

Bioactive compounds of parsley (Petroselinum crispum), chives (Allium schoenoprasum L) and their mixture (Brazilian cheiro-verde) as promising antioxidant and anti-cholesterol oxidation agents in a food system

This study determined the bioactive composition and antioxidant potential of parsley, chives and their mixture (Brazilian cheiro-verde). Additionally, the effect of these herbs against cholesterol oxidation in grilled sardines (Sardinella brasiliensis) was also investigated. Ultra-high Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (UHPLC-ESI-MS) analyses revealed the presence of phenolic acids (caffeic, chlorogenic, and ferulic acids) and flavonoids (apigenin, kaempferol, catechin) in the herbs. Higher levels of phenolics (2.10 ± 0.02 mg GAE/g) and carotenoids (205.95 ± 0.17 µg/g) were determined in parsley extracts. Moreover, parsley also presented higher antioxidant capacity by DPPH (59.21 ± 0.07 %) and ORAC (109.94 ± 18.7 µM TE/g) than the other herbs. In vivo analyses demonstrated that the herbs’ extracts decreased the damage on Saccharomyces cerevisiae cells exposed to H2O2, except the chives extract at 10 μg/mL. Higher levels of cholesterol oxidation products (COPs) were determined after grilling. The total COPs increased from 61.8 ± 0.7 (raw fish) to 139.7 ± 10.1 µg/g (control). However, the addition of herbs effectively reduced cholesterol oxides formation, this effect was more pronounced in fish containing 4% parsley and 4% cheiro-verde. Promising results were found for cheiro-verde; however, it did not present synergic antioxidant effects.

Is selenium beneficial or detrimental to earthworm? Growth and metabolism responses of Eisenia Fetida to Na2SeO3 exposure

Se unevenly distributed in soils due to variations of geology and anthropogenic input, which results in different effects on earthworms. The effects of Se were characterized by analyzing the growth and metabolism responses of earthworms after exposure to three different concentrations of Na2SeO3. The results showed that except the possible growth promotion at 5 mg/kg, low and middle-level exposure to Na2SeO3 (0.3–10 mg/kg) did not significantly affect the growth of earthworms. While a significant inhibition effect on growth was observed in the high-level exposure group (30–70 mg/kg). There was an inflection point for Se performing promotion to inhibition effects on earthworm growth. To investigate the metabolic response of earthworms, a novel HPLC-ESI-MS (High Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry) method was used to determine sensitive biomarkers. Selenium exposure significantly altered the metabolism of seven essential amino acids, namely tyrosine, leucine, phenylalanine, valine, alanine, glycine, and lysine, and two selenoamino acids, namely selenomethionine and methylselenocysteine. The overall metabolism level of earthworms was not affected at low exposure concentrations, but was affected at medium and high exposure concentrations. The metabolic pathways that integrated the selenocompound metabolism and the tricarboxylic acid cycle from the perspective of energy supply and demand were affected by Na2SeO3 exposure. The derived reactive oxygen species at high exposure concentrations were probably the reason for the growth inhibition effect of Se on earthworms. This study provides biochemical insights into the effects of Na2SeO3 on earthworms and suggests that an Se concentration of about 2.3 mg/kg is appropriate for soil organism health.

Cytotoxic Properties, Anthocyanin and Furanocoumarin Content of Red-Pigments Obtained from Callistemon citrinus (Curtis) Skeels Flowers

Background and objectives: There is growing interest in introducing safe and bioactive natural red pigments to the pharmaceutical and cosmetic industries. This study was designed to determine the phytochemical content and potential cytotoxicity of red pigment from Callistemon citrinus (Curtis) Skeels (syn. Melaleuca citrina (Curtis) Dum.Cours.) flowers. Methods: The flowers’ anthocyanin rich pigment was extracted with ethanol (70%, v/v) containing 0.5% formic acid. This extract was fractionated by a three-step process through a Sep-Pak C18 cartridge with water, ethyl acetate, and methanol, respectively. The anthocyanin and coumarin content of the red pigment was identified based on a reverse phase high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS/MS). The MTT assay was used to assess the cytotoxicity of red pigment evaluated on normal human foreskin fibroblasts as well as two malignant cell lines: human breast cancer (MCF-7) and human fibrosarcoma cells (HT1080) at 24, 48 and 72 hours. Results: The anthocyanin compounds in the red pigment fraction were cyanidin (1), cyanidin 3,5-O-diglucoside (2), cyanidin 3-O-glucoside (3), and cyanidin 3-O-glucoside-8-ethyl-catechin (4). Flowers’ pigment also contained two furanocoumarins, including 8-(but-2-en-2-yl)-8,9-dihydro-2H-furo[2,3-h] chromen-2-one (or 15-methyl angenomallin, (5)) and 9-methyl-7H-furo[3,2-g] chromen-7-one (or 8-methylpsoralen, (6)). According to the MTT assays, the highest cytotoxic effect was observed on human foreskin fibroblasts with an IC50 values of <12.5 (µg/ mL, 24 h) and 85.2 (µg/ mL, 48 h). Conclusion: It might be assumed that application of the red pigment of C. citrinus in topical formulations and cosmetics should be done with caution due to the observed cytotoxicity on dermal fibroblasts.

Secondary metabolic profiles and anticancer actions from fruit extracts of immature pomegranates.

Immature fruits from Punica granatum L. thinning are a neglected side product of pomegranate production with cumbersome disposal costs for farmers. To explore value potential of immature fruits from pomegranate ‘Wonderful’ cultivars, the compositional landscapes and antitumorigenic activities of pomegranate extracts from two different stages of maturation were assessed. Cancer cell proliferation and cytotoxicity was quantified in human lung H1299 and colon HCT116 adenocarcinomas by crystal violet staining, MTS assay and caspase-3 activity. High performance liquid chromatography—diode array detector (HPLC/DAD) and high performance liquid chromatography—electrospray ionization—mass spectrometry (HPLC/ESI-MS) analyses indicate that immature fruits are rich sources of gallotannins and ellagitannins, with the highest amounts contained in immature fruit peels. Biological investigations reveal a robust anticancer activity by those immature P. granatum fruit extracts, which reflected induction of tumor cytotoxicity and cell death mechanisms. Together, present observations suggest P. granatum byproducts from the thinning process may provide unexplored values for virtuous circular economy.

Improving sesquiterpenoids production of Sarcandra glabra callus culture

Sarcandra glabra, a traditional Chinese medicine and herbal tea, has benefits from treating inflammation and fractures to curing cancer. The anti-inflammatory properties of sesquiterpenoids (chloranthalactone A, CTA; atractylenolide II, AT-II) have made S. glabra to a great potential for research and development. Here this study developed S. glabra callus culture to circumvent particular needs of the plant growing seasons and Hardiness zone, with the goal being able to produce sesquiterpenoids in vitro. The half strength Murashige and Skoog (1/2MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, 2.0 mg/L) and 6-benzylaminopurine (6-BA, 1.0 mg/L) was effective for callus induction. The 1/2MS medium supplemented with 2,4-D (2.0 mg/L) and 6-BA (0.5 mg/L) was superior for callus proliferation. The callus was friable, yellowish and rapidly grown without tissue differentiation. Moreover, high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS) analysis indicated that the contents of CTA and AT-Ⅱ in multiple subculture S. glabra callus were 3.72 ± 0.51 mg/g and 0.50 ± 0.12 mg/g in fresh weight (FW), respectively. Using abiotic stresses and elicitors, production of CTA (by ultraviolet-B radiation for 48 h) and AT-II (by methyl jasmonate, 100 μmol/L) was enhanced by 2.3-fold. In conclusion, this work not only established the S. glabra callus culture for in vitro production of CTA and AT-II, but also provided the foundation for further development and utilization of S. glabra.

Maillard Reaction Products in Gluten-Free Bread Made from Raw and Roasted Buckwheat Flour.

The formation of Maillard reaction products (MRPs) in gluten-free bread made from roasted and raw buckwheat flour was examined. The levels of phenolic compounds such as flavonoids (catechin, naringenin, quercetin, rutin, and others) and phenolic acids (like 4-hydroxybenzoic, caffeic, dihydroxybenzoic, ferulic, gallic, syringic, vanillic, and p-coumaric) were measured using reversed-phase ultra-high performance liquid chromatography-electrospray ionization mass spectrometry (RP–UHPLC–ESI-MS). Early and advanced Maillard reaction products were analyzed using HPLC, whereas spectrofluorimetric analysis was used to determine the levels of fluorescent intermediate compounds (FIC). The total levels of phenolic compounds were higher in the case of buckwheat bread prepared from roasted buckwheat flour (156 and 140 µg/g of crumb and crust, respectively). Rutin, gallic acid, and catechin were the most abundant phenolic compounds detected in roasted buckwheat bread. The roasting process resulted in significantly lower radical scavenging capacities (ABTS) of the total phenolics and flavonoids in the buckwheat bread. Taking into consideration these Maillard reaction products, we observed a significant increase in FIC level in roasted buckwheat crumb and crust (at about 40%, and 38%, respectively). At the same time, the Nε-(carboxymethyl)lysine (CML) level did not change in roasted or raw buckwheat bread crumb, though in roasted buckwheat crust the concentration of CML increased by about 21%.

Jiaotai Pill () Alleviates Insomnia through Regulating Monoamine and Organic Cation Transporters in Rats.

To reveal the effect and mechanism of Jiaotai Pill (, JTP) on insomniac rats. The insomniac model was established by intraperitoneal injection of p-chlorophenylalanine (PCPA). In behavioral experiments, rats were divided into control, insomniac model, JTP [3.3 g/(kg•d)], and diazepam [4 mg/(kg•d)] groups. The treatment effect of JTP was evaluated by weight measurement (increasement of body weight), open field test (number of crossings) and forced swimming test (immobility time). A high performance liquid chromatography-electrochemical detection (HPLC-ECD) method was built to determine the concentration of monoamine transmitters in hypothalamus and peripheral organs from normal, model, JTP, citalopram [30 mg/(kg•d)], maprotiline [40 mg/(kg•d)] and bupropion [40 mg/(kg•d)] groups. Expressions of serotonin transporter (SERT), dopamine transporter (DAT), and norepinephrine transporter (NET) were analyzed by quantitative polymerase chain reaction (qPCR) and Western blot in normal, model and JTP groups. A high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS/MS) method was established to determine the pharmacokinetics, urine cumulative excretion of metformin in vivo, and tissue slice uptake in vitro, which were applied to assess the activity of organic cation transporters (OCTs) in hypothalamus and peripheral organs. Compared with the insomniac model group, the body weight and spontaneous locomotor were increased, and the immobility time was decreased after treatment with JTP (P<0.01). Both serotonin and dopamine contents in hypothalamus and peripheral organs were increased (P<0.01). The norepinephrine content was increased in peripheral organs and decreased in hypothalamus (P<0.05 or P<0.01). At the same time, SERT, DAT, OCT1, OCT2, and OCT3 were down-regulated in hypothalamus and peripheral organs (P<0.05). NET was down-regulated in peripheral organs and up-regulated in hypothalamus (P<0.05 or P<0.01). Moreover, the activity of OCTs in hypothalamus and peripheral organs was inhibited (P<0.05). JTP alleviates insomnia through regulation of monoaminergic system and OCTs in hypothalamus and peripheral organs.

Two CYP71AJ enzymes function as psoralen synthase and angelicin synthase in the biosynthesis of furanocoumarins in Peucedanum praeruptorum Dunn.

Psoralen synthase and angelicin synthase responsible for the formation of psoralen and angelicin in Peucedanum praeruptorum Dunn were identified and functionally characterized, respectively. Furanocoumarins were reported to possess several activities such as anticancer, anti-inflammatory and neuroprotective, and function as phytotoxin and allelochemical in plants. Furanocoumarins are the main bioactive ingredient in P. praeruptorum which is a commonly used traditional Chinese medicine. Phenylalanine ammonia lyase (PAL), 4-coumarate: CoA ligase (4CL), p-coumaroyl CoA 2'-hyfroxylase (C2'H) were cloned previously to elucidate the biosynthetic mechanism of coumarin lactone ring. However, the genes involved in complex coumarins in P. praeruptorum have not been explored. Herein, putative psoralen synthase CYP71AJ49 and angelicin synthase CYP71AJ51 were cloned from P. praeruptorum. In vivo and in vitro yeast assays were conducted to confirm their activities. Furthermore, the results of High Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometry (HPLC-ESI-MS) verified that CYP71AJ49 catalyzed the conversion of marmesin to psoralen, and CYP71AJ51 catalyzed columbianetin to angelicin. Subsequently, the expression profile showed that CYP71AJ49 and CYP71AJ51 were easily affected by environmental conditions, especially UV and temperature. The genes tissue-specific expression and compounds tissue-specific distribution pattern indicated the existence of substance transport in P. praeruptorum. Phylogenetic analysis was conducted with 27 CYP71AJs, CYP71AJ49 and CYP71AJ51 were classified in I-4 and I-2, respectively. These results provide further insight to understand the biosynthetic mechanism of complex coumarins.

Molecular characterization of biomass burning tracer compounds in fine particles in Nanjing, China

Abstract In this work, a sensitive method based on the High Performance Liquid Chromatography-Electrospray Ionization-Mass spectrometry/Mass spectrometry (HPLC-ESI-MS/MS) technique was established and validated, and successfully applied to quantify biomass burning tracer species (in total 20 compounds) in fine aerosol particles (PM2.5) collected during the full year of 2016 in Nanjing, China. The annual levoglucosan concentration was determined to be 328.3 ng m−3, at a similar level as it in other Chinese cities; the concentration peaked in fall, indicating influences of crop/straw burning during harvest periods. Concentrations of other species were a few orders of magnitude lower than that of levoglucosan, with salicylic acid (1.53 ng m−3), cis-pinonic acid (0.60 ng m−3) and vanillic acid (0.51 ng m−3) as the three most abundant ones. Total concentrations of tracers indicative of softwood, hardwood and grass burning were about equal. Seasonal concentrations of these tracers were generally similar, and all peaked in winter, different from that of levolglucosan. Further positive matrix factorization (PMF) identified four types of biomass (cellulose, harwood lignin, softwood lignin and grass). Celluose was dominated in fall, while softwood and hardwood were both dominated in winter, and seasonal contributions (except summer) were relatively even to grass. On an annual basis, softwood, hardwood and grass contributions were almost on par to the selected tracers, in agreement with the results based only on concentrations, but seasonally, grass was the major one in spring, summer and fall, but much less important in winter.

Determination of 5-HT3 Receptor Antagonists in Human Urine by Porous Graphitic Carbon (PGC) Solid Phase Extraction (SPE) Coupled with High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS)

The quantitative determination of drugs in urine by high performance liquid chromatography electrospray-ionization mass spectrometry (HPLC-ESI-MS) is challenging due to complex matrix interferences and the relatively low concentrations of analytes in urine. In this study, porous graphitic carbon (PGC) was investigated as solid phase extraction (SPE) sorbent before HPLC-ESI-MS determination of three 5-hydroxytryptamine (3) (5-HT3) receptor antagonists from human urine. The factors affecting extraction of the target antagonists were investigated and optimized systematically by using single-factor tests. The optimized SPE procedure was developed for the simultaneous extraction and sample pretreatment of tropisetron, ondansetron, and granisetron in urine by HPLC-ESI-MS/MS. Using the optimized conditions, the matrix effects were reduced by the PGC-based SPE clean-up procedure, with acceptable matrix effects for tropisetron, ondansetron, and granisetron from 96.53% to 98.42%, 93.19% to 94.36%, and 94.48% to 95.60%, respectively. The reported method significantly removes matrix interferences from the urine samples and exhibited satisfactory recovery and process efficiency for the determination of tropisetron, ondansetron, and granisetron. The established pretreatment method provides a promising strategy for the removal of interferences from hydrophilic basic drugs in biological fluids.

Antioxidant Capacity of Thistle (Cirsium japonicum) in Various Drying Methods and their Protection Effect on Neuronal PC12 cells and Caenorhabditis elegans

The aim of this study was, firstly, to evaluate the phenol profile of thistle (Cirsium japonicum, CJ) by High performance liquid chromatography-electrospray ionization–mass spectrometry (HPLC–ESI–MS), dried by different methods (90 °C hot-air, 70 °C hot-air, shade-, and freeze-drying). Secondly, we aimed to evaluate the relationship between phenolic compounds content and antioxidant properties. CJ contained chlorogenic acid, linarin, and pectolinarin. Total phenolic contents of CJ significantly decreased under hot-air-drying condition, especially chlorogenic acid contents in CJ have been reduced by 85% and 60% for 90 °C and 70 °C hot-air-drying, respectively. We evaluated the protective effect on adrenal pheochromocytoma (PC12) cells and Caenorhabditis elegans using shade-dried CJ, which has the largest phenolic contents and the strongest antioxidant property. CJ-treated PC 12 cells dose-dependently exhibited the protective effects against reactive oxygen species (ROS), while cell viability increases, lactate dehydrogenase release decreases, and ROS formation decreases. Furthermore, CJ has also shown protection against ROS in C. elegans. Consequently, CJ contributed to lifespan extension under ROS stress without influencing the physiological growth.

Identification of secondary metabolites in Tripterygium wilfordii hairy roots and culture optimization for enhancing wilforgine and wilforine production

Hairy roots culture is an effective biological system to solve the shortage of natural resources of Tripterygium wilfordii for the production of the important secondary metabolites. However, there is no enough information about the secondary metabolites in Tripterygium wilfordii hairy root cultures. The present study identified the secondary metabolites in T. wilfordii hairy roots by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS). Subsequently, the nitrogen source and phosphate concentration were optimized to enhance the production of the major active compounds wilforgine and wilforine. Nine compounds including triptolide, wilforgine, wilforine, and so on were identified according to their retention behavior in liquid chromatography and mass spectrometric fragmentation patterns. The Murashige and Skoog (MS) medium was modified for enhancing wilforgine and wilforine production in T. wilfordii hairy root cultures by altering the ratio of NH4+/NO3－and the concentration of phosphate. The results showed that 10:50 mM NH4+/NO3－ and 0.3125 mM phosphate were optimal for roots growth and the production of wilforgine and wilforine. After 35 days of cultivation in the modified MS medium containing 10:50 mM NH4+/NO3－ and 0.3125 mM phosphate, the maximum production of wilforgine and wilforine was up to 1509.87 μg·flask−1 and 578.56 μg·flask−1 in T. wilfordii hairy roots separately, a 42 % and 48 % increase over that in the basic MS medium. The present study laid a foundation for the large-scale production of secondary metabolites from T. wilfordii hairy roots.

Novel ACE Inhibitory Peptides Derived from Simulated Gastrointestinal Digestion in Vitro of Sesame (Sesamum indicum L.) Protein and Molecular Docking Study.

The aim of this study was to isolate and identify angiotensin I-converting enzyme (ACE) inhibitory peptides from sesame protein through simulated gastrointestinal digestion in vitro, and to explore the underlying mechanisms by molecular docking. The sesame protein was enzymatically hydrolyzed by pepsin, trypsin, and α-chymotrypsin. The degree of hydrolysis (DH) and peptide yield increased with the increase of digest time. Moreover, ACE inhibitory activity was enhanced after digestion. The sesame protein digestive solution (SPDS) was purified by ultrafiltration through different molecular weight cut-off (MWCO) membranes and SPDS-VII (< 3 kDa) had the strongest ACE inhibition. SPDS-VII was further purified by NGC Quest™ 10 Plus Chromatography System and finally 11 peptides were identified by Nano UHPLC-ESI-MS/MS (nano ultra-high performance liquid chromatography-electrospray ionization mass spectrometry/mass spectrometry) from peak 4. The peptide GHIITVAR from 11S globulin displayed the strongest ACE inhibitory activity (IC50 = 3.60 ± 0.10 μM). Furthermore, the docking analysis revealed that the ACE inhibition of GHIITVAR was mainly attributed to forming very strong hydrogen bonds with the active sites of ACE. These results identify sesame protein as a rich source of ACE inhibitory peptides and further indicate that GHIITVAR has the potential for development of new functional foods.

High performance liquid chromatography-electrospray ionization mass spectrometric (LC-ESI-MS) methodology for analysis of amino acid energy substrates in microwave-fixed microdissected brain tissue

Hypoglycemia deprives the brain of its primary energy source glucose. Reductions in whole-brain amino acid energy substrate levels suggest that these non-glucose fuels may be metabolized during glucose shortage. Recurring hypoglycemia can cause mal-adaptive impairment of glucose counter-regulation; yet, it is unclear if amplified reliance upon alternative metabolic substrates impedes detection of continuing neuro-glucopenia. This research aimed to develop high-sensitivity UHPLC-electrospray ionization mass spectrometric (LC-ESI-MS) methodology, for complementary use with high-neuroanatomical resolution microdissection tools, for measurement of glucogenic amino acid, e.g. glutamine (Gln), glutamate (Glu), and aspartate (Asp) content in the characterized glucose-sensing ventromedial hypothalamic nucleus (VMN) during acute versus chronic hypoglycemia. Results show that VMN tissue Gln, Glu, and Asp levels were significantly decreased during a single hypoglycemic episode, and that Gln and Asp measures were correspondingly normalized or further diminished during renewed hypoglycemia. Results provide proof-of-principle that LC-ESI-MS has requisite sensitivity for amino acid energy substrate quantification in distinctive brain gluco-regulatory structures under conditions of eu- versus hypoglycemia. This novel combinatory methodology will support ongoing efforts to determine how amino acid energy yield may impact VMN metabolic sensory function during persistent hypoglycemia.

Evaluating iopamidol degradation performance and potential dual-wavelength synergy by UV-LED irradiation and UV-LED/chlorine treatment

This study revealed the iopamidol degradation performance and the synergistic effect of dual-wavelength ultraviolet light-emitting diodes (UV-LEDs). It was found that the dual-wavelength irradiation (265 and 280 nm) can promote iopamidol degradation during both UV-LED alone and UV-LED/chlorine processes (p < 0.05). The effects of key experimental parameters including pH, chlorine dosage and water matrix during UV-LED/chlorine process were evaluated. Peak degradation rate was observed at pH 7.0 due to the combined effects of the quantum yield of aquatic chlorine photolysis (Φchlorine) and absorptivity of chlorine. The destruction pathways of iopamidol in the UV-LED/chlorine process were proposed by Ultra Performance Liquid Chromatography-Electrospray Ionization-mass spectrometry. Additionally, an opposite trend of iodinated trihalomethane (I-THM) formation during subsequent chlorination between Milli-Q and real waters with prolonging irradiation time was observed, while other detected disinfection by-product (DBP) formation showed the same variation in Milli-Q and real waters. The UV-LED/chlorine treatment using dual-wavelength can enhance iopamidol degradation significantly, but it also promoted I-THM formation in real water, indicating the need for evaluating DBPs before the application of UV-LED/chlorine process in drinking water treatment.

Antioxidant Capacity and UPLC-PDA ESI-MS Phenolic Profile of Stevia rebaudiana Dry Powder Extracts Obtained by Ultrasound Assisted Extraction

Stevia leaves, which are commonly used as a natural sweetener in food products, have increased in importance for antioxidant delivery due to their high content of phenolic compounds. In this study, the influence of the drying process on stevia leaves, with regards to phenolic content and antioxidant activity during drying kinetics 40 °C for 7 h, was studied. The effect of solvent concentration and extraction time using a 32 factorial design on total phenol content (TPC), and on antioxidant activity of extracts obtained from dried stevia leaves, by ultrasound assisted extraction (UAE) as alternative method was evaluated. Steviol glycosides contents were also evaluated by a conventional and UAE method. Phenols identification, quantification and purification were performed by Ultra Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (UPLC-ESI-MS), Ultra Performance Liquid Chromatography-Photodiode Array (UPLC-PDA) and advanced automated flash purification, respectively. Drying time affected the moisture content of stevia leaves. A constant weight was reached after six hours of drying, and higher antioxidant activity was observed, while the highest TPC was obtained after seven hours of drying. The highest TPC (91.57 ± 8.8 mg GAE/g dw) and antioxidant activity (603.24 ± 3.5 μmol TE/g dw) in UAE method was obtained when ethanol 50% at 5 min was used. Steviol glycosides extracted by UAE were recorded with a content of 93.18 ± 1.36 mg/g dw and 98.97 ± 1.75 mg/g dw for stevioside and rebaudioside A respectively. Six phenolic compounds including four phenolic acids and two flavonoids were identified and quantified by UPLC-PDA, and confirmed by ESI-MS reporting its fragmentation pattern. Diosmin and chlorogenic acid were the most abundant compounds with values of 2032.36 μg/mL and 434.95 μg/mL respectively. As a novelty we found that the antioxidant activity evaluated in partially purified fractions suggested that biological activity might be attributed to the synergistic effect of the six phenols present in the stevia leaves extract. In addition to its sweeting properties, stevia leaves constitute a potential source of polyphenolic compounds, with antioxidant activity that could be used as a food additive.

Nigericin and grisorixin methyl ester from the Algerian soil-living Streptomyces youssoufiensis SF10 strain: a computational study on their epimeric structures and evaluation of glioblastoma stem cells growth inhibition

The present work describes the metabolites produced by a strain identified as Streptomyces youssoufiensis, whose secondary metabolites profile has not been studied so far. The crude ethyl acetate extract was analyzed by high performance liquid chromatography-electrospray ionization mass spectrometry, leading to the detection of the ionophoric polyethers nigericin, epinigericin, abierixin and the newly isolated grisorixin methyl ester. The presence of epimeric forms of nigericin/epinigericin and grisorixin/epigrisorixin has spurred density functional theory computational calculations. This analysis was able to provide the relative stability of the most favored epimers, setting the basis for general structural considerations applicable to several other polyethers. Both nigericin sodium salt and grisorixin methyl ester showed to affect glioblastoma stem cells proliferation in a dose-dependent manner, with a higher activity for the more lipophilic grisorixin methyl ester (GI50 values of 3.85 and 3.05 μM for VIPI and COMI human glioblastoma stem cells, respectively).