Ultra-performance Liquid Chromatography-electrospray Ionization-mass Spectrometry Research Articles

Secondary Metabolomic Analysis and In Vitro Bioactivity Evaluation of Stems Provide a Comprehensive Comparison between Dendrobium chrysotoxum and Dendrobium thyrsiflorum.

The stems of Dendrobium chrysotoxum (DC) are commonly used as health-promoting foods due to their excellent biological activities. However, the stems of D. thyrsiflorum (DT) are often used to meet the scarcity of DC in production because of their highly similar morphology. However, the related metabolomic and bioactive information on the stems of DC and DT are largely deficient. Here, secondary metabolites of DC and DT stems were identified using an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry, and their health-promoting functions were evaluated using several in vitro arrays. A total of 490 metabolites were identified in two stems, and 274 were significantly different. We screened out 10 key metabolites to discriminate the two species, and 36 metabolites were determined as health-promoting constituents. In summary, DT stems with higher extract yield, higher total phenolics and flavonoids, and stronger in vitro antioxidant activities demonstrated considerable potential in food and health fields.

Exploring the composition of Syringa reticulata subsp. amurensis seed and its underlying mechanism against chronic bronchitis

• The chemical compositions of Syringa reticulata subsp. amurensis seed were first determined with UPLC-ESI-MS. • Deciphering the underlying mechanism composition of Syringa reticulata subsp. amurensis seed against chronic bronchitis. • Six ingredients and seven targets were found against chronic bronchitis. The aim of this study was to analyze the bioactive components of Syringa reticulata subsp. amurensis (SRA) seeds, and the potential mechanism of SRA seed against chronic bronchitis is elucidated using network pharmacology. In total, 235 constituents were detected from SRA seed by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS), and 28 active ingredients were identified through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database (TCMSP), which was classified into 10 different categories including flavonoids, diterpenoids, coumarins, alkaloids, isoflavones, phenylpropanoids, sesquiterpene, anthraquinones, phenols, and lignans. Among them, 6 components vestitol(V1), wogonin(W2), cheilanthifoline(C1), miltirone(M2), glabridin(G3), and myricanone (M3) were selected as potential key constituents of SRA seed against chronic bronchitis through network pharmacology analysis. Then the 7 key targets (PTGS2, TP53, TNF, VEGFA, IL1β, IL6, and JUN) were obtained based on the target prediction and protein-protein interaction (PPI). In conclusion, we use UPLC-ESI-MS analysis and network pharmacology to reveal the chronic bronchitis mechanisms of SRA seed, which provides new theoretical research ideas for the sustainable development of endangered multifunctional plants.

Lipidomic profile changes of yellow-feathered chicken meat during thermal processing based on UPLC-ESI-MS approach

In this work, ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI-MS) was applied to compare the lipidomics profile of chicken breast meat heated to various temperatures. A total of 445 lipids including phospholipids, diacylglycerols, triacylglycerols, glycoglycerolipids, and sphingolipids were detected in chicken muscle. Significant decreases in a list of characteristic unsaturated phospholipid species including phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol were identified after heating, suggesting the occurrence of lipid oxidation, while significant increases in characteristic lysophospholipid species were found with increasing heating temperature, which were mainly derived from hydrolysis of phospholipids. Moreover, ether phospholipid and neutral lipid species exhibited remarkable changes during the heating process, and monoalk(en)yl diacylglycerol was first identified in chicken meat. The findings could contribute to the improved understanding of key lipids and biochemical reactions engaged in the heating of meat products.

Effects of Fengycins and Iturins on Fusarium oxysporum f. sp. physali and Root Colonization by Bacillus velezensis Bs006 Protect Golden Berry Against Vascular Wilt.

Bacillus velezensis Bs006 has shown antagonistic activity on Fusarium oxysporum f. sp. physali and biocontrol activity against Fusarium wilt (FW) in golden berry (Physalis peruviana). We hypothesized that strain Bs006 has the ability to synthesize antimicrobial cyclic lipopeptides (CLPs) like other members of the same species. However, if so, the real effects of CLPs on F. oxysporum f. sp. physali and their potential as a biocontrol tool against Physalis-FW have not been elucidated. In this study the CLPs profile of Bs006 in liquid culture and antagonist-plant-pathogen interactions were characterized. Also, the potential effects of supernatant free of bacteria against F. oxysporum f. sp. physali and FW were explored and compared with the effects of pure CLPs. Ultraperformance liquid chromatography-electrospray ionization-mass spectrometry analysis revealed the capacity of Bs006 to synthesize homologous compounds of iturins, surfactins, and fengycins in liquid culture and on the inhibition zone against F. oxysporum f. sp. physali in dual confrontation tests. Bs006 supernatant reduced the germination and growth of F. oxysporum f. sp. physali and caused vacuolization, swelling, and lysis of F. oxysporum f. sp. physali cells in a concentration-dependent manner. Pure fengycins affected the development of F. oxysporum f. sp. physali from 11 mg/liter and iturins from 21 mg/liter. In a gnotobiotic system, Bs006 colonized the root surface of golden berry, inhibited the growth of F. oxysporum f. sp. physali, and produced CLPs. Individual application of Bs006 and supernatant protected the plants from F. oxysporum f. sp. physali infections by 37 to 53%, respectively. Meanwhile, fengycins reduced the disease progress by 39%. These results suggest further studies to select an optimum combination of Bs006 and supernatant or CLPs, which might be a good option as biofungicide against F. oxysporum f. sp. physali.

UPLC-ESI/MS analysis of disinfection by-products (perchlorate, bromate, nitrate, nitrite and sulfite) in micro-filtered drinking water obtained from spring, well and tap water (desalinated) sources

ObjectivesIn the current study, an analytical technique has been optimized for the analysis of disinfection by-products (perchlorate (ClO4−), bromate (BrO3−), nitrate (NO3−), nitrite and sulfite (SO32–) in drinking water obtained from spring, well and tap water (desalinated) sources. MethodsThe ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI/MS) conditions have been optimized for oxyhalides ClO4− (isotopes, 37ClO4− and 35ClO4−) and BrO3− (isotopes, 81BrO3− and 79BrO3−), and inorganic anions NO3−, NO2− and SO32–. Separation was achieved by BEH C18 column with methanol (75%) and water (24.99%, HCOOH 0.01%) mobile phase at flow rate 0.2 mL/min. ResultsThe analysis was carried out in <1 min with excellent coefficient of determination, (R2 > 0.995–0.999), limit of detection (0.016–0.043 µg/L), limit of quantification (0.051–0.104 µg/L) and precision (repeatability, 1.54–2.28% and reproducibility, 3.13–4.08%) in terms of RSD%. The method was successfully applied, and the amounts of 35ClO4− (5.33–22.64 µg/L), 81BrO3− (7.52–16.73 µg/L), NO3− (5.42–12.14 mg/L), NO2− (1.08–4.37 mg/L) and SO32– (6.84–32.45 mg/L) were identified in spring water, whereas well and tap water contained 35ClO4− (4.20–21.33 µg/L), 81BrO3− (8.05–15.13 µg/L), NO3− (1.27–16.11 mg/L), NO2− (0.43–10.77 mg/L) and SO32– (7.14–36.10 mg/L), and 35ClO4− (0.89–7.37 µg/L), 81BrO3− (5.15–14.68 µg/L), NO3− (1.12–6.33 mg/L), NO2− (0.67–3.95 mg/L) and SO32– (5.40–23.26 mg/L), respectively. ConclusionsThe levels of 35ClO4−, 81BrO3−, NO3−, NO2− and SO32– were found beyond the maximum contaminant levels and drinking water equivalent levels regulated by the Environmental Protection Agency, respectively. The outcomes also revealed that comparatively tap water produced lower levels of these contaminants especially in the cities with low population densities. In addition, the geographical site would be helpful to categorize the tap water samples.

Profiling of Metabolites of Bacillus spp. and Their Application in Sustainable Plant Growth Promotion and Biocontrol

Bacillus spp. are well-characterized as efficient bioinoculants for sustainable plant growth promotion and biocontrol of phytopathogens. Members of this spp. exhibit the multifaceted beneficial traits that are involved in plant nutrition and antimicrobial activities against phytopathogens. Keeping in view their diverse potential, this study targeted the detailed characterization of three root-colonizing Bacillus strains namely B. amyloliquefaciens, B. subtilis, and B. tequilensis, characterized based on 16S rRNA sequencing homology. The strains exhibited better plant growth promotion and potent broad-spectrum antifungal activities and exerted 43–86% in-vitro inhibition of growth of eight fungal pathogens. All strains produced indole acetic acid (IAA) in the range of 0.067–0.147 μM and were positive for the production of extracellular enzymes such as cellulase, lipase, and protease. Ultra-performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (UPLC-ESI-MS/MS) analysis revealed the production of antifungal metabolites (AFMs) such as surfactins, iturins, fengycins, macrolactins, bacillomycin-D, and catechol-based siderophore bacillibactin which were further confirmed by amplifying the genes involved in the biosynthesis of these antimicrobial lipopeptides. When compared for the amounts of different cyclic-peptides produced by three Bacillus strains, B. amyloliquefaciens SB-1 showed the most noticeable amounts of all the antifungal compounds. Plant experiment results revealed that inoculation with phytohormone producing Bacillus spp. strains demonstrated substantial growth improvement of wheat biomass, number of spikes, and dry weight of shoots and roots. Results of this study indicate the biocontrol and biofertilizer potential of Bacillus spp. for sustainable plant nutrient management, growth promotion, and effective biocontrol of crop plants, particularly cultivated in the South Asian region.

Chemical and pharmacological evaluation of the non-flowering aerial parts of Acacia modesta Wall. cultivated in Egypt

BackgroundAcacia modesta Wall. (A. modesta), often recognized as Phulai, is belonging to family Fabaceae and sub-family Mimosaceae. A. modesta has many beneficial uses. Leaves, wood, flowers, and gum of A. modesta have been used frequently for multiple therapeutic purposes.ResultsThe chemical investigation of butanol fraction of A. modesta non-flowering aerial parts yielded Vitexin-2′′-β-D-glucopyranoside and Apigenin-6,8-di-C-β-D-glucopyranoside in a flavone mixture as well as (β-D-glucopyranosyl (1-3)-β-D-glucopyranosyl)-3-β-hydroxy-11-oxo-olean-12-en-28-oic acid) an oleanane-type triterpenoidal saponin. Metabolite profiling via ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) of the ethyl acetate fraction resulted in recognizing of eighteen compounds tentatively compared with previously published data. Quantitative measurement of the overall value of flavonoids of A. modesta was found to be 2.824 μg/100 μg ± 0.01 calculated as quercetin. The acute toxicity study of the ethanol extract proved that the plant under investigation is safe and nontoxic to the male albino mice used. The anti-hyperglycemic activity of the ethanol extract performed on type 2 diabetic rats proved that the most potent dosage was 200 mg/kg b. wt. after 4 and 4 weeks of treatment respectively compared to metformin. Furthermore, evaluation of the hepato-protective activity of the ethanol extract of the plant under investigation showed that the most potent extract was with a dose level of 200 mg/kg b. wt. after 3 and 10 days of continuous treatment compared to silymarin.ConclusionIt can be concluded that A. modesta Wall. cultivated in Egypt could be used as a promising anti-diabetic agent and a hepato-protective agent against hepatocellular damage induced by hepatotoxins.

Coupling Filter-Based Thermal Desorption Chemical Ionization Mass Spectrometry with Liquid Chromatography/Electrospray Ionization Mass Spectrometry for Molecular Analysis of Secondary Organic Aerosol.

Filter-based thermal desorption (F-TD) techniques, such as the filter inlet for gases and aerosols, are widely employed to investigate the molecular composition and physicochemical properties of secondary organic aerosol (SOA). Here, we introduce an enhanced capability of F-TD through the combination of a customized F-TD inlet with chemical ionization mass spectrometry (CIMS) and ultraperformance liquid chromatography/electrospray ionization mass spectrometry (UPLC/ESI-MS). The utility of F-TD/CIMS + UPLC/ESI-MS is demonstrated by application to α-pinene ozonolysis SOA for which increased filter aerosol mass loading is shown to slow the evaporation rates of deposited compounds. Evidence for oligomer decomposition producing multimode F-TD/CIMS thermograms is provided by the measurement of the mass fraction remaining of monomeric and dimeric α-pinene oxidation products on the filter via UPLC/ESI-MS. In situ evaporation of aerosol particles suggests that α-pinene-derived hydroperoxides are thermally labile; thus, analysis of particle-phase (hydro)peroxides via F-TD may not be appropriate. A synthesized pinene-derived dimer ester (C20H32O5) is found to be thermally stable up to 200 °C, whereas particle-phase dimers (C19H30O5) are observed to form during F-TD analysis via thermally induced condensation of synthesized pinene-derived alcohols and diacids. The complementary F-TD/CIMS + UPLC/ESI-MS method offers previously inaccessible insight into the molecular composition and thermal desorption behavior of SOA that both clarifies and expands on analysis via traditional F-TD techniques.

Targeted Development-Dependent Metabolomics Profiling of Bioactive Compounds in Acanthopanax senticosus by UPLC-ESI-MS

An ultra-performance liquid chromatography-electrospray ionization-mass spectrometry targeted metabolomics strategy was applied to analyze protocatechuate, syringin, eleutheroside E, isofraxidin, hyperoside, kaempferol, and oleanolic acid, the active compounds in 3-year-old, 5-year-old, and 9-year-old Acanthopanax senticosus. Then, targeted metabolomics was conducted with 3 growth year plants to identify 19 phenolic metabolites related to the above-mentioned active compounds, including 9 C6C3C6-type, 6 C6C3-type, and 4 C6C1-type. Multivariate statistical analysis was applied to the bioactive metabolite data, and targeted metabolic profiling was used for marker compound classification and characterization. The results showed that 7 active compounds in the roots and stems in the 3 growth year plants differed. The principal component “Q” values showed that the total contents of 7 active compounds in 5-year-old roots and stems were higher than in other growth years. Results of targeted metabolomics profiling of 19 phenolic metabolites showed that the C6C1-type compounds accumulated in 9-year-old plants, the C6C3-type in 3-year-old plants, and the C6C3C6-type in 5-year-old plants. The stems had the greatest accumulations of the phenolic metabolites. C6C1 and C6C3-type metabolites are the most abundant in both roots and stems. In conclusion, the active compounds and pharmacological effects of A. senticosus in different growth years are different. The best harvest age for A. senticosus roots and stems was 5 years. The accumulation of 19 phenolic metabolites in different growth years also showed significant differences.

Changes in endogenous auxin level during flower bud abscission process in Pistachio (Pistacia vera L.)

The regulation of auxin (indole-3-acetic acid, IAA) level in plants is a complex mechanism and the concept is not completely understood. There are many lines of evidence suggesting that amino acid conjugates of IAA play a vital role in auxin homeostasis, but the involved plant enzymes are still under investigation. In the present study, the alteration in the levels of auxin and its conjugates in the different physiological stages and organs, as well as its role in flower bud abscission in Pistacia vera L., were investigated. The levels of the auxin and its conjugates, such as IAA, indole butyric acid (IBA), IAA-Ala, IAA-Leu, IAA-Asp, and IAA-Glu, were analyzed by ultraperformance liquid chromatography-electrospray ionization-mass spectrometry (UPLC/ESI-MS/MS) technique. Among the auxin conjugates, IAA was the main conjugate and dominant indole detected in pistachio extract observed in this work. It has also been determined that quite a few organs of 'Off-year' trees have higher auxin content than 'On-year' trees. It was observed that the auxin level is low in fruit-bearing trees in both leaves and shoots. It affects fruit yield and fruit bud abscission and plays some role in the alternate bearing. Decreased levels of auxin were observed in most of the organs of 'On-year' trees during kernel development resulting in bud abscission. This study shows that auxin conjugates play an important role in IAA metabolism, particularly in flower bud abscission and fruit kernel development.

Momilactones A, B, and Tricin in Rice Grain and By-Products are Potential Skin Aging Inhibitors.

We previously reported the inhibitory potentials of momilactones A (MA) and B (MB) against key enzymes related to type 2 diabetes and obesity. In this study, antioxidant and anti-skin-aging activities of MA and MB were investigated and compared with tricin, a well-known antioxidant and antiaging flavonoid in rice. MA, MB, and tricin were purified from rice husk by column chromatography and their biological activities were subsequently assayed by in vitro trials. The contents of MA, MB, and tricin of different commercial rice cultivars in Japan were quantified and confirmed by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) and high-performance liquid chromatography (HPLC) analyses. The antioxidant assays revealed a synergistic activity of the mixture MA and MB (MAB, 1:1, v/v). In addition, in 2,2’-azino-bis (ABTS) assay, IC50 values of MAB (0.3 mg/mL) and tricin (0.3 mg/mL) was 4-fold and 9-fold greater than that of individual MB (1.3 mg/mL) or MA (2.8 mg/mL), respectively. The in vitro enzymatic assays on pancreatic elastase and tyrosinase indicated that MA and MB were potential to relief skin wrinkles and freckles. In detail, MA exerted higher inhibition on both enzymatic activities (30.9 and 37.6% for elastase and tyrosinase inhibition, respectively) than MB (18.5 and 12.6%) and MAB (32.0 and 19.7%) at a concentration of 2.0 mg/mL. Notably, MA and the mixture MAB exhibited stronger inhibitions on elastase and tyrosinase in comparison with tricin and vanillin. MA, MB, and tricin in rice are potential to develop cosmetics as well as supplements for skin aging treatments.

Analysis of Phenolic Composition of Byproducts (Seeds and Peels) of Avocado (Persea americana Mill.) Cultivated in Colombia.

The byproducts (seeds and peels) of an avocado cultivated in the south of Colombia were extracted with aqueous acetone and their antioxidant properties were measured with ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) assays, and total polyphenol content was determined by Folin–Ciocalteu method. A bioguided fractionation was performed, first by SPE (solid phase extraction) on Amberlite XAD-7, and then by size exclusion chromatography on Sephadex LH-20. The polyphenolic-rich extracts and their fractions were analyzed by ultra-performance liquid chromatography–electrospray ionization–mass spectrometry (UPLC-ESI-MS/MS), finding the presence of organic acids, hydroxycinnamic acids, catechins, free and glycosylated flavonoids, and dimeric and trimeric procyanidins. Catechin, epicatechin, six quercetin derivatives, four dimeric procyanidins (three type B and one type A), and three trimeric procyanidins (two type B and one type A) were detected in the most active fractions of avocado peel and seeds. The most antioxidant fractions contain the higher molecular weight phenolic compounds (condensed tannins).

Simultaneous quantitation of aconitum alkaloids from You-Gui-yin in rat plasma by UPLC–ESI–MS and its application to a pharmacokinetic study

You-Gui-Yin (YGY), a famous traditional Chinese medicine, has been widely used in clinics for the treatment of kidney-yang deficiency, yang deficiency caused by excessive yin, and osteoporosis. A rapid and sensitive ultraperformance liquid chromatography–electrospray ionization–mass spectrometry (UPLC–ESI–MS) method for simultaneous determination of six Aconitum alkaloids including aconitine (AC), hypaconitine (HA), mesaconitine (MA), benzoylaconine (BAC), benzoylhypaconine (BHA), and benzoylmesaconine (BMA) in rat plasma after oral administration of YGY was developed in this study. Chromatographic separation was performed on an ACQUITY UPLC™ BEH C18 column (2.1 × 100 mm, 1.7 μm) using gradient elution with the mobile phase consisting of 2 mmol/L ammonium formate in 0.05% formic acid aqueous solution and 0.05% formic acid methanol solution, at a flow rate of 0.20 mL/min. MS detection was performed in the positive ion mode. The calibration curves were linear in the concentration range of 0.04160–41.60 ng/mL, 0.1070–107.0 ng/mL, 0.07358–73.50 ng/mL, 0.03228–32.28 ng/mL, 0.01809–18.09 ng/mL, and 0.1320–132.0 ng/mL for AC, HA, MA, BAC, BHA, and BMA, respectively. The intra- and inter-day precisions (relative standard deviation [RSD]) were less than 11.6% and 12.6%, respectively. The accuracies Relative Error (RE) ranged from −10.2% to 5.6%, while the recoveries ranged from 70.4% to 99.3%. The method for simultaneous quantitation of Aconitum alkaloids of You-Gui-Yin in rat plasma is accurate and repeatable, and this method was successfully applied to investigate the pharmacokinetics of the six Aconitum alkaloids in rat plasma after oral administration of YGY. For the pharmacokinetic study, the pharmacokinetics of the six Aconitum alkaloids were best described by a two-compartment open model.

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.

Isolation and identification of a novel algicidal peptide from mackerel muscle hydrolysate

To help remedy damage from harmful algal blooms, an attempt was made to isolate an algicidal substance previously observed to be present in mackerel muscle hydrolysate. Crude extract was obtained by cold acetone precipitation, and it dissolved best in water. Through molecular weight cut-off determination and tricine-SDS PAGE, the algicidal substance was determined to be a peptide of <1 kDa. Based on this result, purification was first performed using size exclusion chromatography and preparative reverse phase high-performance liquid chromatography. Then, the active algicidal fraction was applied to an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry system, followed by MS/MS analysis. The algicidal peptide had linear structure consisting of amino acids with sequence NH-KMNF-COOH. Its calculated properties were: molecular weight 538.66 g/mol; isoelectric point 9.91; net charge +1 at pH 7.0; and 50% hydrophobicity. Algicidal ability of the identified peptide was confirmed using synthesized peptide. The LC50 values toward four harmful algal blooming species were 0.69, 0.83, 0.85 and 1.24 mg/ml for Alexandrium fundyense, A. catenella, Heterocapsa triquetra, and Prorocentrum minimum, respectively. There was no coincidence in the sequence of the identified peptide with those of known metabolites in the APD, Norine, CAMP, UniProt and METLIN databases. Consequently, this algicidal substance originating from mackerel protein was deduced to be a novel peptide that can usefully be applied to relieve harmful algal blooms.

Glycyrrhetinic Acid Accelerates the Clearance of Triptolide through P-gp In Vitro.

Triptolide (TP) is an active ingredient isolated from Tripterygium wilfordii Hook. f. (TWHF), which is a traditional herbal medicine widely used for the treatment of rheumatoid arthritis and autoimmune disease in the clinic. However, its adverse reactions of hepatotoxicity and nephrotoxicity have been frequently reported which limited its clinical application. The aim of this study was to investigate the mechanism of glycyrrhetinic acid (GA) effecting on the elimination of TP in HK-2 cells and the role of the efflux transporters of P-gp and multidrug resistance-associated proteins (MRPs) in this process. An ultra performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) analytical method was established to determine the intracellular concentration of TP. In order to study the role of efflux transporters of P-gp and MRPs in GA impacting on the accumulation of TP, the inhibitors of efflux transporters (P-gp: verapamil; MRPs: MK571) were used in this study. The results showed that GA could enhance the elimination of TP and reduce the TP accumulation in HK-2 cells. Verapamil and MK571 could increase the intracellular concentration of TP; in addition, GA co-incubation with verapamil significantly increased the TP cellular concentration compared with the control group. In conclusion, GA could reduce the accumulation of TP in HK-2 cells, which was related to P-gp. This is probably one of the mechanisms that TP combined with GA to detoxify its toxicity. Copyright © 2017 John Wiley & Sons, Ltd.

Determination of S-methyl-L-methionine (SMM) from Brassicaceae Family Vegetables and Characterization of the Intestinal Transport of SMM by Caco-2 Cells.

The objectives of the current study were to determine S-methyl-L-methionine (SMM) from various Brassicaceae family vegetables by using validated analytical method and to characterize the intestinal transport mechanism of SMM by the Caco-2 cells. The SMM is well known to provide therapeutic activity in peptic ulcers. The amount of SMM from various Brassicaceae family vegetables ranged from 89.08 ± 1.68 μg/g to 535.98 ± 4.85 μg/g of dry weight by using validated ultra-performance liquid chromatography-electrospray ionization-mass spectrometry method. For elucidating intestinal transport mechanism, the cells were incubated with or without transport inhibitors, energy source, or a metabolic inhibitor. Phloridzin and verapamil as inhibitors of sodium glucose transport protein (SGLT1) and P-glycoprotein, respectively, were not responsible for cellular uptake of SMM. Glucose and sodium azide were not affected by the cellular accumulation of SMM. The efflux ratio of SMM was 0.26, implying that it is not effluxed through Caco-2 cells. The apparent coefficient permeability (Papp ) of SMM was 4.69 × 10-5 cm/s, indicating that it will show good oral absorption in in vivo.

Simultaneous determination of anemoside B4, phellodendrine, berberine, palmatine, obakunone, esculin, esculetin in rat plasma by UPLC–ESI–MS/MS and its application to a comparative pharmacokinetic study in normal and ulcerative colitis rats

A sensitive and rapid ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC–ESI–MS/MS) method was developed for the simultaneous analysis of anemoside B4, phellodendrine, berberine, palmatine, obakunone, esculin, esculetin, toosendanin (IS1 of anemoside B4), tetrahydropalmatine (IS2 of phellodendrine, berberine, palmatine and obakunone) and scopoletin (IS3 of esculin and esculetin) and to compare the pharmacokinetics of these active ingredients in normal and ulcerative colitis rats. After methanol deproteinization, solvents were evaporated at 40°C under a gentle stream of nitrogen. Chromatography was performed using a C18 column with a gradient elution of 0.1% aqueous formic acid and acetonitrile at 0.4ml/min. Detection and measurement were performed on a 4000 QTRAP UPLC–MS/MS system from AB Sciex in the multiple reaction monitoring (MRM) mode. Phellodendrine, berberine, palmatine, obakunone, esculin, esculetin, tetrahydropalmatine (IS2) and scopoletin (IS3) were monitored under positive ionization conditions. Anemoside B4, and toosendanin (IS1) were monitored under negative ionization conditions. The optimized mass transition ion-pairs (m/z) were 1221.1/750.7 for anemoside B4, 343.2/193.2 for phellodendrine, 337.1/321.0 for berberine, 353.0/336.9 for palmatine, 455.1/161.1 for obakunone, 341.2/179.2 for esculin, 179.1/123.0 for esculetin, 573.4/531.4 for toosendanin (IS1), 356.2/192.2 for tetrahydropalmatine (IS2) and 193.0/133.1 for scopoletin (IS3).

Profiling of ginsenosides in the two medicinal Panax herbs based on ultra-performance liquid chromatography-electrospray ionization-mass spectrometry.

As the king of herb plants, ginseng has been used for nearly 5000 years in medicines in Asia and recently in the West. Ginsenosides, the main active constituents in Panax herbs, have prominent immunoregulatory effects. Although extensively studied in the roots, ginsenosides have not been studied with regard to their profiles and natural variations in the leaf, stem, petiole, lateral root, and main roots during development or among species. In this study, a sensitive ultra-performance liquid chromatography-electrospray ionization–mass spectrometry method with a shorter chromatographic running time was developed and validated for simultaneous quantification of ten ginsenosides. Comparing ginsenoside contents in various parts during different developmental stages revealed part-specific accumulation of most ginsenosides. Further investigation indicated that Rg3 accumulated at significantly higher levels in the petiole of P. ginseng than in that of P. quinquefolius. The relative ratio of ginsenoside Rb2 to Rb1 appears to be a candidate metabolic marker for identifying the ginseng cultivar within a diverse collection of ginseng accessions. In addition, the PCA showed that aboveground parts differed significantly between species and can be considered as species-specific markers rather than roots. This comprehensive survey, providing reliable, affordable and adequate scientific evidence, could be used to differentiate two species and discriminate ginseng cultivar ages.Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-016-3427-3) contains supplementary material, which is available to authorized users.

Traditional herbal prescription LASAP-C inhibits melanin synthesis in B16F10 melanoma cells and zebrafish.

BackgroundIn this study, the anti-melanogenesis efficacy of clinically used herbal prescription LASAP-C, which consists of four herbal medicines—Rehmanniae Radix Crudus, Lycii Fructus, Scutellariae Radix, and Angelicae Dahuricae Radix, was investigated.MethodsThe chemical profile of LASAP-C was established by conducting ultra-performance liquid chromatography-electrospray ionization-mass spectrometry. Anti-melanogenic efficacy was evaluated by tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 expression in B16F10 melanoma cells. In vivo evaluation was performed by using zebrafish model.ResultsMolecular evidences suggested that melanin synthesis was inhibited via the down-regulation of tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 expression in B16F10 melanoma cells treated with LASAP-C. The anti-melanogenesis efficacy was also confirmed in vivo by using the zebrafish model.ConclusionThe results of this study provide strong evidences that LASAP-C can be used as an active component in cosmeceutical products for reducing excess pigmentation in the human skin.