Improving Bioaccessibility and Bioavailability of Isoflavone Aglycones from Chickpeas by Germination and Forming ß-Cyclodextrin Inclusion Complexes.

Chickpea isoflavones have diverse pharmacological activities but with low water solubility and bioavailability. In this work, the isoflavone content in chickpeas was first increased by germination, and then the bioaccessibility and bioavailability of isoflavones in chickpea sprout extracts (CSE) were enhanced using ß-cyclodextrin (ß-CD) inclusion techniques. Firstly, the total content of isoflavones was increased by 182 times through sprouting, and isoflavones were presented mostly in the germ and radicle. Then, the chickpea sprout extract/ß-cyclodextrin (CSE/ß-CD) inclusion complex was prepared and characterized. The in vitro test showed that the cumulative release of two isoflavones, formononetin (FMN) and biochanin A (BCA), in the CSE/ß-CD was significantly increased in a simulated digestive fluid. The in vivo rat pharmacokinetics demonstrated that the inclusion of FMN and BCA by ß-CD effectively increased their bioavailability in rat plasma and tissues, especially in the liver. The study provides a feasible strategy for improving the bioavailability of isoflavones from chickpeas and is also beneficial to the utilization of other legume resources.

Integrating compound-specific stable isotope and enantiomer-specific analysis to characterize the isomeric and enantiomeric signatures of hexachlorocyclohexanes (HCHs) in paddy soils.

Organic pollutants in paddy fields may undergo different processes from those in dryland due to the anaerobic environment. The integrated use of compound-specific stable isotope analysis (CSIA) and enantiomer-specific analysis is a promising technique for understanding the behavior and fate of organic pollutants in soils. In this study, soil samples were collected from paddy fields in three major rice cultivation regions of China, spanning a transect of 4000 km. The mean concentrations of Æ©HCHs in paddy soils from the Taihu Plain were the highest (1.44 ng/g). The ratios of α-HCH/ß-HCH (all below 11.8) and α-HCH/γ-HCH (92% below 4.64), as well as the enantiomeric fractions (EFs) of chiral α-HCH (mean of 0.81), reflected that the distribution of HCHs was affected by the use of both technical HCHs and lindane. The preferential depletion of (-)-α-HCH and pronounced carbon isotope fractionation of α-HCH (δ13C of -28.22 ± 0.92‰ -23.63 ± 1.89‰) demonstrated its effective transformation. Factors such as altitude, soil temperature, soil pH, soil conductivity and soil organic matter significantly influenced the fate and transformation of HCHs. The current study highlights the integrated application of CSIA and enantiomer-specific analysis to provide multiple lines of evidence for the transformation of HCHs in soils.

Low-protein diet supplemented with inulin lowers protein-bound toxin levels in patients with stage 3b-5 chronic kidney disease: a randomized controlled study / La dieta baja en proteínas suplementada con inulina reduce los niveles de toxinas unidas a proteínas en pacientes con enfermedad renal crónica en estadio 3b-5: un estudio controlado aleatorio

Objective: this study aimed to evaluate whether low-salt low-protein diet (LPD) supplemented with 10 g of inulin could lower serum toxin levels in patients with chronic kidney disease (CKD), thereby providing evidence for adjusting dietary prescriptions of inhospital patients and outpatient nutrition consultants. Methods: we randomized 54 patients with CKD into two groups. Dietary protein intake compliance was evaluated using a 3-day dietary diary and 24-h urine nitrogen levels. The primary outcomes were indoxyl sulfate (IS) and p-cresyl sulfate (PCS), and secondary outcomes included inflammation marker levels, nutritional status, and renal function. We assessed 89 patients for eligibility, and a total of 45 patients completed the study, including 23 and 22 in the inulin-added and control groups, respectively. Results: PCS values decreased in both groups after intervention: inulin-added group, ∆PCS -1.33 (-4.88, -0.63) μg/mL vs. LPD group, -4.7 (-3.78, 3.69) μg/mL (p = 0.058). PCS values reduced from 7.52 to 4.02 μg/mL (p < 0.001) in the inulin-added group (p < 0.001). Moreover, IS decreased from 3.42 (2.53, 6.01) μg/mL to 2.83 (1.67, 4.74) μg/mL after adding inulin; ∆IS was -0.64 (-1.48, 0.00) μg/mL, and a significant difference was observed compared with the control group (p = 0.004). The inflammation index decreased after intervention. Conclusion: dietary fiber supplementation may reduce serum IS and PCS levels and modulate their inflammatory status in predialysis CKD patients.(AU)

Objetivo: este ensayo aleatorizado doble ciego comparó el efecto de una dieta baja en proteínas (LPD) con o sin suplementos orales de 10 gde inulina en los niveles de PBUT en pacientes con ERC en prediálisis durante 12 semanas.Métodos: clasificamos aleatoriamente a 54 pacientes con ERC en dos grupos. El cumplimiento de la ingesta dietética de proteínas se evaluóutilizando un diario dietético de 3 días y nitrógeno en orina de 24 horas. Los resultados primarios fueron IS y PCS y los resultados secundariosincluyeron niveles de marcadores de inflamación, estado nutricional y función renal. Evaluamos la elegibilidad de 89 pacientes y 45 completaronla intervención, incluidos 23 y 22 en los grupos de inulina añadida y de control, respectivamente.Resultados: el sodio urinario promedio de 24 horas fue de 86 mmol/día y la ingesta promedio de proteínas fue de ~0,7 g/kg/día. Los valores dePCS exhibieron una tendencia decreciente en ambos grupos después de la intervención: grupo con inulina añadida, ∆PCS -1.33 (-4.88, -0.63)μg/mL vs. grupo LPD, -4.7 (-3.78, 3.69) μg/mL) (p =0,058). Los valores de PCS se redujeron de 7,52 a 4,02 μg/mL (p < 0,001) con inulina(p < 0,001). Además, IS disminuyó de 3,42 (2,53, 6,01) μg/mL a 2,83 (1,67, 4,74) μg/mL después de agregar inulina; El ∆IS fue -0,64 (-1,48;0,00) μg/mL y se observó una diferencia significativa en comparación con el grupo control (p =0,004).Conclusión: la suplementación con fibra dietética puede reducir las toxinas de unión a proteínas séricas en pacientes con ERC en prediálisisy modular su estado inflamatorio.(AU)

Low-protein diet supplemented with inulin lowers protein-bound toxin levels in patients with stage 3b-5 chronic kidney disease: a randomized controlled study.

Introduction: Objective: this study aimed to evaluate whether low-salt low-protein diet (LPD) supplemented with 10 g of inulin could lower serum toxin levels in patients with chronic kidney disease (CKD), thereby providing evidence for adjusting dietary prescriptions of inhospital patients and outpatient nutrition consultants. Methods: we randomized 54 patients with CKD into two groups. Dietary protein intake compliance was evaluated using a 3-day dietary diary and 24-h urine nitrogen levels. The primary outcomes were indoxyl sulfate (IS) and p-cresyl sulfate (PCS), and secondary outcomes included inflammation marker levels, nutritional status, and renal function. We assessed 89 patients for eligibility, and a total of 45 patients completed the study, including 23 and 22 in the inulin-added and control groups, respectively. Results: PCS values decreased in both groups after intervention: inulin-added group, ∆PCS -1.33 (-4.88, -0.63) µg/mL vs. LPD group, -4.7 (-3.78, 3.69) µg/mL (p = 0.058). PCS values reduced from 7.52 to 4.02 µg/mL (p < 0.001) in the inulin-added group (p < 0.001). Moreover, IS decreased from 3.42 (2.53, 6.01) µg/mL to 2.83 (1.67, 4.74) µg/mL after adding inulin; ∆IS was -0.64 (-1.48, 0.00) µg/mL, and a significant difference was observed compared with the control group (p = 0.004). The inflammation index decreased after intervention. Conclusion: dietary fiber supplementation may reduce serum IS and PCS levels and modulate their inflammatory status in predialysis CKD patients.

Introducción: Objetivo: este ensayo aleatorizado doble ciego comparó el efecto de una dieta baja en proteínas (LPD) con o sin suplementos orales de 10 g de inulina en los niveles de PBUT en pacientes con ERC en prediálisis durante 12 semanas. Métodos: clasificamos aleatoriamente a 54 pacientes con ERC en dos grupos. El cumplimiento de la ingesta dietética de proteínas se evaluó utilizando un diario dietético de 3 días y nitrógeno en orina de 24 horas. Los resultados primarios fueron IS y PCS y los resultados secundarios incluyeron niveles de marcadores de inflamación, estado nutricional y función renal. Evaluamos la elegibilidad de 89 pacientes y 45 completaron la intervención, incluidos 23 y 22 en los grupos de inulina añadida y de control, respectivamente. Resultados: el sodio urinario promedio de 24 horas fue de 86 mmol/día y la ingesta promedio de proteínas fue de ~0,7 g/kg/día. Los valores de PCS exhibieron una tendencia decreciente en ambos grupos después de la intervención: grupo con inulina añadida, ∆PCS -1.33 (-4.88, -0.63) µg/mL vs. grupo LPD, -4.7 (-3.78, 3.69) µg/mL) (p =0,058). Los valores de PCS se redujeron de 7,52 a 4,02 µg/mL (p < 0,001) con inulina (p < 0,001). Además, IS disminuyó de 3,42 (2,53, 6,01) µg/mL a 2,83 (1,67, 4,74) µg/mL después de agregar inulina; El ∆IS fue -0,64 (-1,48; 0,00) µg/mL y se observó una diferencia significativa en comparación con el grupo control (p =0,004). Conclusión: la suplementación con fibra dietética puede reducir las toxinas de unión a proteínas séricas en pacientes con ERC en prediálisis y modular su estado inflamatorio.

NMR-Based Metabolomics to Analyze the Effects of a Series of Monoamine Oxidases-B Inhibitors on U251 Cells.

Alzheimer's disease (AD) is a typical progressive neurodegenerative disorder, and with multiple possible pathogenesis. Among them, coumarin derivatives could be used as potential drugs as monoamine oxidase-B (MAO-B) inhibitors. Our lab has designed and synthesized coumarin derivatives based on MAO-B. In this study, we used nuclear magnetic resonance (NMR)-based metabolomics to accelerate the pharmacodynamic evaluation of candidate drugs for coumarin derivative research and development. We detailed alterations in the metabolic profiles of nerve cells with various coumarin derivatives. In total, we identified 58 metabolites and calculated their relative concentrations in U251 cells. In the meantime, the outcomes of multivariate statistical analysis showed that when twelve coumarin compounds were treated with U251cells, the metabolic phenotypes were distinct. In the treatment of different coumarin derivatives, there several metabolic pathways changed, including aminoacyl-tRNA biosynthesis, D-glutamine and D-glutamate metabolism, glycine, serine and threonine metabolism, taurine and hypotaurine metabolism, arginine biosynthesis, alanine, aspartate and glutamate metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, glutathione metabolism and valine, leucine and isoleucine biosynthesis. Our work documented how our coumarin derivatives affected the metabolic phenotype of nerve cells in vitro. We believe that these NMR-based metabolomics might accelerate the process of drug research in vitro and in vivo.

Components research on Tetrastigma hemsleyanum Diels et Gilg: Identification and effect of drying methods on the content of ten main constituents by targeting metabolomics method.

Tetrastigma hemsleyanum Diels et Gilg (TH) is one of the new eight Genuine Medicinal Materials of Zhejiang. It has extensive biological activities, such as anti-inflammatory, anti-tumor and analgesic activities, etc. In this study, the chemical components of TH were systematically investigated by ultra high-performance liquid chromatography-tandem quadrupole time of flight mass spectrometry (UPLC/Q-TOF-MS). Based on the MS spectrum, 39 compounds in TH extracts including 14 flavonoids, 10 fatty acids, 5 polyphenols and phenolic acids, 4 terpenes and other compounds were detected and tentatively identified. TH samples were treated under different drying methods (vacuum freeze drying, hot air drying, natural drying, light drying and vacuum drying). Besides, the effect of different drying methods on the content of 10 main chemical constituents in TH extracts including catechin, rutin, kaempferol-3-O-rutinoside and so on was also investigated by targeting metabolomics method with ultra high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (UPLC-MS/MS) assisted by multivariate statistical analysis. Large differences were observed between vacuum drying and vacuum freeze drying with remarkable content changes. The contents of rutin, proanthocyanidin B1 and catechin were the most different among the various drying methods. The systematic identification of chemical constituents is helpful for the further medicinal development and application of TH. The effects of drying methods on the content of TH components were studied, which provided experimental data for the processing, storage and quality control of TH.

Mechanochemical preparation of red clover extract/ß-cyclodextrin dispersion: Enhanced water solubility and activities in alleviating high-fat diet-induced lipid accumulation and gut microbiota dysbiosis in mice.

Red clover (RC) extract is rich in isoflavones (formononetin and biochanin A) that have various biological functions. However, its low water solubility limits its bioavailability. In this study, an RC extract/ß-cyclodextrin (RC/ß-CD) dispersion was prepared by ball milling to enhance its water solubility and biological availability. The water solubility of formononetin and biochanin A was 34.45 and 13.65 µg/mL (increased to 3.11 and 2.14 times higher than that of RC alone), respectively. The alleviating effects of the dispersion on lipid accumulation and gut microbiota were evaluated in mice. The RC/ß-CD dispersion showed a better effect on inhibiting lipid accumulation, especially on total triglycerides. The dispersion group had a higher relative abundance of Akkermansia, Muribaculaceae, and Bacteroides than RC alone, along with a higher level of acetic and butyric acid. The study provides a feasible way for improving the bioaccessibility and bioactivity of RC isoflavones in red clover.

Study on Absorption, Distribution and Excretion of a New Candidate Compound XYY-CP1106 against Alzheimer's Disease in Rats by LC-MS/MS.

XYY-CP1106, a candidate compound synthesized from a hybrid of hydroxypyridinone and coumarin, has been shown to be remarkably effective in treating Alzheimer's disease. A simple, rapid and accurate high-performance liquid chromatography coupled with the triple quadrupole mass spectrometer (LC-MS/MS) method was established in this study to elucidate the pharmacokinetics of XYY-CP1106 after oral and intravenous administration in rats. XYY-CP1106 was shown to be rapidly absorbed into the blood (Tmax, 0.57-0.93 h) and then eliminated slowly (T1/2, 8.26-10.06 h). Oral bioavailability of XYY-CP1106 was (10.70 ± 1.72)%. XYY-CP1106 could pass through the blood-brain barrier with a high content of (500.52 ± 260.12) ng/g at 2 h in brain tissue. The excretion results showed that XYY-CP1106 was mainly excreted through feces, with an average total excretion rate of (31.14 ± 0.05)% in 72 h. In conclusion, the absorption, distribution and excretion of XYY-CP1106 in rats provided a theoretical basis for subsequent preclinical studies.

Investigation on Antioxidant Activity and Different Metabolites of Mulberry (Morus spp.) Leaves Depending on the Harvest Months by UPLC-Q-TOF-MS with Multivariate Tools.

The changes in active components in mulberry leaves harvested in different months and their antioxidant activities were investigated. Ultra-high-performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) with multivariate statistical tools was used to investigate the chemical constituents in the extracts of mulberry leaves. The results indicated that mulberry leaves were rich in phenolic acids, flavonoids, organic acids, and fatty acid derivatives. In addition, 25 different compounds were identified in the different batches of mulberry leaves. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured to evaluate the in vitro antioxidant activities of mulberry leaves. Among the four batches, batch A, harvested in December, exhibited the strongest DPPH radical-scavenging activity, while batch B, harvested in March, showed the weakest activity. This was related to the total phenolic content in the mulberry leaves of each batch. The optimal harvest time of mulberry leaves greatly influences the bioactivity and bioavailability of the plant.

Metabolomics analysis of differential chemical constituents and α-glucosidase inhibiting activity of Phyllanthus urinaria L. root, stem, leaf and fruit.

The differential chemical constituents of the different Phyllanthus urinaria L. (PUL) parts were investigated by UPLC/Q-TOF MS-based metabolomics. A total of 69 compounds were tentatively identified in the whole plant extract and 35 of them were common to root, stem, leaf and fruit parts. And four compounds were selected as biomarkers for leaves, fruits, stems and roots, respectively. The four PUL parts all had good α-glucosidase inhibitory activities and the activities of fruit, root and stem extracts were about fivefold higher than the leaf part. The hierarchical cluster analysis and heat map were used to explore the relationship between the α-glucosidase inhibitory activities and chemical constituent differences of four PUL parts.

Study of Chemical Compositions and Anticancer Effects of Paris polyphylla var. Chinensis Leaves.

Paris polyphylla var. chinensis (Franch.) Hara is a perennial herb belonging to the Trilliaceae family. Ultraperformance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) was used to detect the composition of different fractions of Paris polyphylla var. chinensis leaves. Meanwhile, the extracts of different fractions were evaluated for their cytotoxic activities against four selected human cancer cell lines and one human normal epithelial cell line based on the MTT assay method. Multivariate statistical analysis was performed to screen differential compounds and to analyze the distributions between different fractions. Finally, more than 60 compounds were obtained and identified from the different fractions of Paris polyphylla var. chinensis leaves, and the chloroform and n-butanol extracts showed significant cytotoxic effects on these four cancer cells. Several compounds were preliminarily identified from different fractions, including 36 steroidal saponins, 11 flavonoids, 10 ceramides, 8 lipids, 6 organic acids, and 8 other compounds. Various compounds were screened out as different chemical components of different fractions, which were considered as a potential substance basis for the cytotoxicity of Paris polyphylla var. chinensis leaves.

Mechanochemical-Assisted Extraction and Hepatoprotective Activity Research of Flavonoids from Sea Buckthorn (Hippophaë rhamnoides L.) Pomaces.

Pomaces of sea buckthorn berry were usually side-products during the processing of juice. Due to a lack of an economical and effective extraction method, it was typically recognized as waste. For the purpose of resource utilization, the mechanochemical-assisted extraction (MCAE) method was applied to develop an ecofriendly extraction method and product with better pharmacology activity. The parameters were investigated through response surface methodology (RSM) design experiments. The processing conditions were optimized as follows: amount of Na2CO3 40%, ball-to-material rate 29:1 g/g, milling speed 410 rpm, milling time 24 min, extraction temperature 25 °C, extraction time 20 min and the solid-to-solution ratio 1:10 g/mL. Under these conditions, the yields of flavonoids from sea buckthorn pomaces were 26.82 ± 0.53 mg/g, which corresponds to an increase of 2 times in comparison with that extracted by the heat reflux extraction method. Meanwhile, the hepatoprotective activity of sea buckthorn pomaces extracts was studied by the liver injury induced by ip injection of tetracycline. Biochemical and histopathological studies showed that biomarkers in serum and liver of nonalcoholic fatty liver disease (NAFLD) mice were significantly ameliorated when sea buckthorn flavonoids extracted by MCAE were used. Altogether, these results demonstrate that, as a green and efficient extraction, MCAE treatment could increase the extraction yield of sea buckthorn flavonoids, meanwhile it could exhibit significant activity of improving liver function. This research provided a new way to use pomaces of sea buckthorn as a functional food. It also has great value on the comprehensive utilization of nature's resources.

Effect of Drying Methods on Volatile Compounds of Burdock (Arctium lappa L.) Root Tea as Revealed by Gas Chromatography Mass Spectrometry-Based Metabolomics.

Burdock (Arctium lappa L.) is one of the nutritional foods widely planted in many countries. Dried burdock root (BR) is available as a herbal tincture and tea in many Asian countries with good flavor and taste. In this study, the volatile components in dried BR were identified and the effects of different drying methods on the volatile components were investigated by HS-GC-MS method. A total of 49 compounds were identified. Different drying methods including hot-air drying (HD, at 50, 60, 70, and 80 °C), vacuum drying (VD, at 50, 60, 70, and 80 °C), sunlight drying (SD), natural drying (ND), and vacuum freeze drying (VFD) were evaluated by HS-GC-MS-based metabolomics method. Results showed that different drying methods produced different effects on the volatile compounds. It was observed that 2,3-pentanedione, 1-(1H-pyrrol-2-yl)-ethanone, furfural, and heptanal were detected at higher concentrations in HD 80 and VD 70. The traditional HD and SD methods produced more flavor substances than VFD. The BR treated by the VFD method could maintain the shape of the fresh BR pieces while HD50 and VD80 methods could maintain the color of fresh BR pieces. These findings could help better understand the flavor of the corresponding processed BR and provide a guide for the drying and processing of BR tea.

Components identification and isomers differentiation in pigeon pea (Cajanus cajan L.) leaves by LC-MS.

A valid and reliable method based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry using electrospray ionization was established to identify chemical constituents in the ethanol extract of pigeon pea leaves. A total of 58 compounds were detected both in positive and negative modes. Among them, 42 compounds, including 16 flavones, 1 flavonol, 5 flavanones, 9 isoflavones, 1 coumarin, 1 lactone, 6 stilbenes, 2 chalcones, and 1 other compound, were unambiguously identified or tentatively assigned in view of the retention time, the molecular formula, as well as the fragmentation patterns. Moreover, eight sets of isomers were differentiated by the ion trap mass spectrometry based on the fragment ion differences or the abundance differences of the same fragment ions. The energy-resolved mass spectrometry in light of the relative abundance of characteristic fragment ions was adopted in the study.

Components characterisation of Berchemia lineata (L.) DC. by UPLC-QTOF-MS/MS and its metabolism with human liver microsomes.

Investigations were performed on the determination of the main components in Berchemia lineata (L.) DC. (BL) and its metabolism with human liver microsomes (HLM). A total of 35 compounds were detected in BL extracts and 25 of them including 6 naphthopyrones, 10 flavonoids, 2 phenolic acids, 2 phenols, 4 fatty acids and 1 quinone were unambiguously or tentatively identified by UPLC-QTOF-MS/MS. Among them, naphthopyrones were first identified in BL extracts and labelled in chromatography. In addition, the weak inhibitory effects of BL extracts (IC50＝149.25 µg/mL) and rubrofusarin-6-O-α-L-rhamnosyl-(1-6)-O-ß-D-glu-copyranside (the main component of BL extracts, M0; IC50＝82.14 µM) on CYP3A4 were also proved using testosterone as specific probe drug. The main metabolic pathway of M0 by HLM was hydroxylation in its aglycone, the metabolite was tentatively identified as 10-hydroxy-rubrofusarin-6-O-α-L-rhamnosyl-(1-6)-O-ß-D-glucopyranside. Components characterisation and the metabolism with HLM could help the further development and application of BL.

Biochar improves heavy metal passivation during wet anaerobic digestion of pig manure.

Anaerobic digestion (AD) is regarded as an effective treatment to stabilize organic materials and recycle the energy in pig manure. In this study, 0%, 3%, 5%, and 7% biochar (based on dry weight) were added to pig manure to investigate its influence on improving biogas production and reducing heavy metal bioavailability. The potential ecological risk of heavy metals (namely Mn, Zn, Cu, Ni, As, Cd, Pb, and Cr) in digestates was also assessed. Results show that the methane yield was significantly (P < 0.05) increased by 26.7%, 23.0%, and 26.4% following addition of 3%, 5%, and 7% biochar, respectively. Moreover, there was a significant change in the heavy metal speciation in amendment each group. The 5% biochar group showed the highest passivation rate of Ni, As, and Pb, while the highest passivation rate of Cd, Cr, Mn, and Zn was observed with 7% biochar. Although the anaerobic digestion process slightly increased the ecological risk of heavy metals, all tested digestates were still classified as a moderate risk. Results of this study can provide a reference for the treatment of heavy metal pollution in large- and medium-sized anaerobic digesters treating pig manure.

Design, synthesis and biological evaluation of potential anti-AD hybrids with monoamine oxidase B inhibitory and iron-chelating effects.

A series of active hybrids combining 3-hydroxypyridin-4(1H)-one and coumarin pharmacophores were designed and synthesized as potential agents for the treatment of Alzheimer's disease (AD). All the compounds exhibited excellent iron-chelating activities (pFe3+ = 14.8-19.2) and showed favorable monoamine oxidase B (MAO-B) inhibitory effects compared to the reference drug Pargyline (IC50 = 86.9 nM). Among them, compound 11 g displayed the best MAO-B inhibitory activity with an IC50 value of 99.3 nM. Molecular docking analysis showed that compound 11 g could enter the entrance cavity and substrate cavity of MAO-B. Furthermore, the compound 11 g had an excellent antioxidant effect and was capable of protecting from the amyloid-ß1-42 (Aß1-42) induced PC12 cell damage. In silico tools were applied for predicting the blood-brain barrier (BBB) penetration and compound 11 g was proved to overcome the brain exposure challenge. In the mice behavioral study, compound 11 g significantly ameliorated cognitive impairment induced by Scopolamine. More importantly, compound 11 g displayed favorable pharmacokinetic profiles in a rat model. In summary, compound 11 g, with both anti-MAO-B and iron-chelating ability, was proved to be a promising potential anti-AD agent for further optimization.

Fragmentation pathways of deprotonated amide-sulfonamide CXCR4 inhibitors investigated by ESI-IT-MSn , ESI-Q-TOF-MS/MS and DFT calculations.

Amide-sulfonamides provide a potent anti-inflammatory scaffold targeting the CXCR4 receptor. A series of novel amide-sulfonamide derivatives were investigated for their gas-phase fragmentation behaviors using electrospray ionization ion trap mass spectrometry and quadrupole time-of-flight mass spectrometry in negative ion mode. Upon collision-induced dissociation (CID), deprotonated amide-sulfonamides mainly underwent either an elimination of the amine to form the sulfonyl anion and amide anion or a benzoylamide derivative to provide sulfonamide anion bearing respective substituent groups. Based on the characteristic fragment ions and the deuterium-hydrogen exchange experiments, three possible fragmentation mechanisms corresponding to ion-neutral complexes including [sulfonyl anion/amine] complex (INC-1), [sulfonamide anion/benzoylamide derivative] complex (INC-2) and [amide anion/sulfonamide] complex (INC-3), respectively, were proposed. These three ion-neutral complexes might be produced by the cleavages of S-N and C-N bond from the amide-sulfonamides, which generated the sulfonyl anion (Route 1), sulfonamide anion (Route 2) and the amide anion (Route 3). DFT calculations suggested that Route 1, which generated the sulfonyl anion (ion c) is more favorable. In addition, the elimination of SO2 through a three-membered-ring transition state followed by the formation of C-N was observed for all the amide-sulfonamides.

Simultaneous Quantification and Pharmacokinetic Study of Nine Bioactive Components of Orthosiphon stamineus Benth. Extract in Rat Plasma by UHPLC-MS/MS.

Orthosiphon stamineus Benth. (OS) is a traditional folk medicine for the treatment of kidney stones and other urinary tract diseases. In this study, a rapid and sensitive Ultra high-performance liquid chromatography (UHPLC)-MS/MS approach was established and validated for the simultaneous quantification of nine bioactive components in rat plasma. The nine components from OS extract detected in rat plasma were danshensu, protocatechuic acid, caffeic acid, rosmarinic acid, salvianolic acid A, salvianolic acid B, cichoric acid, sinensetin and eupatorin. After liquid-liquid extraction with ethyl acetate, the plasma samples were subjected to a triple quadrupole mass spectrometer employing electrospray ionization (ESI) technique and operating in multiple reaction monitoring (MRM) with both positive and negative ion modes. The standard curves showed good linear regression (r > 0.9915) over the concentration range for the nine analytes. The inter-day and intra-day precision and accuracy were found to be within 15% of the nominal concentration. The recovery and stability of nine compounds were all demonstrated to be within acceptable limits. The approach was successfully applied to investigate the pharmacokinetic analysis of the nine bioactive components after oral administration of OS extract in rats.

Qualitative and quantitative analysis on the chemical constituents in Orthosiphon stamineus Benth. using ultra high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

A sensitive and efficient method was established and validated for qualitative and quantitative analysis on the chemical constituents in Orthosiphon stamineus Benth. (O. stamineus) using ultra high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. Based on the retention time and MS spectra, 61 compounds were detected by using ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry. 52 chemical structures in the O. stamineus extracts including 26 phenolic acids, 11 flavonoids, 6 diterpenoids, 4 fatty acids and 5 tanshinones were tentatively identified without the time-consuming process of isolation. Moreover, five chemical constituents (Danshensu, Caffeic acid, Rosmarinic acid, Sinensetin and Eupatorin) were quantified in three different batches of O. stamineus samples by the developed ultra high-performance liquid chromatography coupled with electrospray ionization triple-quadrupole mass spectrometry method in 10 min. The method validation of the five compounds was performed with acceptable linearity (R2, 0.9930-0.9997), precision (RSD, 1.87-10.36%), repeatability (RSD, 0.59-4.87%) and recovery (105.30-110.53%, RSD ≤ 13.90%).