Quadrupole Ion Research Articles

Investigation of Cytotoxic Constituents from Seed Pulp of Entada Phaseoloides and Metabolite Profiling Using UPLC-QTOF-MSE

Entada phaseoloides (Linn.) Merr. (Family: Fabaceae) is a well-known, traditional, medicinal plant that has been extensively used in the Ayurvedic system of medicine for centuries to combat a wide range of ailments. The goal of this work was to investigate the bioactive constituents from n-butanol extracts of Entada. phaseoloides and develop a method for the comprehensive characterization of saponins using liquid chromatography with an electrospray ionization quadrupole time-of-flight mass spectrometer (LC-ESI-QTOF-MS). A hyphenated technique, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS), has been proposed to integrate LC and MS together with NMR for structural elucidation. This method allowed comprehensive characterization of saponin glycosides from E. phaseoloides based on their MS/MS fragmentation study. The phytochemical study of E. phaseoloides resulted in the isolation and identification of three bio-active constituents. Further, the UPLC-QTOF-MS/MS method led the structure elucidation of saponin constituents directly from crude extracts via comparison of the exact molecular masses from their MS/MS spectra. Identified common fragments m/z 648, 630, 498, 366, and 204 were used for the screening of saponin components. The present study summarizes the isolation and identification of bio-compounds from n-butanol extract and the demonstration of UPLC-QTOF-MS/MS analysis for the characterization of compounds in complex crude extracts. To the best of our knowledge, this is the first systematic study in structural characterization on complex saponins and other metabolites from crude extract of E. phaseoloides using UPLC-ESI-QTOF-MSE. Rapid analysis and characterizations of three new saponins from E. phaseoloides using UPLC-ESI-QTOF-MSE were tentatively identified based on the mass fragmentation study.

Narrow Aperture Detection Electrodes ICR Cell with Quadrupolar Ion Detection for FT-ICR MS at the Cyclotron Frequency.

Ion signal detection at the true (unperturbed) cyclotron frequency instead of the conventional reduced cyclotron frequency has remained a formidable challenge since the inception of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Recently, routine FT-ICR MS at the true cyclotron frequency has become a reality with the implementation of ICR cells with narrow aperture detection electrodes (NADEL). Here, we describe the development and implementation of the next generation of these cells, namely, a 2xNADEL ICR cell, which comprises four flat detect and four ∼45° cylindrical excite electrodes, enabling independent ion excitation and quadrupolar ion detection. The performance of the 2xNADEL ICR cell was evaluated on two commercial FT-ICR MS platforms, 10 T LTQ FT from Thermo Scientific and 9.4 T SolariX XR from Bruker Daltonics. The cells provided accurate mass measurements in the analyses of singly and multiply charged peptides (root-mean-square, RMS, mass error Δm/m of 90 ppb), proteins (Δm/m = 200 ppb), and petroleum fractions (Δm/m < 200 ppb). Due to the reduced influence of measured frequency on the space charge and external (trapping) electric fields, the 2xNADEL ICR cells exhibited stable performance in a wide range of trapping potentials (1-20 V). Similarly, in a 13 h rat brain MALDI imaging experiment, the RMS mass error did not exceed 600 ppb even for low signal-to-noise ratio analyte peaks. Notably, the same set of calibration constants was applicable to Fourier spectra in all pixels, reducing the need for recalibration at the individual pixel level. Overall, these results support further experimental development and fundamentals investigation of this promising technology.

Characterization of the Phenolic Compounds in Different Plant Parts of Amaranthus cruentus Grown under Cultivated Conditions.

Phenolic compounds that are present in amaranth crops have gained a lot of interest from researchers due to their health benefits potential. Therefore, the aim of this study was to investigate phenolic compounds present in different plant parts of Amaranthus cruentus using liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry. Moreover, data were analyzed by one-way analysis of variance of the statistical analysis software, whereas commercial statistical package version 4.02 was used for principal component analysis. A total of 21 phenolic compounds were detected and eight were not identified. Caffeoylsaccharic acid isomer, coumaoryl saccharic acid, tryptophan, feruloyl-d-saccharic acid isomer a, b, and c, caffeoyl isocitrate, quercetin 3-O-rhamnosyl-rhamnosyl-glucoside, feruloyl isocitrate, hyperoside, kaempferol rutinoside, and alkaloid compounds were mostly detected in tender and mature leaves. Generally, rutin content was higher (p < 0.05) in most vegetative parts of the amaranth plant, thus, late maturity leaves, tender leaves, and mature leaves, respectively. Lower quantities of rutin were observed in tender grains, flowers, and mature grains. It can be concluded that amaranth contains phenolic compounds, predominantly in the vegetative parts, which makes it to be a promising source of phenolic compounds beneficial to human health.

The levels of bioactive ingredients in Citrus aurantium L. at different harvest periods and antioxidant effects on H2 O2 -induced RIN-m5F cells.

Citrus aurantium L. (Aurantii fructus) is a multi-purpose citrus fruit with high medicinal and nutritional value, but currently there are no data that can be used to investigate the appropriate harvest time to obtain high-quality citrus bioactive ingredients from it. Phytochemicals and the levels of the main bioactive ingredients were investigated by ultra high performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-Q-TOF/MS). The flavanone, polymethoxyflavone, coumarin, synephrine, and limonin content in the citrus fruit was analyzed at different harvest periods, and significant differences, ranging from 0.03 ± 0.01 to 116.26 ± 40.20 g kg-1 (DW), were shown. These compounds were present in higher amounts in June and then decreased gradually, while the biomass accumulation of most of them showed an increasing tendency around harvest time. The H2 O2 -induced RIN-m5F cells model was employed to evaluate their antioxidant capacity. Citrus fruit harvested from June 11 to July 7 possessed an excellent antioxidant capacity by inhibiting the intensity of intracellular reactive oxygen species (ROS) (P < 0.01) and improving superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH) activity (P < 0.01). The chemical composition and antioxidant capacity of citrus leaves, stems, and roots were also evaluated, and these showed great variation compared with other citrus fruits. Multivariate statistical analysis indicated that harvesting time was related closely to the phytochemical contents and antioxidant capacity. Citrus fruit can be appropriately harvested from June to early July when the levels of bioactive ingredients and antioxidant activity reach higher values. This research provides practical information for producing high-quality citrus products. © 2020 Society of Chemical Industry.

Chemical Profile, Antioxidant, Anti-Inflammatory, and Anti-Cancer Effects of Italian Salvia rosmarinus Spenn. Methanol Leaves Extracts.

In this study, we evaluated and compared the chemical composition, the antioxidant, anti-inflammatory, and anti-proliferative effects of four methanol extracts (R1–R4), of Salvia rosmarinus Spenn. in two different sites of Southern Italy obtained by maceration or ultrasound-assisted extraction. Extracts of S. rosmarinus collected on the Ionian coast are indicated with the abbreviations R1 (maceration) and R2 (ultrasound-assisted extraction). Extracts of S. rosmarinus collected on the Tyrrhenian coast are indicated with the abbreviations R3 (maceration) and R4 (ultrasound-assisted extraction). The chemical composition was analyzed using High Pressure liquid chromatography–Diod-Array detection–Electrospray ionization–Quadrupole–Mass Spectroscopy (HPLC-DAD-ESI-Q-MS). The antioxidant activity was analyzed by 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) 2,2-diphenyl-1-picrylhydrazyl (DPPH), β-carotene bleaching, and Ferric Reducing Antioxidant Power (FRAP) assays. Antioxidant features were also assessed in lipopolysaccharide (LPS)-stimulated RAW-264.7 murine macrophages, evaluating Reactive Oxygen Species (ROS) production; in the same experimental model, the anti-inflammatory activity of the extracts was investigated. Interestingly, all extracts displayed antioxidant and anti-inflammatory properties. They exhibited significative nitrite production inhibitory activity, whith IC50 values ranging from 3.46 to 5.53 µg/mL, without impairing cell viability. The anti-inflammatory activity was also investigated by Western Blotting and immunofluorescence assay, highlighting the R3 and R4 extracts ability to reduce NF-κB translocation, as well as to disrupt the MAPKs signaling pathway. Extracts exhibited both potential anti-proliferative activity on breast cancer cells, inducing apoptosis, without affecting non-tumorigenic cells, and the ability to inhibit MDA-MB-231 cells’ motility. Finally, the rosemary extracts treatment significantly reduced the power of conditioned media, from MCF-7 or MDA-MB-231 cells to induce nitrite production on RAW 264.7 cells, confirming their promising anti-inflammatory activity.

Identification of the absorptive constituents and their metabolites in vivo of Ziziphi Spinosae Semen by UPLC-ESI-Q-TOF-MS/MS.

In this research, ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS/MS) was used for detection and identification of the absorptive constituents and their metabolites in rat plasma, urine and feces following oral administration of Ziziphi Spinosae Semen alcohol extract. After structure elucidation, a total of 12 compounds in rat plasma, comprising seven prototypes and five metabolites, 28 compounds in urine, comprising 17 prototypes and 11 metabolites, and 23 compounds in feces, comrpising 17 prototypes and six metabolites, have been tentatively identified by comparison with standard compounds and reference literature information. To the best of our knowledge, this is the first comprehensive and systematical metabolic study on the seed. Mostly importantly, we propose that gastric acid could convert jujubosides into an absorbable form of ebelin lactone oligosaccharides, which may be responsible for the low bioavailability and specific bioactivities of these compounds. Additionally, we deduced that the absorption site of ebelin lactone oligosaccharides is located in the stomach, and that the ebelin lactone form of jujubosides may be more suitable for absorption than its hydrolysis product. Our investigation will be helpful to narrow the scope for potentially active ingredients of the seed, and pave the way for determination of the pharmacological mechanism of the seed.

Integrated study of metabolomics and gut metabolic activity from ulcerative colitis to colorectal cancer: The combined action of disordered gut microbiota and linoleic acid metabolic pathway might fuel cancer.

Colorectal cancer (CRC) is one of the most serious complications of ulcerative colitis (UC). Altered gut microbiota is implicated in the development of CRC and metabolic perturbations are often associated with changes in the gut microbiome composition. Given the links between gut microbiome and the metabolic profiles in the body, an approach involving ultra-high-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) metabolomics and 16S rDNA sequencing technology was applied to trace the development UC into CRC in rats. The study identified 11 differential metabolites related to both UC and CRC, which mainly referred to the linoleic acid metabolism. Among these, linoleic acid and 12‑hydroxy‑8,10-octadecadienoic acid could serve as key biomarkers for the development of UC into CRC. Besides, a significant change was observed in the microflora structure during the development from UC to CRC; this mainly involved a gradual increase in Escherichia-Shigella and a gradual decrease in Lactobacillus. In addition, Pearson's correlation analysis revealed strong correlations between intestinal microflora-related metabolites and specific intestinal microflora, which indicated both of them can promote the transition of UC to CRC. The results of the present study provided positive support for the involvement of intestinal microflora and host metabolism in the pathophysiological mechanism that is responsible for the development of UC into CRC. This information can help understand the risk for CRC that accompanies a diagnosis of UC and also provide different means of targeting these differential metabolites and intestinal microbiota to avoid UC-induced CRC.

Characterization of Hydrocarbon Groups in Complex Mixtures Using Gas Chromatography with Unit-Mass Resolution Electron Ionization Mass Spectrometry

Complex mixtures of hydrocarbons are ubiquitous as petroleum fuels and, consequently, environmental contaminants. Because they contain thousands of individual components with similar molecular structures, detailed chemical characterization of hydrocarbon mixtures relies on advanced analytical techniques that are not accessible to many researchers. Many analyses of hydrocarbon mixtures instead characterize them as "unresolved complex mixtures", with quantification limited to a small number of resolvable components and/or total observed mass within specified volatility ranges. This work develops a new analytical approach to characterize the hydrocarbon component of petroleum and environmental mixtures by "hydrocarbon group" (defined by carbon number, degree of unsaturation and, in certain cases, degree of branching) using gas chromatography coupled to a unit-mass-resolution electron ionization quadrupole mass spectrometer (GC/EI-MS), a standard and widely available instrument. Average mass spectra of hydrocarbons from a widely used spectral library are combined with chromatographic signal representing the molecular ion of each hydrocarbon group to recreate the magnitude and mass spectra of the chromatogram. Characterization of hydrocarbons in diesel fuel by this approach is in good agreement with state-of-the-art techniques relying on high-resolution and fast-response mass spectrometers. Application of this approach to subsurface soil gas samples from remediated sites of underground storage tank spills demonstrates that composition of hydrocarbons in environmental samples varies significantly and that the total signal of samples from contaminated sites may contain a substantial fraction of oxygenated components.

Optical Fiber-Enabled Photoactivation of Peptides and Proteins.

Photoactivation and photodissociation have long proven to be useful tools in tandem mass spectrometry, but implementation often involves cumbersome and potentially dangerous configurations. Here, we redress this problem by using a fiber-optic cable to couple an infrared (IR) laser to a mass spectrometer for robust, efficient, and safe photoactivation experiments. Transmitting 10.6 μm IR photons through a hollow-core fiber, we show that such fiber-assisted activated ion-electron transfer dissociation (AI-ETD) and IR multiphoton dissociation (IRMPD) experiments can be carried out as effectively as traditional mirror-based implementations. We report on the transmission efficiency of the hollow-core fiber for conducting photoactivation experiments and perform various intact protein and peptide analyses to illustrate the benefits of fiber-assisted AI-ETD, namely, a simplified system for irradiating the two-dimensional linear ion trap volume concurrent with ETD reactions to limit uninformative nondissociative events and thereby amplify sequence coverage. We also describe a calibration scheme for the routine analysis of IR laser alignment and power through the fiber and into the dual cell quadrupolar linear ion trap. In all, these advances allow for a more robust, straightforward, and safe instrumentation platform, permitting implementation of AI-ETD and IRMPD on commercial mass spectrometers and broadening the accessibility of these techniques.

Study on characteristics of biflavanones distribution in Garcinia kola seeds and identification of compounds in gum resin exuded from fresh slices

Garcinia kola seeds play an important role in African traditional medicine, which are widely used to treat stomachache, gastritis, malaria, venereal diseases and laryngitis. The habitats, specification, different parts of seeds and slicing process are important factors that affect the quality of Garcinia kola seeds. Interestingly, brownish yellow or brownish red spots were derived from gum resin exuded during slicing, whose chemical constituents have not been elucidated. A rapid high performance liquid chromatography method was established for quantitative study of four main biflavanones (GB-2, GB-1, kolaflavanone and GB-1a) in Garcinia kola seeds. We found that the content of the tested biflavanones was slightly different in Garcinia kola seeds from different habitats, while the seeds weight showed no obvious influence on the biflavanones content from the same habitat. With the aid of “spider-web” mode, we unveiled that the accumulation of biflavanones mainly occurred in seed kernels. Furthermore, slices thickness (1–2 mm) was optimized by comprehensive evaluation of biflavanones’ content, moisture, cutting-time and drying-time to efficiently dry the slices. By employing of ultra-high performance liquid chromatography coupled with electrospray ionization quadrupole Orbitrap high resolution mass spectrometry (UHPLC/ESI Q-Orbitrap MS), garcinoic acid and its derivatives were successfully identified in gum resin, conducing to the interesting finding about the distribution of biflavanones and derivatives of garcinoic acid in Garcinia kola seeds. Generally, our findings in this research could contribute to standardizing the process of harvesting, processing and quality control for Garcinia kola seeds, and help better development and utilization of this important medicinal plant.

Limonoids From Choerospondias axillaris

Eight limonoids (1-8), including 1 new compound named as axillariol A (1), were isolated from the dichloromethane extract of Choerospondias axillaris branches and leaves using various chromatographic separations. Their structures were confirmed by detailed analysis of the 1-dimensional (1D), 2D nuclear magnetic resonance, and high-resolution electrospray ionization quadrupole time-of-flight mass spectra. In addition, toonaciliatins N (6) and toonaciliatins O (7) showed selective and significant cytotoxicity on the rhabdomyosarcoma human cancer cell line with half-maximal inhibitory concentration values of 9.12 and 7.79 µM, respectively.

Metabolites identification and reversible interconversion of chelerythrine and dihydrochelerythrine in vitro/in vivo in rats using ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry

Chelerythrine (CHE) and dihydrochelerythrine (DHCHE), two typical benzophenanthridine alkaloids, have a wide range of pharmacological activities, such as antibacterial, anti-tumour and antiparasitic activities. To date, the biological activities of CHE and DHCHE are well reported, but the biotransformation of CHE and DHCHE in vivo remains unknown. This study aims to clarify the metabolic pathway of CHE and DHCHE in rat liver microsomes (RLMs) in vitro and in vivo. An ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC/ESI-QTOF-MS) method was developed for metabolites identification of CHE and DHCHE. The urine, feces, bile, and plasma samples and RLMs samples were collected for analyzing the biotransformation pathway of CHE and DHCHE. The result showed that there is a phenomenon of mutual reversible interconversion between CHE and DHCHE in vivo and in vitro. The other biotransformation pathways of CHE and DHCHE including demethylation, hydroxylation, methylene dioxy cycle opening, and glucuronidation mainly occurred in the side chain of benzophenanthridine parent structure. Twenty-five phase I and eight phase II metabolites of CHE, twenty-two phase I and eight phase II metabolites of DHCHE were detected. The results will help to develop a deeper understanding of CHE and DHCHE in vivo process and provide some references for the biotransformation research of other benzophenanthridine alkaloids.

Ultra-high-performance liquid chromatography supports a new reaction mechanism between free radicals and ferulic acid with antimicrobial and antioxidant activities

The chemical reaction involving phenolic antioxidants from plant´ extracts have not been adequately studied. In this study, we investigate a quick, accurate, and innovative strategy to elucidate the ferulic acid reaction employing a conclusive analytical tool (ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry, UHPLC-ESI-Q-TOF-MS/MS) used in pharmaceutical, cosmetic, food, and biomedical applications. The ferulic acid- DPPH(2,2-diphenyl-1-picrylhydrazyl) reaction presented three chromatographic peaks with retention times at 5.95, 8.69, and 9.04 min. The peak at 385.0928 mass/charge (m/z) is assigned to the ferulic acid-ferulic acid dimer, while the m/z 586.1193 signal indicates the adduct presence. This statement is confirmed by the fragmentation step, which supports characteristics peaks ascribed to the original DPPH at m/z 195.9986 and m/z 225.9972. The peak m/z 778.1605 is attributed to the formation of a new compound (called radimerSB) that has never identified before. According to the reaction mechanism, the ferulic acid may provide antioxidant activity mainly by hydrogen-atom transfer reaction. Furthermore, we compare the antimicrobial action of ferulic acid and Trolox. We show that against ferulic acid prevents the crop growth of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). The ferulic acid biological activities can be useful to develop new bio-based materials for technological applications (e.g., medical purposes). It is essential to understand the reaction mechanism between DPPH and antioxidants. The DPPH can quantify the concentration of antioxidants in plant extracts. The biological activities (antimicrobial, cytotoxicity, and others) of these extracts depend on the type and concentration of antioxidants and their reaction products.

Plasma Lipid Profile Reveals Plasmalogens as Potential Biomarkers for Colon Cancer Screening.

In this era of precision medicine, there is an increasingly urgent need for highly sensitive tests for detecting tumors such as colon cancer (CC), a silent disease where the first symptoms may take 10–15 years to appear. Mass spectrometry-based lipidomics is an emerging tool for such clinical diagnosis. We used ultra-performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry operating in high energy collision spectral acquisition mode (MSE) mode (UPLC-QTOF-MSE) and gas chromatography (GC) to investigate differences between the plasmatic lipidic composition of CC patients and control (CTR) subjects. Key enzymes in lipidic metabolism were investigated using immuno-based detection assays. Our partial least squares discriminant analysis (PLS-DA) resulted in a suitable discrimination between CTR and CC plasma samples. Forty-two statistically significant discriminating lipids were putatively identified. Ether lipids showed a prominent presence and accordingly, a decrease in glyceronephosphate O-acyltransferase (GNPAT) enzyme activity was found. A receiver operating characteristic (ROC) curve built for three plasmalogens of phosphatidylserine (PS), named PS(P-36:1), PS(P-38:3) and PS(P-40:5), presented an area under the curve (AUC) of 0.998, and sensitivity and specificity of 100 and 85.7% respectively. These results show significant differences in CC patients’ plasma lipid composition that may be useful in discriminating them from CTR individuals with a special role for plasmalogens.

LC-ESI-QTOF-MS/MS Characterization of Seaweed Phenolics and Their Antioxidant Potential.

Seaweed is an important food widely consumed in Asian countries. Seaweed has a diverse array of bioactive compounds, including dietary fiber, carbohydrate, protein, fatty acid, minerals and polyphenols, which contribute to the health benefits and commercial value of seaweed. Nevertheless, detailed information on polyphenol content in seaweeds is still limited. Therefore, the present work aimed to investigate the phenolic compounds present in eight seaweeds [Chlorophyta (green), Ulva sp., Caulerpa sp. and Codium sp.; Rhodophyta (red), Dasya sp., Grateloupia sp. and Centroceras sp.; Ochrophyta (brown), Ecklonia sp., Sargassum sp.], using liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS). The total phenolic content (TPC), total flavonoid content (TFC) and total tannin content (TTC) were determined. The antioxidant potential of seaweed was assessed using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, a 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) free radical scavenging assay and a ferric reducing antioxidant power (FRAP) assay. Brown seaweed species showed the highest total polyphenol content, which correlated with the highest antioxidant potential. The LC-ESI-QTOF-MS/MS tentatively identified a total of 54 phenolic compounds present in the eight seaweeds. The largest number of phenolic compounds were present in Centroceras sp. followed by Ecklonia sp. and Caulerpa sp. Using high-performance liquid chromatography-photodiode array (HPLC-PDA) quantification, the most abundant phenolic compound was p-hydroxybenzoic acid, present in Ulva sp. at 846.083 ± 0.02 μg/g fresh weight. The results obtained indicate the importance of seaweed as a promising source of polyphenols with antioxidant properties, consistent with the health potential of seaweed in food, pharmaceutical and nutraceutical applications.

Rapid Determination of Furoquinoline Alkaloids in Rutaceae Species by Ultra-Performance Liquid Chromatography (UPLC) with Photodiode Array (PDA) and Electrospray Ionization–Quadrupole Time-of-Flight Mass Spectrometry (ESI-Q-TOF/MS)

A qualitative and quantitative method based on ultra-performance liquid chromatography (UPLC) and photodiode array (PDA) and electrospray ionization–quadrupole time-of-flight mass spectrometry (ESI-Q-TOF/MS) was developed, verified, and used to determine the quinoline alkaloids in Dictamnus dasycarpus and Zanthoxylum schinifolium, belonging to the Rutaceae family. Excellent analyte peak resolution was achieved on both quaternary and binary pumps by using acetonitrile and water containing 0.1% formic acid as the mobile phase using a C18 column. All alkaloids exhibited maximum absorption wavelength from 239 to 244 nm. The analytes were ionized as quasi-molecular ions [M + H]+ in MS and were characterized by using fragmentation ions formed by collision-induced dissociation (CID). This information was included in a library that was used to qualitatively identify alkaloids. The proposed method showed excellent linearity (r2 > 0.9988), limits of detection (PDA ≤ 25 ng/mL, MS ≤ 1.2 ng/mL), limits of quantification (PDA ≤ 75 ng/mL, MS ≤ 3.6 ng/mL), precision (in terms of relative standard deviation) of PDA ≤ 1.81% and MS 5.7%,. Liquid–liquid extraction (LLE) and solid-phase extraction (SPE) were conducted to separate and purify the alkaloids from the extracts. The analyte recoveries were determined to be PDA ≤ 103.11% and MS ≤ 104.45%. The method proceeded to deliver the comparable results for the determination of alkaloid components in samples. The developed UPLC-PDA and UPLC-ESI-Q-TOF/MS protocol facilitated the rapid, qualitative and quantitative identification of alkaloids in plants.

Natural Deep Eutectic Solvent as Extraction Media for the Main Phenolic Compounds from Olive Oil Processing Wastes.

In this new century, sustainable development challenges chemical sciences to develop new and clean technological processes. The agri-food industry produces significant quantities of waste, raising significant economic and environmental concerns. Food waste valorization using environmentally friendly procedures is of increasing importance. This study describes the use of several Natural Deep Eutectic Solvents (NADESs) for the microwave-assisted extraction (MAE) of valuable bioactive phenolic compounds from olive oil processing wastes. The extracted samples were characterized by liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS) analysis and the quantification of the phenolic compounds was performed by HPLC analysis. The obtained data were compared with those obtained using water as the solvent in the same extraction conditions. The extraction process is nontoxic, simple and selective and meets most of the criteria to be considered as a sustainable process, with the solvents arising directly from nature.

Qualitative analysis of enzymatic and chemical depolymerized low molecular weight heparins by UHPLC coupled with electrospray ionization quadrupole time-of-flight-mass spectrometry.

Complete heparin digestion with heparin lyase I and II results in a mixture of hexasaccharides and tetrasaccharides with 3-O-sulfo group-containing glucosamine residues at their reducing ends. Because these tetrasaccharides are derived from antithrombin III-binding sites of heparin, we examined whether this method could be applied to estimate the anticoagulant activity of heparin. Therefore, this paper presents a new low molecular weight heparin sample preparation method-chemical depolymerization. Qualitative analysis of the studied compounds and a comparison of their composition are an important contribution to the structural analysis of low molecular weight heparins, which has not been fully conducted so far. Qualitative on-line liquid chromatography-mass spectrometric analysis of these resistant oligosaccharides is also described in this paper.

Systematic analysis and identification of the absorption and metabolic components of Zengye decoction in type 2 diabetic rats by HPLC-ESI-Q-TOF\u2013MS/MS

BackgroundZengye decoction (ZYD) has been widely used in the treatment of type 2 diabetes mellitus (T2DM). Exploring the fate of various components of ZYD in vivo is of considerable significance for pharmacological research and molecular mechanism elaboration. However, the systematic analysis on the metabolic behavior of chemical components of ZYD in T2DM rats has not been reported.MethodsTo screen and characterize the complex chemical compositions of ZYD, and metabolism fate in plasma, urine, bile, and feces of T2DM rats, the model of T2DM rats was prepared. A rapid procedure using high-performance liquid chromatography coupled with electrospray ionization quadrupole time of flight tandem mass spectrometry (HPLC-ESI-Q-TOF–MS/MS) was established. Data were acquired and analyzed by Agilent MassHunter Workstation Qualitative Analysis software version B.07.00 and PCDL manager B.07.00.ResultsA total of 80 compounds were identified or tentatively characterized in ZYD, 31 more than previously detected. Besides, 36 prototype components and 49 metabolites of ZYD were found and characterized in T2DM rats, and the proposed fragmentation pathways and possible metabolic behaviors of the main types of compounds were described.ConclusionsThis study developed the understanding of the composition of ZYD as well as the cleavage rules and metabolic pathways of the prototype compounds. Besides, this study provided abundant data for further research and for study of the metabolism of traditional Chinese medicine prescriptions.

Black Bean Anthocyanin-Rich Extract from Supercritical and Pressurized Extraction Increased In Vitro Antidiabetic Potential, While Having Similar Storage Stability.

Black bean is a source of anthocyanins and other phenolic compounds that are associated with health benefits. This work aimed to optimize the extraction and determine the stability and biological potential of black bean anthocyanin-rich extracts recovered by supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE). The highest concentration of anthocyanins and total phenolic compounds were recovered with SFE using 300 bar, 60 °C and co-solvent ethanol/distilled water (50/50, v/v). Eleven non-colored phenolic compounds were identified in SFE extract using Ultra performance liquid chromatography - Electrospray ionization–Quadrupole -Time of flight - Mass spectrometry (UPLC-ESI-QToF-MS/MS). Myricetin, syringic acid, rutin hydrate and chlorogenic acid presented the highest relative area among identified compounds. Compared to leaching extraction, SFE extracts showed a similar storage stability at 4, 25 and 32 °C (p < 0.05), but with a higher antioxidant potential (2,2-diphenyl-1-picryl-hydrazil (DPPH) IC50: 0.078 ± 0.01; 2,2’-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) IC50: 0.161 ± 0.03) and antidiabetic potential (α-amylase IC50: 124.76 ± 12.97; α-glucosidase IC50: 31.30 ± 0.84; dipeptidyl peptidase-IV IC50: 0.195 ± 0.01). SFE extraction is an efficient method to obtain anthocyanins and other phenolic compounds with exceptional biological potential.