Chromatography-electrospray Ionization-quadrupole Time Research Articles

Evaluating the Root Extract of Reynoutria ciliinervis (Nakai) Moldenke: An Analysis of Active Constituents, Antioxidant Potential, and Investigation of Hepatoprotective Effects in Rats

Reynoutria ciliinervis (Nakai) Moldenke (R. ciliinervis) root, a traditional Chinese medicine, was found to exhibit remarkable pharmacological properties through a series of comprehensive investigations. Our study commenced with a qualitative phytochemical analysis that identified 12 bioactive compounds within the plant. Subsequently, utilizing ultraviolet-visible spectrophotometry, the methanol extract emerged as the optimal solvent extract, which was abundant in diverse classes of compounds such as carbohydrates, phenolics, steroids, alkaloids, phenolic acids, and tannins. In vitro antioxidant assays underscored the exceptional free radical scavenging, metal ion chelation, hydrogen peroxide scavenging, singlet oxygen quenching, and β-carotene bleaching capabilities of the methanol extract, significantly outperforming other solvent extracts. Further ultra high-performance liquid chromatography–electrospray ionization–quadrupole time of flight–mass spectrometry analysis revealed the presence of 45 compounds, predominantly anthraquinones and phenolics, in the methanol extract. The extract demonstrated robust stability under various conditions, including high temperatures, varying pH levels, and simulated gastrointestinal digestion as well as efficacy in inhibiting the oxidation in edible oils. Acute toxicity tests in mice confirmed the safety of the methanol extract and provided a valuable dosage reference for future studies. Importantly, high-dose methanol extract exhibited a significant pre-protective effect against D-galactosamine-induced liver injury in rats, as evidenced by reduced alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transpeptidase, malondialdehyde levels, and elevated catalase and albumin levels. These findings suggest a potential role for the methanol extract of R. ciliinervis root in treating oxidative stress-related disorders, highlighting the plant’s immense medicinal potential. Our research offers a thorough evaluation of the bioactive components, antioxidant properties, stability, and liver-protecting effects of the methanol extract, setting the stage for deeper investigation and potential clinical applications.

Rapid identification of chemical compositions from three species of Siegesbeckiae Herba by ultra-performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry in combination with deoxyribonucleic acid barcoding.

Siegesbeckiae Herba, a traditional Chinese medicine, originates from Siegesbeckia orientalis, S. glabrescens, and S. pubescens in the Pharmacopoeia of the People's Republic of China. However, accurate identification of decoction pieces from the three plants remains a challenge. In this study, 26 batches of Siegesbeckiae Herba were identified by deoxyribonucleic acid barcoding, and their chemical compositions were determined using ultra-performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry. The results showed that the internal transcribed spacer 2 and internal transcribed spacer 1-5.8 S- internal transcribed spacer 2 sequences could distinguish three species. In total, 48 compounds were identified including 12 marker compounds screened for three species using the partial least square discriminant analysis. Among these, two diterpenoids 16-O-malonylkirenol and 15-O-malonylkirenol, and a novel diterpenoid 15,16-di-O-malonylkirenol were isolated and identified. A convenient method for the identification of Siegesbeckiae Herba was established using kirenol and 16-O-acetlydarutoside as control standards by thin-layer chromatography. Unexpectedly, none of the batches of S. orientalis contained kirenol, which did not meet the quality standards of Siegesbeckiae Herba, suggesting that the rationality of kirenol as a quality marker for S. orientalis should be further investigated. The results of this study will contribute to the quality control of Siegesbeckiae Herba.

Alternative microbial-based functional ingredient source for lycopene, beta-carotene, and polyunsaturated fatty acids

The acquisition of carotenoids and polyunsaturated fatty acids (PUFAs) from plants and animals for use as functional ingredients raises concerns regarding productivity and cost; utilization of microorganisms as alternative sources is an option. We proposed to evaluate the production of carotenoids and PUFAs by Rhodopseudomonas faecalis PA2 using different vegetable oils (rice bran oil, palm oil, coconut oil, and soybean oil) as carbon source, different concentrations of yeast extract as nitrogen source at different cultivation time to ensure the best production. Cultivation with soybean oil as source of carbon led to the most significant changes in the fatty acid profile. Compared to the initial condition, the strain cultivated in the optimal conditions (4% soybean oil, 0.35% yeast extract, and 14 days of incubation) showed an increase in μmax, biomass, carotenoid productivity, and microbial lipids by 102.5%, 52.7%, 33.82%, and 34.78%, respectively. The unsaturated fatty acids content was raised with additional types of PUFAs; omega-3 [alpha-linolenic acid and eicosapentaenoic acid] and omega-6 [linoleic acid and eicosatrienoic acid] fatty acids were identified. The results of ultra high-performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry (UHPLC-ESI-QTOF-MS/MS) indicated the molecular formula and mass of bacterial metabolites were identical to those of lycopene and beta-carotene. The untargeted metabolomics revealed functional lipids and several physiologically bioactive compounds. The outcome provides scientific reference regarding carotenoids, PUFAs, and useful metabolites that have not yet been reported in the species Rhodopseudomonas faecalis for further use as a microbial-based functional ingredient.

Isolation, Characterization, and Toxicity Study of Stress Degradation Products of Pranlukast Hydrate.

Pranlukast hydrate (PRN), a cysteinyl leukotriene receptor antagonist (CysLT1), is used to treat bronchial asthma. The objective of this study is to perform the isolation, characterization, and toxicity analysis of stress degradation products of PRN. In high-performance liquid chromatography (HPLC), the separation was achieved using a Phenomenex Gemini C18 (250 × 4.6 mm, 5 μ) column; the ammonium format buffer (50 mM), pH 4, with formic acid: acetonitrile (50:50, v/v) was used as a mobile phase at a flow rate of 1.25 mL/min; and the photodiode array detector was used for detection at 230 nm. The drug was subjected to stress degradation as per ICH Q1A (R2) and ICH Q1B guidelines. The drug was found to be labile in alkaline (62.48% degradation) and photolytic (liquid state) (7.67% degradation) conditions, whereas the drug was found to be stable in acidic, peroxide, photolytic (solid state), and thermal conditions. The characterization of the drug and its degradation products was achieved using liquid chromatography-electrospray ionization-quadrupole time of flight tandem mass spectrometry (LC-ESI-QTOF-MS/MS), and the degradation mechanism was proposed. There were two degradation products observed in alkaline conditions (DP6 and DP9), whereas six novel degradation products were observed in photolytic degradation products (DP1, DP3, DP4, DP5, DP7, and DP10). The developed method was successfully validated as per the ICH Q2 (R1) guideline. The isolation of the alkaline degradation product DP9 was performed using preparative HPLC, and it was found to be 96.8% pure degradation product. The characterizations of the isolated degradation product (DP9) and procured impurity were performed using MS/MS, NMR, and FTIR. The mass of the procured impurity and DP9 were observed to be 404 and 500 Da, respectively. The in vitro cytotoxicity study of the procured impurity and DP9 was conducted using a 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay using an A549 cell line, and they were found to be cytotoxic at concentrations above 62.5 and 250 μg/mL, respectively. Furthermore, an in silico toxicity study was performed to predict the toxicity of all the major characterized degradation products of PRN using admetSAR software version 2.0. DP1, DP2, DP6, and DP10 were found to be hepatotoxic, mutagenic according to the micronucleus test, and aquatic toxic. We can conclude that the drug should be kept away from the direct exposure of light and the toxicity levels of DP1, DP2, DP6, and DP10 should be reduced below 0.1% to avoid their toxic effect.

Electrochemical degradation of doxycycline in a three-dimensional vermiculite/peroxymonosulfate electrode system: Mechanism, kinetics, and degradation pathway

In this study, vermiculite (VER) powder was used as a particle electrode in a three-dimensional (3D) Ti-IrO2/Ta2O5 electrode system for the electrochemical degradation of doxycycline (DOX) combined with peroxymonosulfate (PMS) activation. The effects of operational parameters (such as VER dosage, PMS concentration, initial pH, and current density) on DOX degradation were studied and the optimal parameters were discussed. Electrochemical degradation and electrochemical analysis showed that the addition of VER particles could improve the electrochemical conversion efficiency of the 3D electrochemical system by increasing the oxygen evolution potential and inhibiting the occurrence of oxygen evolution side reactions. Quenching experiments showed that sulfate free radicals are the leading reactive oxygen species in the system. A possible pathway for DOX degradation by the reaction system was proposed based on intermediates detected using liquid chromatography–electrospray ionization–quadrupole–time of flight (LC–ESI–Q–TOF). The toxicity of all the intermediates was predicted using the ECOSAR program. Our study showed that the 3D VER/PMS electrode system was reliable for repairing surface water or groundwater polluted by antibiotics.

Elevated α-Ketoglutaric Acid Concentrations and a Lipid-Balanced Signature Are the Key Factors in Long-Term HIV Control.

Long-term elite controllers (LTECs) are a fascinating small subset of HIV individuals with viral and immunological HIV control in the long term that have been designated as models of an HIV functional cure. However, data on the LTEC phenotype are still scarce, and hence, the metabolomics and lipidomics signatures in the LTEC-extreme phenotype, LTECs with more than 10 years of viral and immunological HIV control, could be pivotal to finding the keys for functional HIV remission. Metabolomics and lipidomics analyses were performed using high-resolution mass spectrometry (ultra-high-performance liquid chromatography–electrospray ionization–quadrupole time of flight [UHPLC-(ESI) qTOF] in plasma samples of 13 patients defined as LTEC-extreme, a group of 20 LTECs that lost viral and/or immunological control during the follow-up study (LTEC-losing) and 9 EC patients with short-term viral and immunological control (less than 5 years; no-LTEC patients). Long-term viral and immunological HIV-1 control was found to be strongly associated with elevated tricarboxylic acid (TCA) cycle function. Interestingly, of the nine metabolites identified in the TCA cycle, α-ketoglutaric acid (p = 0.004), a metabolite implicated in the activation of the mTOR complex, a modulator of HIV latency and regulator of several biological processes, was found to be a key metabolite in the persistent control. On the other hand, a lipidomics panel combining 45 lipid species showed an optimal percentage of separation and an ability to differentiate LTEC-extreme from LTEC-losing, revealing that an elevated lipidomics plasma profile could be a predictive factor for the reignition of viral replication in LTEC individuals.

Metabolomics reveals the role of isopentenyl group in coumarins metabolism

Coumarins are a group of natural compounds commonly found in the families of Rutaceae and Umbelliferae. 7-Isopentenyloxycoumarin (ISC), auraptene (AUR), and umbelliprenin (UM) belong to prenyloxycoumarins (PYCs), which link isopentenyl, geranyl, and farnesyl group at C7 position, respectively. The substituent of 7-ethoxycoumarin (ETC) is the ethyl group. In this study, UPLC-ESI-QTOF-MS (ultra-performance liquid chromatography-electrospray ionization-quadrupole time of flight-MS)-based metabolomics was used to evaluate the in vivo and in vitro metabolism of PYCs. Results showed that ETC produced 10 known metabolites, and ISC was transformed into 17 metabolites in vivo and in vitro, which were undescribed compounds. A total of 35 AUR metabolites, including 34 undescribed metabolites were identified, and 21 metabolites were reported for the first time in UM. The results indicated that hydroxylation and N-acetylcysteine conjugation were the common metabolic reactions for PYCs. The metabolic rates of ETC, ISC, AUR and UM were 26%, 36%, 81%, and 38%, respectively, in human liver microsome, while they were 24%, 40%, 80%, and 37%, respectively, in mouse liver microsomes. In addition, recombinant cytochrome P450s (CYPs) screening showed that CYP1A1, 2C19, 3A4, and 3A5 were the major metabolic enzymes involved in the formation of hydroxylation metabolites. Together, these results suggest that the isopentenyl group plays an important role in the metabolism of PYCs.

Proline as a Sparker Metabolite of Oxidative Metabolism during the Flight of the Bumblebee, Bombus impatiens.

Several insect species use the amino acid proline as a major energy substrate. Although initially thought to be limited to blood-feeding dipterans, studies have revealed this capability is more widespread. Recent work with isolated flight muscle showed that the bumblebee Bombus impatiens can oxidize proline at a high rate. However, its role as a metabolic fuel to power flight is unclear. To elucidate the extent to which proline is oxidized to power flight and how its contribution changes during flight, we profiled 14 metabolites central to energy and proline metabolism at key time points in flight muscle and abdominal tissues. Ultra-high performance liquid chromatography-electrospray ionization-quadrupole time of flight mass spectrometry (UPLC-ESI-QTOF MS) analysis revealed that proline is likely used as a sparker metabolite of the tricarboxylic acid cycle at the onset of flight, whereby it supplements the intermediates of the cycle. Carbohydrates are the major energy substrates, which is evidenced by marked decreases in abdominal glycogen stores and a lack of alanine accumulation to replenish flight muscle proline. The time course of fuel stores and metabolites changes during flight highlights homeostatic regulation of energy substrates and patterns of changes in metabolic intermediates within pathways. This study clarifies the role of proline and carbohydrate metabolism during flight in hymenopterans, such as B. impatiens.

Cumyl-CBMICA: A new synthetic cannabinoid receptor agonist containing a cyclobutyl methyl side chain.

Since the beginning of the phenomenon of new psychoactive substances (NPS), synthetic cannabinoid receptor agonists (SCRAs) have been the largest and most prevalent subclass of these drugs in Europe. Many countries implemented specific legislation scheduling classes of substances defined on the basis of their chemical structure to reduce supply. We describe the identification and analytical characterization within the EU project ADEBAR plus of 1-(cyclobutylmethyl)-N-(2-phenylpropan-2-yl)-1H-indole-3-carboxamide which resulted in the formal notification through the Early Warning System of the European Monitoring Centre for Drug and Drug Addiction (EMCDDA). This is the first identification of this new SCRA worldwide and the analytical data was distributed (inter-)nationally right after identification in 2019. First, the substance was isolated from the herbal material using preparative high-performance liquid chromatography (HPLC). Structure elucidation and analytical characterization were performed using gas chromatography-mass spectrometry (GC-MS), gas chromatography-solid state infrared spectroscopy (GC-sIR), liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry (LC-ESI-qToF-MS), Raman spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. The new compound contains a cyclobutyl methyl group as a side chain and has not been described in any patent to our knowledge. Based on the semisystematic nomenclature of SCRAs, we propose Cumyl-CBMICA as a short name for the compound.

Lipidomic Profiling Reveals Significant Perturbations of Intracellular Lipid Homeostasis in Enterovirus-Infected Cells.

Enterovirus A71 (EV-A71) and coxsackievirus A16 (CV-A16) are the most common causes of hand, foot, and mouth disease. Severe EV-A71 and CV-A16 infections may be associated with life-threatening complications. However, the pathogenic mechanisms underlying these severe clinical and pathological features remain incompletely understood. Lipids are known to play critical roles in multiple stages of the virus replication cycle. The specific lipid profile induced upon virus infection is required for optimal virus replication. The perturbations in the host cell lipidomic profiles upon enterovirus infection have not been fully characterized. To this end, we performed ultra-high performance liquid chromatography–electrospray ionization–quadrupole–time of flight-mass spectrometry (UPLC-ESI-Q-TOF-MS)-based lipidomics to characterize the change in host lipidome upon EV-A71 and CV-A16 infections. Our results revealed that 47 lipids within 11 lipid classes were significantly perturbed after EV-A71 and CV-A16 infection. Four polyunsaturated fatty acids (PUFAs), namely, arachidonic acid (AA), docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), and eicosapentaenoic acid (EPA), were consistently upregulated upon EV-A71 and CV-A16 infection. Importantly, exogenously supplying three of these four PUFAs, including AA, DHA, and EPA, in cell cultures significantly reduced EV-A71 and CV-A16 replication. Taken together, our results suggested that enteroviruses might specifically modulate the host lipid pathways for optimal virus replication. Excessive exogenous addition of lipids that disrupted this delicate homeostatic state could prevent efficient viral replication. Precise manipulation of the host lipid profile might be a potential host-targeting antiviral strategy for enterovirus infection.

Lipidomics Profile Changes of Type 2 Diabetes Mellitus with Acute Myocardial Infarction

The morbidity and mortality of cardiovascular disease (CVD)/acute myocardial infarction (AMI) of type 2 diabetes mellitus (T2DM) patients are extremely higher than those without T2DM. Biomarkers can be used to predict the occurrence of acute myocardial infarction, thus effectively reducing the incidence of CVD events, particularly in T2DM patients. Lipids have been shown to be biomarkers and potential therapeutic targets for human diseases. The aim of our study was to investigate the prognostic value of lipid biomarkers for predicting AMI in T2DM patients. A total of 420 subjects were recruited in this research. Liquid Chromatography-Electrospray Ionization-Quadrupole Time of Flight-Mass Spectrometer- (LC-ESI-QTOF-MS-) and Liquid Chromatography/Mass Spectrometer- (LC/MS-) based metabolomic methods were applied to characterize metabolic profiles in each plasma sample. In the first untargeted set, 40 T2DM patients with AMI, 40 T2DM patients without AMI, and 40 control subjects were gender- and age-matched. Eight lipid metabolites showed a significant difference among three groups. Then, in the second set, targeted metabolic profiling assays for these 8 lipid biomarker concentrations in plasma were performed; another 100 T2DM patients with AMI, 100 T2DM patients without AMI, and 100 control subjects were selected independently. Receiver operating characteristic (ROC) curves were constructed, and the area under the ROC curves (AUC) was calculated to determine the potential biomarkers. ROC curve analysis showed that the AUC value of lysophosphatidylcholine (LysoPC) 18:0 is more than 0.7, indicating that LysoPC 18:0 may be a potential sensitive and specific biomarker for T2DM with AMI. The changed plasma concentrations of lipids were significantly associated with T2DM with AMI, which showed great value to be biomarkers, though it requires a prospective cohort study for further validation.

Urolithin Metabotypes Can Determine the Modulation of Gut Microbiota in Healthy Individuals by Tracking Walnuts Consumption over Three Days.

Walnuts are rich in polyphenols ellagitannins, modulate gut microbiota (GM), and exert health benefits after long-term consumption. The metabolism of ellagitannins to urolithins via GM depends on urolithin metabotypes (UM-A, -B, or -0), which have been reported to predict host responsiveness to a polyphenol-rich intervention. This study aims to assess whether UMs were associated with differential GM modulation after short-term walnut consumption. In this study, 27 healthy individuals consumed 33 g of peeled raw walnuts over three days. GM profiling was determined using 16S rRNA illumina sequencing and specific real-time quantitative polymerase chain reactions (qPCRs), as well as microbial activity using short-chain fatty acids analysis in stool samples. UMs stratification of volunteers was assessed using ultra performance liquid chromatography–electro spray ionization–quadrupole time of flight–mass spectrometry (UPLC-ESI-QTOF-MS) analysis of urolithins in urine samples. The gut microbiota associated with UM-B was more sensitive to the walnut intervention. Blautia, Bifidobacterium, and members of the Coriobacteriaceae family, including Gordonibacter, increased exclusively in UM-B subjects, while some members of the Lachnospiraceae family decreased in UM-A individuals. Coprococcus and Collinsella increased in both UMs and higher acetate and propionate production resulted after walnuts intake. Our results show that walnuts consumption after only three days modulates GM in a urolithin metabotype-depending manner and increases the production of short-chain fatty acids (SCFA).

Effect of Thermo-Sonication and Ultra-High Pressure on the Quality and Phenolic Profile of Mango Juice.

Consumer demand for safe and nutritious fruit juices has led to the development of a number of food processing techniques. To compare the effect of two processing technologies, thermo-sonication (TS) and ultra-high pressure (UHP), on the quality of mango juice, fresh mango juice was treated with TS at 25, 45, 65 and 95 °C for 10 min and UHP at 400 MPa for 10 min. The phenolic profile of mango was also analyzed using the newly developed ultra-performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry (UPLC-Q-TOF-HRMSn) and, based on this result, the effect of TS and UHP on the phenolics of mango juice was evaluated. Both treatments had minimal effects on the oBrix, pH, and titratable acidity of mango juice. The residual activities of three enzymes (polyphenol oxidase, peroxidase, and pectin methylesterase), antioxidant compounds (vitamin C, Total phenolics, mangiferin derivatives, gallotannins, and quercetin derivatives) and antioxidant activity sharply decreased with the increase in the temperature of the TS treatment. Nevertheless, the UHP treatment retained antioxidants and antioxidant activity at a high level. The UHP process is likely superior to TS in bioactive compounds and antioxidant activity preservation. Therefore, the mango juice products obtained by ultra-high-pressure processing might be more beneficial to health.

Comprehensive Analysis of Secondary Metabolites in Usnea longissima (Lichenized Ascomycetes, Parmeliaceae) Using UPLC-ESI-QTOF-MS/MS and Pro-Apoptotic Activity of Barbatic Acid

Considering the importance of ultra-performance liquid chromatography-electrospray ionization-quadrupole time of flight-tandem mass spectrometry (UPLC-ESI-QTOF-MS/MS) hyphenated techniques for analysis of secondary metabolites from crude extracts, the present study was aimed at identification of secondary metabolites in acetone extract of the lichen Usnea longissima. From our study, 19 compounds were tentatively identified through comparison of exact molecular masses from their MS/MS spectra, mass fragmentation studies and comparison with literature data. In addition, potent cytotoxic activity of U. longissima extract prompted us to isolate four compounds, 18R-hydroxy-dihydroalloprotolichesterinic acid (19), neuropogolic acid (20), barbatic acid (21), and usnic acid (22) from this extract which were adequately identified through mass spectrometry and NMR spectroscopy. All four compounds displayed cytotoxic activity. Barbatic acid (21) manifested doxorubicin equivalent activity against A549 lung cancer cell line with IC50 of 1.78 µM and strong G0/G1 accumulation of cells. Poly ADP-ribose polymerase (PARP) cleavage confirmed that it induced cytotoxic activity via apoptosis. Finally, our work has discerned the depside, barbatic acid (21) from crude extract as a candidate anti-cancer molecule, which induces cell death by stepping up apoptosis.

Effect of ultrasonic and ball-milling treatment on cell wall, nutrients, and antioxidant capacity of rose (Rosa rugosa) bee pollen, and identification of bioactive components.

Bee pollen has been regarded as a complete nutritional human dietary supplement but its nutrient absorption and biological effects may be restricted by the complex pollen wall. The aim of this study was to explore the effects of ultrasonic and ball-milling treatment on the release of nutritional components and on in vitro and in vivo antioxidant effects of rose (Rosa rugosa) bee pollen. Bee pollen walls were broken to varying degrees, nutrients were released, and in vitro and in vivo antioxidant effects of bee pollen were improved. The scavenging effects of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazolone-6-sulfonic acid) (ABTS) radicals, and oxygen radical absorbance capacity (ORAC) were improved. In aging mice, wall-breaking treatment led to better organ recovery, enhanced superoxide dismutase (SOD) and catalase (CAT) effects, and malondialdehyde (MDA) reduction. Eight compounds of rose bee pollen ethanol extract, including isorhamnetin 3-O-diglucoside and N', N″, N‴-dicaffeoyl p-coumaroyl spermidine were identified by ultra-performance liquid chromatography electrospray ionization quadrupole time of flight mass spectrometry (UPLC-ESI-QTOF-MS/MS) assay. This study showed that ultrasonic treatment had greater wall-disruption effects of bee pollen on nutrient release and antioxidant effect promotion. In conclusion, rose bee pollen, with wall-breaking treatments, may have potential value as an ingredient in functional food processing. © 2019 Society of Chemical Industry.

Diuretic effect and material basis of Clematidis Armandii Caulis in rats

To search for the active diuretic fractions of Clematidis Armandii Caulis( CAC) and determine its main active chemical components by using liquid chromatography-mass spectrometry( LC-MS) and diuretic activity evaluation. CAC 75% ethanol extracts and extracts from different polar solvents were orally administered to saline-loaded rats at different doses. 6 h urinary volume,p H and contents of electrolyte Na+,K+and Cl-were measured. The chemical components of the active fractions were separated and identified by ultra performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry( UPLC-ESI-Q-TOF-MS/MS) method. As compared with the control group,the urine volume was increased by 44%( P< 0. 01) and 34%( P < 0. 05) in CAC75% ethanol extract 57. 74 and 28. 8 mg·kg-1 groups respectively; the Na+excretion was increased by 52%( P< 0. 01) and 45%( P<0. 05),respectively; while the Cl-excretion was increased by 101%( P<0. 01) and 85%( P<0. 05),respectively. The urine volume,Na+excretion and Cl-excretion were increased by 50%( P< 0. 01),58%( P< 0. 05),and 65%( P< 0. 05) respectively in petroleum ether extract 70. 98 mg·kg-1 group as compared with the control group. While for the n-butanol extract 194. 18 mg·kg-1 group,the urine volume,Na+and Cl-excretion were increased by 42%( P<0. 01),41%( P<0. 05) and 97%( P<0. 01),respectively. The diuretic activity of other fractions was not obvious. There was no statistical difference in K+excretion in all groups. The results of LC-MS analysis showed that six compounds,including two sterols,one chromogen and three fatty acids,were identified from petroleum ether extract.Fourteen compounds,including six triterpenoid saponins,six lignin glycosides,one sterol glycoside and one phenolic glycoside,were identified from the n-butanol extract. All the results suggested that the ethanol extract of CAC had remarkable diuretic activity and its main effective components included sterol,triterpenoid saponin and lignin glycosides.

Pharmacokinetics of Sijunzi decoction in rat plasma after oral administration using ultra-high-performance liquid chromatography electrospray ionization quadrupole-time of flight mass spectrometry.

Sijunzi decoction (SJZD) is one of the most well-known traditional Chinese herbal formulations. This study elucidates the pharmacokinetics of SJZD in rat plasma after the administration of a single oral dose of 3 mL/kg using ultra-high-performance liquid chromatography electrospray ionization quadrupole-time of flight mass spectrometry (UHPLC-ESI-Q-TOF/MS) with bergapten as an internal standard. The separation was performed on an Agilent Zorbax Eclipse Plus C18 column (2.1 × 50 mm, 1.8 μm) by elution with acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.3 mL/min. Electrospray ionization in positive and negative ion modes was used to quantify six compounds, with monitored ion m/z values of 249.1397 [M + H]+ and 529.3857 [M + H]+ for atractylenolide III (ATL-III) and pachymic acid (PA), respectively, and m/z of 1107.6638 [M - H]- , 991.5746 [M - H + HCOO]- , 821.3714 [M - H]- , 469.3315 [M - H]- for ginsenoside Rb1, Re, glycyrrhizic acid (GL), and glycyrrhetinic acid (GA), respectively. The calibration curves for ginsenoside Rb1 , Re, ATL-III, PA, GL and GA were 0.0015-0.75, 0.001-0.5, 0.0004-0.2, 0.003-0.9, 0.0015-0.3 and 0.001-1.5 μg/mL, respectively. The intra- and inter-day precisions (RSD) were <14.3%. The rapid, sensitive and specific UHPLC-ESI-Q-TOF/MS method developed and validated in this study was successfully applied to the simultaneous determination of the six components of SJZD using rat plasma for pharmacokinetic studies after oral administration.

Identification of an ACE-Inhibitory Peptide from Walnut Protein and Its Evaluation of the Inhibitory Mechanism.

In the present study, a novel angiotensin I-converting enzyme inhibitory (ACE inhibitory) peptide, EPNGLLLPQY, derived from walnut seed storage protein, fragment residues 80–89, was identified by ultra-high performance liquid chromatography electrospray ionization quadrupole time of flight mass spectrometry (UPLC-ESI-Q-TOF-MS/MS) from walnut protein hydrolysate. The IC50 value of the peptide was 233.178 μM, which was determined by the high performance liquid chromatography method by measuring the amount of hippuric acid (HA) generated from the ACE decomposition substrate (hippuryl-l-histidyl-l-leucine (HHL) to assess the ACE activity. Enzyme inhibitory kinetics of the peptide against ACE were also conducted, by which the inhibitory mechanism of ACE-inhibitory peptide was confirmed. Moreover, molecular docking was simulated by Discovery Studio 2017 R2 software to provide the potential mechanisms underlying the ACE-inhibitory activity of EPNGLLLPQY.

Terpenes and Biological Activities of Euphorbia tortilis

Background: Euphorbia species are known for the diversity of terpenes, and the presence of toxic diterpenoids in Euphorbia species have raised occupational and eco-toxicological concerns. The present study reports the terpene profile of Euphorbia tortilis Rottler ex Ainslie, along with cytotoxicity assay. Method: Triterpenes were isolated from the hexane extract of E. tortilis through column chromatography and were characterized by various spectroscopic techniques. Phorbol esters were detected from the methanol extract of the latex through high performance liquid chromatography-electrospray ionizationquadrupole time of flight tandem mass spectrometry (HPLC-QTOF- MS/MS). Cytotoxicity of the extract was tested against human cervical cancer cell line HeLa and normal cardiac myoblasts cell line H9C2 using MTT assay. Results: The pentacyclic triterpenoids 3β-friedelinol, 3α-friedelinol, friedelin, epi-friedelinyl acetate and taraxerol were isolated and characterized from E. tortilis. Ten phorbol esters belonging to the tigliane family of diterpenoids were detected through HPLC-ESI-QTOF-MS/MS method. The major tumor promoting phorbol ester TPA (phorbol-12-myristate-13-acetate) reported from Euphorbiaceae family could not be detected in E. tortilis, whereas the potential anti HIV and cytotoxic phorbol ester prostratin has been detected. Latex extract was inactive on the normal cell line H9C2 within the concentration range of 10-100μg/ml, while the cell growth inhibition of the extract at 100μg/ml on Hela cell line was negligible (3.06%). Conclusion: Pentacyclic triterpenes were the major compounds isolated and characterized from E. tortilis. HPLC-ESI-QTOF-MS/MS method has been found as an ideal tool for the detection of minor phorbol esters. The terpenoid profile revealed the absence of toxic TPA. Furthermore, the latex extract was non toxic towards normal cell line H9C2. The findings are relevant in the context of toxicological concerns related to Euphorbia members. Keywords: Euphorbia tortilis, triterpene, tigliane diterpenene, phorbol ester, HPLC-ESI-QTOF-MS, cytotoxicity.

Plasma metabolomics reveal the correlation of metabolic pathways and Prakritis of humans

BackgroundAyurveda, an ancient Indian medicinal system, has categorized human body constitutions in three broad constitutional types (prakritis) i.e. Vata, Pitta and Kapha. ObjectivesAnalysis of plasma metabolites and related pathways to classify Prakriti specific dominant marker metabolites and metabolic pathways. Materials and methods38 healthy male individuals were assessed for dominant Prakritis and their fasting blood samples were collected. The processed plasma samples were subjected to rapid resolution liquid chromatography–electrospray ionization–quadrupole time of flight mass spectrometry (RRLC–ESI–QTOFMS). Mass profiles were aligned and subjected to multivariate analysis. ResultsPartial least square discriminant analysis (PLS-DA) model showed 97.87% recognition capability. List of PLS-DA metabolites was subjected to permutative Benjamini–Hochberg false discovery rate (FDR) correction and final list of 76 metabolites with p < 0.05 and fold-change > 2.0 was identified. Pathway analysis using metascape and JEPETTO plugins in Cytoscape revealed that steroidal hormone biosynthesis, amino acid, and arachidonic acid metabolism are major pathways varying with different constitution. Biological Go processes analysis showed that aromatic amino acids, sphingolipids, and pyrimidine nucleotides metabolic processes were dominant in kapha type of body constitution. Fat soluble vitamins, cellular amino acid, and androgen biosynthesis process along with branched chain amino acid and glycerolipid catabolic processes were dominant in pitta type individuals. Vata Prakriti was found to have dominant catecholamine, arachidonic acid and hydrogen peroxide metabolomics processes. ConclusionThe neurotransmission and oxidative stress in vata, BCAA catabolic, androgen, xenobiotics metabolic processes in pitta, and aromatic amino acids, sphingolipid, and pyrimidine metabolic process in kapha Prakriti were the dominant marker pathways.