LC-QTOF MS Research Articles

Age Association Analysis between Tricarboxylic Acid Metabolites and Neurocognitive Impairment in Persons Living with HIV

ObjectiveInflammation may contribute to neurocognitive impairment (NCI) in people with HIV (PWH). Lipopolysaccharide‐mediated macrophage activation induces a shift from oxidative phosphorylation to glycolysis in‐vitro with accumulation of the tricarboxylic acid (TCA) metabolites succinate (SA) and citrate (CA). These metabolites engage diverse cellular inflammatory pathways and may indicate mitochondrial dysfunction. In this study, we developed and optimized an LC‐MS metabolomic method to allow the simultaneous detection and analysis of plasma SA and CA metabolites in in people with HIV.MethodsGeneralized estimating equations were used in a random sample of older PWH from the prospective, multicenter HAILO cohort. Plasma sample was mixed with methanol‐acetonitrile‐water (5:3:1) and 0.1mg/mL internal standard (ISTD, D4‐citric acid). The mixture was vortexed for 30 minutes and centrifuged at 10,000g for 10 minutes. The supernatants were dried, reconstituted in 0.2% formic acid/0.1mg/ml ISTD. Samples were analyzed using Kinetex ‐ C18 column coupled to an Agilent 6540 Q‐TOF LC‐MS. Metabolites identification was achieved based on retention time and product ions. Calibration curves were obtained by using SA and CA authentic standards spiked into 0.2% formic acid/0.1mg/mL ISTD. Using Agilent MassHunter Qualitative B.07 Software, the ratio of analyte and ISTD peak area was plotted against the corresponding concentration to obtain calibration curves. The peak area and the concentration of each analyte were determined using the peak area ratio. Neurocognition was assessed every 48 wks, where NCI was defined as ≥1 z‐scores ≥2 SD below 0 or ≥2 z‐scores ≥1 SD below 0 on Trailmaking A and B and the Wechsler Adult Intelligence Scale‐Revised Digit Symbol tests.Results200 participants were followed a mean ±SD of 160 ±51 wks with a mean of 4 ±1.1 assessments. The mean age was 51 ±7 yrs, 84% were men, 49% were white non‐Hispanic and 59% were current or past smokers. HIV‐1 RNA levels were <400 copies/ml in 97% and the mean nadir and baseline CD4 counts were 234 and 664 cells/mm3, respectively. The mean education was 14 ±4 yrs and NCI was detected at entry in 53 participants (27%). Age modified associations between each metabolite and NCI (P=0.01 for interactions by age with each metabolite). A 1 SD increase of SA or CA levels associated with a 2.2 [95%CI 1.1, 4.3] or a 1.9 [1.3, 2.8] greater odds of NCI, respectively, in the oldest age‐tertile but not in younger age‐groups (Table).ConclusionAssociations between plasma TCA metabolite levels and NCI were evident among older PWH, possibly implicating inflammatory effects on neurocognition that associate with these metabolites or mitochondrial dysfunction and warrant further study.Support or Funding InformationNHLBI (K23HL116209) and NIAD (P30 AI036219)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Comprehensive investigation of pesticides in Brazilian surface water by high resolution mass spectrometry screening and gas chromatography–mass spectrometry quantitative analysis

In this work, a comprehensive investigation on the occurrence of pesticides in the Paraná 3 hydrographic basin of Paraná State, Brazil, was made by application of wide-scope screening based on ultra-high performance liquid chromatography (LC) and gas chromatography (GC) both coupled to quadrupole time-of-flight mass spectrometry (QTOF MS). The use of two complementary techniques, such as GC-QTOF MS and LC-QTOF MS, allowed screening a large number of compounds with different polarity and volatility. This screening approach was applied to 17 samples, enabling the detection of fifty-two pesticides and six metabolites. In a second step, an specific research was made on the herbicide atrazine, one of the most frequent compounds in samples, and its major transformation products (TPs), which were quantitatively analyzed by dispersive liquid-liquid microextraction (DLLME) followed by GC–MS measurement. Twenty-one agricultural streams from the Paraná 3 hydrographic basin were sampled twice in 2017, each time along six successive weeks. Additional samples were also collected after rain events exceeding 10 mm. In total, 407 samples were quantitatively analyzed by DLLME/GC–MS. Atrazine concentrations did not exceed the maximum permitted concentration of 2 μg L−1 according to Brazilian legislation, and only one surface water sample, collected after precipitation events, was slightly above this value (2.89 μg L−1). The maximum concentrations for the TPs desethylatrazine and deisopropylatrazine were 0.80 and 1.22 μg L−1, respectively. Based on the quantification results, a map was produced showing the occurrence of atrazine and its TPs in the area under study. This is the first time that the presence of agrochemicals is evaluated in the Paraná 3 hydrographic basin.

A special case of medicine in disguise: Tattoo inks containing anaesthetics

This paper describes a case of medicine in disguise: seized tattoo inks containing lidocaine and tetracaine at high concentration. Identification of anaesthetics was performed by LC MS Q-TOF with ESI+ source, by accurate mass measurement and by comparing the fragmentation patterns of molecular ions, at 30 V and 10 V of collision-offset voltage, with reference standards. Quantification was also performed by LC MS Q-TOF on the chromatographic peaks in the extracted ion chromatograms, by calibration curves obtained at different standard concentrations and by standard additions approach. The measurement uncertainty was estimated from validation data. The paper gives also chromatographic parameters, MS and MS/MS data and a quantitation method, with a full validation, of other six “caines”. Thus the paper intends to provide a tool for identification and quantitation of the most common local anaesthetics that could be fraudulently added to tattoo inks. The results here reported show that the seized samples of inks represent a serious health risk owing to the high anaesthetic content – therapeutic-like dosage – found.

Solar-light-active mesoporous Cr-TiO2 for photodegradation of spent wash: an in-depth study using QTOF LC-MS.

A dark-coloured effluent called “spent wash” is generated as an unwanted product in sugarcane-based alcohol distilleries. Most distilleries discharge this effluent into soil or water without any treatment, causing water and soil pollution. Herein, we report chromium-doped TiO2 (Cr–TiO2) as a photocatalyst for the degradation of spent wash colour under natural sunlight. Cr-doped TiO2 nanoparticles were prepared using an aqueous titanium peroxide-based sol–gel method with titanium isopropoxide as the Ti precursor and chromium nitrate as the Cr precursor. To observe the effect of dopant on sol–gel behaviour and physicochemical properties, the Cr concentration was varied in the range 0.5–5 wt%. The crystallization temperature and time were optimized to obtain the required phase of Cr–TiO2. The physicochemical characteristics of the Cr-doped TiO2 catalyst were determined using X-ray diffraction, FE-SEM, FETEM, TG, XPS, the Brunauer–Emmett–Teller (BET) method, FT-IR, Raman, PL, ICP-MS, and UV visible spectroscopy. A shift in the absorption edge of TiO2 by doping with chromium suggested an increase in visible light absorption due to a decrease in the effective band gap. The application potential of the Cr–TiO2 catalyst was studied in the degradation of sugar-based alcohol distillery waste under natural sunlight, and the results were compared with those of undoped TiO2 and Degussa P25 TiO2. Degradation of the spent wash solution was monitored using UV-visible, gel permeation chromatography (GPC), and QTOF LC-MS. GPC and LC-MS showed significant changes in the molecular weight of spent wash colour-forming compounds due to the degradation reaction. QTOF LC-MS analysis suggested that acids, alcohols, glucosides, ketones, lipids, peptides, and metabolites were oxidized to low-molecular-weight counterparts. From the results, 5% Cr–TiO2 showed the highest degradation rate among all Cr–TiO2 samples, undoped TiO2, and Degussa P25 TiO2 under identical reaction conditions, with nearly 68–70% degradation achieved in 5 h.

Palmitic acid ester of tetrahydrocannabinol (THC) and palmitic acid diester of 11-hydroxy-THC — Unsuccessful search for additional THC metabolites in human body fluids and tissues

Fatty acid conjugates of hydroxy-metabolites of tetrahydrocannabinol (THC) or cannabinol have already been reported as metabolites in rats. In the herein presented investigation, palmitic acid esters of THC and its primary metabolite 11-hydroxy-delta9-tetrahydrocannabinol (11-OH-THC) were synthesized using esterification with palmitic acid chloride. Structural elucidation of the products was conducted using nuclear magnetic resonance spectroscopy (NMR) and liquid chromatography coupled to quadrupole time of flight mass spectrometry (LC–QToF–MS).For the confirmation of a previous cannabis use, body fluids (femoral blood, heart blood, urine, bile) of 27 death cases (all with known cannabis use), including adipose tissue homogenates of six of these cases as well as eleven plasma samples (probably all with regular cannabis use, confirmed by a high 11-nor-9-carboxy-delta9-tetrahydrocannabinol (THC-COOH) concentration (except one sample, >200ng/mL), were tested for THC and its main metabolites 11-OH-THC and THC-COOH using gas chromatography coupled to mass spectrometry (GC–MS).These samples as well as further tissue homogenates of autopsy cases (liver, kidney, brain) were additionally tested for the presence of THC palmitic acid ester or 11-OH-THC palmitic acid diester by means of a liquid chromatographic triple quadrupole mass spectrometric (LC–QQQ–MS) method, in order to evaluate a possible presence of these conjugates in humans.In none of the analyzed samples (in total 196 specimens; plasma (N=11), femoral blood (N=23), heart blood (N=25), urine (N=23), bile (N=27), liver (N=27), kidney (N=27), brain (N=27), adipose tissue (N=6)), palmitic acid esters of THC or 11-OH-THC could be proven. Even if the existence of these esters in human samples cannot be ruled out definitely, suitability as cannabis consumption markers does not seem likely based on our findings.

Efficient extraction of estrogen receptor–active compounds from environmental surface water via a receptor-mimic adsorbent, a hydrophilic PEG-based molecularly imprinted polymer

We report an efficient screening procedure for the selective detection of compounds that are actively bound to estrogen receptor (ER) from environmental water samples using a receptor-mimic adsorbent prepared by a molecularly imprinted polymer (MIP). To mimic the recognition ability of ER, we improved the typical MIP preparation procedure using a hydrophilic matrix with a polyethylene glycol (PEG)-based crosslinker and a hydrophobic monomer to imitate the hydrophobic pocket of ER. An optimized MIP prepared with methacrylic acid as an additional functional monomer and estriol (E3), an analogue of 17β-estradiol (E2), exhibited highly selective adsorption for ER-active compounds such as E2 and E3, with significant suppression of non-specific hydrophobic adsorption. The prepared MIP was then applied to the screening of ER-active compounds in sewage samples. The fraction concentrated by the MIP was evaluated by in vitro bioassay using the yeast two-hybrid (Y2H) method and liquid chromatography–quadrupole time-of-flight mass spectrometry (LC–Q-TOFMS). Compared to an authentic adsorbent, styrene–divinylbenzene (SDB)-based resin, the fraction concentrated by the MIP had 120% ER activity in the Y2H assay, and only 25% peak volume was detected in LC–Q-TOFMS. Furthermore, a few ER-active compounds were identified only from the fraction concentrated by the MIP, although they could not be determined in the fraction concentrated by the SDB-based resin due to ion suppression along with high levels of hydrophobic compounds. These results indicated that the newly developed MIP effectively captured ER-active compounds and while allowing most non–ER-active compounds to pass through.

Evaluation of an early twentieth century Afghan herbalist’s preparations

Mahomet Allum was a flamboyant philanthropist and herbalist who worked in South Australia in the early part of last century, whose herbal therapies generated some controversy at the time. Two of his preparations have survived to the present day, a general tonic and a treatment for liver and kidney dysfunction. Given the frequent use of pharmaceutical drugs in "tonics" at the time, toxicological analysis was undertaken at Forensic Science SA, Adelaide with liquid chromatography/quadrupole-time-of-flight mass-spectrometer (LC-QTOF MS), liquid-chromatography/ diode array detector (LC/UV) and gas chromatography/ nitrogen phosphorous- detector/mass-spectrometer (GC-NPD/MS), to look for common drugs. In addition DNA analysis was also undertaken at Trace and Environmental DNA (TrEnD) Laboratory (Curtin University) to evaluate the types of plant products used to make these remedies. The general tonic contained genera from the Triticeae (wheat) family as well as the Medicago family (includes alfalfa), possibly as fillers. Other genera found included Utrica (nettle) and Passiflora (passion flower). The preparation for liver and kidney disease also contained genera from the Medicago family as well as genera Arctostaphylos (bear berry) which has traditionally been used for the treatment of dysuria and bladder stones. No common drugs were found. Thus it appears that the two treatments prepared by Mahomet Allum contained only herbal substances and not adulterant pharmaceutical agents. The herbals identified provide an insight into herbalist practices in the early twentieth century.

Proteomic and metabolomic study of wax apple (Syzygium samarangense) fruit during ripening process.

Wax apple is one of the underutilized fruits that is considered a good source of fibers, vitamins, minerals as well as antioxidants. In this study, a comparative analysis of the developments of wax fruit ripening at the proteomic and metabolomic level was reported. 2D electrophoresis coupled with MALDI-TOF/TOF was used to compare the proteome profile from three developmental stages named immature, young, and mature fruits. In general, the protein expression profile and the identified proteins function were discussed for their potential roles in fruit physiological development and ripening processes. The metabolomic investigation was also performed on the same samples using quadrupole LC-MS (LC-QTOF/MS). Roles of some of the differentially expressed proteins and metabolites are discussed in relation to wax apple ripening during the development. This is the first study investigating the changes in the proteins and metabolites in wax apple at different developmental stages. The information obtained from this research will be helpful in developing biomarkers for breeders and help the plant researchers to avoid wax apple cultivation problems such as fruit cracking.

Metabolic Profiling of Cultivated Bush Tea (Athrixia phylicoides DC.) in Response to Different Pruning Types

Bush tea (Athrixia phylicoides DC.) is a popular medicinal South African indigenous plant and it has been used for many decades as a health beverage and medicine. The objective of the study was to profile metabolites for assessment of quality of bush tea (A. phylicoides DC.) subjected to different pruning levels. Treatments consisted of untreated control, top-branch pruning, middle pruning, and basal pruning arranged in a randomized complete block design (RCBD) using 10 single trees as replications. The liquid chromatography quadrupole time-of-flight mass spectrometry (LC–QTOF–MS) was carried out to annotate the bush tea metabolites present in bush tea. Orthogonal partial least square-discriminatory analysis (OPLS-DA) from 1H nuclear magnetic resonance (NMR) revealed a separation between the basal, middle, top pruning, and the unpruned bush tea plants. The pruned (top) and unpruned tea plants, exhibited higher levels of metabolites than the basal and middle pruned. Pruning bush tea showed a significant effect on accumulation of secondary metabolites and thus could enhance bush tea quality. The study successfully annotated 28 metabolites (compounds), which elucidated canonical differences in pruning treatment of bush tea, as validated through multivariate analysis. Top pruning (apically pruned) resulted in improved metabolite accumulation than other treatment and can be recommended in bush tea cultivation. Future studies to enhance vegetative enhancement after pruning will be evaluated.

Top-Down Lc–Ms Quantitation of Intact Denatured and Native Monoclonal Antibodies in Biological Samples

The requirements for developing antibody biotherapeutics benefit from understanding the nature and relevant aspects of the entire molecule. The method presented herein employs on-line multidimensional LC-quadrupole time-of-flight (QTOF)-MS for the quantitative determination of an antibody isolated from biological samples while maintaining the intact native biologically active conformation of the antibody. Following method optimization for a model antibody, an incurred biotherapeutic in cynomologus monkey was quantified in its intact top-down native conformation. A partial method validation demonstrated acceptable precision and accuracy although improved sensitivity requires further studies. An on-line multidimensional LC-MS approach presents a proof-of-principle example for quantifying an intact, native antibody isolated from an incurred biological sample via immunoaffinity techniques coupled with top-down QTOF LC-MS bioanalysis.

Phenolic compounds and antioxidant activity of twelve grape cultivars measured by chemical and electrochemical methods

Five red (Rondo, Cabernet carol, Merlot, Pinot noir, Cabernet sauvignon), five white (Solaris, Riesling, Johanniter, Aurora, Saphira), and two pink (Freiminer, Gewurztraminer) grape cultivars were investigated. The phenolic content was determined by liquid chromatography–mass spectrometry quadrupole time-of-flight (LC-MS QTof) and ultra-performance liquid chromatography-photodiode array-fluorescence (UPLC-PDA-FL); and the antioxidant activity by two spectrophotometric methods: oxygen radical absorbance capacity (ORAC) and 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS°+), and by cyclic voltammetry (CV). The aim of this study was to determine profile of phenolic compounds of popular cool climate grape cultivars and their antioxidant potential. The used methods were compared and correlations were established between investigated parameters. Total phenolic content ranged from 1354.6 mg/100 g in dry weight (d.w). for Aurora to 4567.9 mg/100 g d.w. for Rondo. Highest antioxidant activity was observed in red cultivars Rondo, Merlot and Pinot noir (24.4, 23.6 and 21.3 mmolTrolox/100 g d.w., ABTS°+ assay), while in pink and white cultivars results not exceeded 20 mmolTrolox/100 g d.w. The aforementioned affect in red grapes was associated with their higher content of phenolic compounds, and the presence of an additional group, i.e. anthocyanins. Cyclic voltammetry measurements of redox potential of phenolics confirmed the results achieved by traditional methods of evaluating antioxidant activity. Red cultivars had the highest anodic curve area.

Molecular Structure Characterization of Riverine and Coastal Dissolved Organic Matter with Ion Mobility Quadrupole Time-of-Flight LCMS (IM Q-TOF LCMS).

Deciphering molecular structures of dissolved organic matter (DOM) components is key to understanding the formation and transformation of this globally important carbon pool in aquatic environments. Such a task depends on the integrated use of complementary analytical techniques. We characterize the molecular structure of natural DOM using an ion mobility quadrupole time of flight liquid chromatography mass spectrometer (IM Q-TOF LC/MS), which provides multidimensional structural information on DOM molecules. Geometric conformation of DOM molecules is introduced into molecular-level analysis via the ion mobility (IM) in the system, and an actual measurement of isomers is achieved for the first time. Our data show that natural DOM molecules from several south Texas rivers and adjacent coastal waters have smaller geometric conformation compared with standard biomolecules. Furthermore, about 10% of all DOM molecules resolved within the detection limit of IM-MS had at least one but no more than four isomers. With acquired geometric and isomeric information, we established a multidimensional database containing 89 natural DOM compounds. This database provides a foundation to expand further, or compare, with DOM data from different seasons and locations.

Inhibitory effect of trichodermanone C, a sorbicillinoid produced by Trichoderma citrinoviride associated to the green alga Cladophora sp., on nitrite production in LPS-stimulated macrophages

From the green alga Cladophora sp. collected in Italy, the marine fungal strain A12 of Trichoderma citrinoviride was isolated, identified and characterized. LC-MS qTOF analysis was applied to perform a metabolic profile of the fungal culture. Chromatographic techniques and spectroscopic methods were used to isolate and characterize the major secondary metabolites produced by this strain in liquid culture. In particular, four known sorbicillinoids (trichodermanone C, spirosorbicillinol A, vertinolide and sorbicillin) were purified and identified, together with 2-phenylethanol and tyrosol. Moreover, metabolomic analysis allowed to detect small amounts of trichodimerol, rezishanone A, 2′,3′-dihydrosorbicillin and bisvertinol. For the first time a significant inhibitory effect on nitrite levels has been shown for trichodermanone C in lipopolysaccharide-stimulated J774A.1 macrophages.

LC-QTOF based Untargeted Metabolites, Bioactive Constituents and Elemental Analysis Associated with Antioxidant Activity in Ficus racemosa L.

Antioxidant is a new hype in naturopathy and at present it has huge demand in cosmetic, nutritional supplementation, human health and pharmacological industry. It reduces cell damages caused by free-radical which are responsible for various ailments like cancer, diabetes mellitus, atherosclerosis, cataracts and inflammation. Different parts viz. leaves, fruits and bark of Ficus racemosa were successively extracted and studied of total phenol content in which methanol and water gave good quantity of phenol content. Antioxidant activity was determined and IC50 values were found 53 ug/ml and 59 ug/ml in methanolic extract of leaves and bark, respectively. Elemental analysis using ICPMS was carried out from leaves, fruits and barks of Ficus racemosa and results indicated that B, Fe and Zn were found in good quantity in leaves and bark. Water and methanolic extract of all the parts of plant were identified for their metabolites by Q-TOF LCMS. The bioactive constituents showed important health properties for human and animals like anti-cancer, antioxidant activity, anti-inflammatory and anti-diabetic. Therefore, present study suggested that Ficus racemosa leaves and bark have huge potential in terms of commercial use as source of antioxidants and bioactive constituents.

In vitro co-culture models to evaluate acute cytotoxicity of individual and combined mycotoxin exposures on Caco-2, THP-1 and HepaRG human cell lines

Deoxynivalenol (DON) and zearalenone (ZEA) are mycotoxins primarily produced by Fusarium species and commonly co-occur in European grains. Some in vitro studies reported synergistic combined effects on cell viability reduction for these two natural food contaminants. However, most of these studies were carried out on conventional cell culture systems involving only one cell type and thus did not include cell-cell communication that is closer to in vivo conditions. In this context, we developed easy bi- and tri-culture systems using the Caco-2 (intestinal epithelial cells), THP-1 (monocytes) and HepaRG (hepatic cells) human cell lines in a proliferating state. Individual and combined cytotoxic effects of DON and ZEA were then assessed using co-cultures during 48 h. In bi-culture systems, results showed that only the highest tested dose of ZEA (IC30) induced a significant reduction in THP-1 viability with both Caco-2 and HepaRG cells cultured in transwells above. On the contrary, only the highest tested dose of DON (IC30) significantly affected HepaRG cell viability located under the Caco-2 cell monolayer. In addition, the DON + ZEA combination seemed to induce higher cytotoxicity than each toxin alone. Mycotoxin quantification in the abluminal compartment by Q-TOF LC-MS suggested uptake of both mycotoxins by the different cell lines. According to the co-culturing cell type, possible cell-cell interactions were also observed. Finally, in the tri-culture system, no cytotoxic effects were observed, regardless of the treatment. These findings highlighted the importance of the proposed models to better decipher toxicological impacts of mycotoxins on more complex cellular systems.

Biodegradation and Detoxification of Malachite Green Dye Using Novel Enzymes from Bacillus cereus Strain KM201428: Kinetic and Metabolite Analysis

Enzyme based degradation of organic pollutants is a promising detoxifying approach due to the promiscuous nature of the enzyme, efficiency, cost effective and ecofriendly. In the present study, we have carried out detailed decoloration and degradation studies on a model triphenyl methane group of dyes (Malachite Green dye (MG)) using a newly isolated enzyme from Bacillus cereus KM201428 under the static condition. Biodegradation of dyes was monitored by UV-VIS spectrophotometer and the resultant metabolites analyzed by Liquid Chromatography–Hybrid Quadrupole Time of Flight Mass Spectrometry (LC–QToF-MS) and Gas Chromatography/Mass Spectrometry (GC - MS). Metabolite analysis results revealed that enzymatic degradation of MG dye resulted in complete mineralization and benzene ring-removal; the latter known for organic dye toxicity. Kinetic study results revealed that first-order kinetic model was best applicable for describing MG dye decoloration. Michaelise-Menten kinetics, Lineweaver–Burk plot and Eadie-Hofstee plot models were used to establish the kinetic parameters for the dye decoloration. Lineweaver–Burk plot provided the best theoretical correlation of the experimental data with maximum rate (Vmax) of 17.70 mg l-1h-1 and Michaelis constant (Km) of 124 mgl-1. Results provide evidence that crude enzyme from Bacillus cereus strain KM201428 offers an effective, renewable, ecofriendly and affordable biotechnology for treatment of industrial effluents polluted with organic dye.

Isolation, identification, optimization, and metabolite profiling of Streptomyces sparsus VSM-30

Deep sea sediment samples of Bay of Bengal (Visakhapatnam) have been analyzed for actinomycetes as an elite source to screen for the production of bioactive metabolites. The actinomycetes strain VSM-30 has an exciting bioactivity profile and was isolated during our systemic screening of marine actinomycetes. It was identified as Streptomyces sparsus based on morphological, physiological, biochemical, and molecular approaches. Response surface methodology regression analysis was carried out to fit the experimental data of each response by the second-order polynomial. The results have proven right interaction among process variables at optimized values of incubation time at 12days, pH at 8, temperature at 30°C, concentrations of starch at 1%, and tryptone at 1% and the data have been adequately fitted into the second-order polynomial models. Under these conditions, the responses (zones of inhibition) of plant pathogenic fungi Aspergillus niger, Aspergillus flavus, Fusarium oxysporum, Fusarium solani, and Penicillium citrinum were also matched with experimental and predicted results. Chemotypic analysis of ethyl acetate extract of the strain was done using LC-Q-TOF-MS revealed the presence of bioactive compounds including tryptophan dehydrobutyrine diketopiperazine, maculosin, 7-o-demethyl albocycline, albocycline M-2, and 7-o-demethoxy-7-oxo albocycline in a negative ion mode. The ethyl acetate extract of actinobacterium has been subjected to gas chromatography and mass spectroscopy (GC-MS) revealed the presence of diverse compounds such as dotriacontane, tetracosane 11-decyl-, diheptyl phthalate, 1-hexadecanesulfonyl chloride, L-alanyl-L-tryptophan, phthalic acid ethyl pentyl ester, 4-trifluoroacetoxyhexadecane, and 1H-imidazole 4,5-dihydro-2,4-dimethyl. Hence, the ethyl acetate extract of Streptomyces sparsus VSM-30 may have antibacterial, antifungal, and antioxidant activities due to the presence of secondary metabolites in ethyl acetate extract. The study also supports marine sediment samples of Bay of Bengal, a promising marine ecosystem remained to be explored for new bioactive compounds.

Partial Characterisation and Colorimetric Characteristics of<i> Sargassum</i> sp. Colorant on Treated Polyester Fabric with Dendrimer

This study focuses on the characterization of pigment compounds presence in Sargassum sp. extracts. The ground powder of dry Sargassum sp. was macerated in in methanol solution at 60°C for 48 hours in an aluminium foil-wrapped conical flask (to provide the dark environment). The extracted colourant was measured with UV-vis Spectrophotometer and Q-TOF LCMS to determine the compounds presence. The extracted colourant also used to dye pretreated and untreated polyester fabric with poly(amidoamine) dendrimer at different concentrations with and without mordants. The exhaustion dyeing with the simultaneous mordanting procedure was carried at 85°C for 60 minutes. The colorimetric properties of the dyed fabrics were then analysed. The results showed that the pigment compounds presence in Sargassum sp. extracts is fucoxanthin and chlorophyll. The application of dendrimer improved K/S values of the dyed fabric. Thus, Sargassum sp. extract is suitable to be used as natural dyes for textile dyeing.

Workflow development for targeted lipidomic quantification using parallel reaction monitoring on a quadrupole-time of flight mass spectrometry

Advances in high-resolution mass spectrometers with faster scanning capabilities and higher sensitivities have expanded these instruments' functionality beyond traditional data-dependent acquisition in targeted metabolomics. Apart from the traditional multiple reaction monitoring strategy, the parallel reaction monitoring (PRM) strategy is also used for targeted metabolomics quantification. The high resolution and mass accuracy of full-scan (MS1) and tandem mass spectrometry (MS/MS) scan result in sufficient selectivity by monitoring all MS/MS fragment ions for each target precursor and simultaneously providing flexibility in assay method construction and post-acquisition data analysis. In this study, using an orthogonal quadrupole-time of flight liquid chromatography-mass spectrometry system (QTOF LC-MS), we investigated the applicability of a large-scale targeted lipidomic assay using scheduled PRM. This method monitored 222 lipids belonging to 15 lipid species in serum. Robustness, reproducibility, and quantitative performance were assessed using chemical standards and serum samples. Finally, we demonstrated the application of this PRM-based targeted metabolomic workflow to systemic lupus erythematosus, a severe autoimmunological disease. Results showed that 63 lipids belonging to 11 lipid species were significantly changed. In summary, at the first time, a robust targeted lipidomic workflow was established using PRM acquisition strategy on a Q-TOF platform, providing another powerful tool for targeted metabolomic analysis.

Enhanced production of berberine in In vitro regenerated cell of Tinospora cordifolia and its analysis through LCMS QToF

Tinospora cordifolia is a prioritized medicinal plant and having an immense medicinal importance especially in Indian medicinal system. But this plant needs a regeneration protocol for its rapid propagation. An efficient regeneration protocol was developed for T. cordifolia using nodal explants. High frequency of multiple shoot formation was induced when the nodal segments were cultured on MS medium supplemented with BAP (1.0 mg L−1) and 2-iP (0.5 mg L−1). The highest mean number of shoots per nodal explant (7.9 ± 0.45) with highest shoot length (9.3 ± 0.48 cm) and 86% response were achieved on this media and hormonal concentration. The optimum rooting was obtained on ½ strength of MS medium augmented with IBA (0.5 mg L−1) with 8.3 ± 0.46 cm root length and 89% response. Micropropagated plantlets were found to be identical with the mother plant when clonal fidelity of these plantlets were analyzed with inter simple sequence repeat (ISSR) marker. The berberine content was analyzed through LCMS QToF and the highest amount was found in in vitro callus (19.8 µg/gm) followed by stem (9.3 µg/gm) and leaves of field-grown plants (8.4 µg/gm). Further, presence of berberine was confirmed by ESI–MS spectra with protonated molecular ions ([M + H]+) at m/z 336. Furthermore, MS–MS fragmentation pattern confirmed for the presence of berberine in both the samples. Both the spectra (standard and samples) showed common peaks for berberine in the form of protonated molecular ions ([M + H]+) at m/z 320, m/z 304, m/z 292, m/z 278 in MS/MS mode. The study revealed that developed protocol is potent for rapid mass propagation of this plant species with high accumulation of important secondary metabolite berberine.