LC-QTOF MS Research Articles

Antimicrobial, Cytotoxic, Antiviral Effects, and Spectroscopic Characterization of Metabolites Produced by Fusarium oxysporum YP9B

The goal of the work is to determine the bioactive pharmaceutical metabolites produced by the Fusarium oxysporum YP9B isolate. Ten new natural compounds were isolated from the ethyl acetate extract of the F. oxysporum YP9B strain. Their structures were elucidated by spectroscopic methods using 1D and 2D NMR, UV, FT-IR, and mass spectra (LC-QTOF MS and GC-FID/MS). Identified compounds were named as; (1-benzyl-2-methoxy-2-oxoethyl)-2-hydroxy-3-methylbutanoate (1), 2-oxo-8-azatricyclo[9.3.1.13,7]-hexadeca-1(15),3(16),4,6,11,13-hexaen-10-one (2), 2,3-dihydroxypropanoic, hexadecanoic anhydride (3a), 2,3-dihydroxypropanoic (9Z)-octadecenoic anhydride (3b), 2,3-dihydroxy-propanoic (9Z,12Z)-octadecadienoic anhydride (3c), 2,3-dihydroxypropanoic (11Z)-octadecenoic anhydride (4a), 2,3-dihydroxypropanoic, (9E,12E)-octadecadienoic anhydride (4b), 3-hydroxy-1,2,6,10-tetramethylundecyl hexzadecanoate (5a), 3-hydroxy-1,2,6,10-tetramethylundecyl (9E)-octadecaenoate (5b), and 3-hydroxy-1,2,6,10-tetramethylundecyl octadecanoate (5c). Antimicrobial activities of the isolates obtained from the YP9B strain were determined. Cytotoxic and antiviral activities were tested for the isolates against VERO, MCF-7, PC-3, and A549. Compounds 5a-c, 1, and 3a-c showed bacteriostatic activity at low concentrations, and 4a-b and 2 were found to be bactericides. MIC and MBC values against Mycobacterium smegmatis for the compounds 5a-c and 1 were determined to be ​​ <0.5 µg/mL and 0.46 µg/mL, respectively. The experimental result showed that compounds 2, 5a-c and 1 have strong cytotoxic (7.51±1.38 and 19.13± 0.68 (µM) IC50) activity. The antiviral activity against HSV type-1 was determined to be 1.25 µM for compounds 4a-c and 0.312 µM for compound 1.

Long-Term Stability of 13 Fentanyl Analogs in Blood.

Fentanyl analogs continue to play a major role in proliferating the opioid epidemic in the USA. With high rates of overdose deaths, forensic laboratories experience backlogs, which may lead to false-negative results due to drug instability. To address this issue, a quantitative method was validated for fentanyl analogs (3-methylfentanyl, 4-anilino-N-phenethylpiperidine (4-ANPP), 4-fluoro-isobutyrylfentanyl (4-FIBF), acetylfentanyl, acrylfentanyl, butyrylfentanyl, carfentanil, cyclopropylfentanyl, fentanyl, furanylfentanyl, methoxyacetylfentanyl, p-fluorofentanyl and valerylfentanyl) in blood using liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS) and used to assess long-term stability under various temperature conditions (-20°C, 4°C, ∼25°C and 35°C) for 9 months. Authentic specimens were also analyzed 6 months apart for applicability to postmortem blood. Method validation resulted in calibration ranges of 1-100 ng/mL and limits of detection of 0.5 ng/mL. Precision and bias were acceptable (within ±7.2% coefficient of variation (CV) and ±15.2%, respectively). Matrix effects exhibited ion enhancement for all analytes, except carfentanil and 4-ANPP in low-quality control (>25%). For long-term stability, fentanyl analogs (except acrylfentanyl) remained stable under room temperature and refrigerated conditions at low and high concentrations (81.3-112.5% target) for 9 months. While most fentanyl analogs remained stable frozen, degradation was observed after 2 weeks (four freeze/thaw cycles). At elevated temperatures, most analytes were stable for 1 week (74.2-112.6% target). Acrylfentanyl was unstable after 24 h under elevated (70% loss) and room temperatures (53-60% loss), 48-72 h when refrigerated (28-40% loss) and 4 weeks when frozen (22% loss). In authentic bloods (n = 7), initial furanylfentanyl (FuF) and 4-ANPP concentrations were 1.1-3.6 and 1.4-6.4 ng/mL, respectively. Percentage loss of FuF and 4-ANPP over 6 months were 16.3-37.4% and 0.2-26.8%, respectively. Samples suspected to contain fentanyl analogs are recommended to be stored refrigerated or frozen with limited freeze/thaw cycles. Due to instability, in the event of an acrylfentanyl overdose, samples should be analyzed immediately or stored frozen with analysis within 1 month.

Mycobiota dynamics and mycotoxin detection in PGI Salame Piemonte

The complex mycobiota that colonizes traditional fermented sausages plays an important role in the organoleptic properties of such products. The aim of the present study was to investigate fungal diversity and mycotoxin production during maturation of PGI Salame Piemonte. Casing and meat samples were collected at five sampling times from three different batches produced in the same factory and analysed using culture-dependent and independent approaches. Penicillium nalgiovense, which was deliberately inoculated, and Debaryomyces hansenii were the most dominant taxa in casings. Several other fungi mainly belonging to Penicillium crustosum, Penicillium glabrum, Penicillium nordicum, Cladosporium spp., Candida sake, Candida zeylanoides and Yarrowia divulgata were also identified. The casing mycobiota was compared to that of the meat using a metataxonomic approach and a higher fungal diversity was observed in meat as compared to casings. Mycotoxins and penicillin G were monitored using QTOF LC-MS and only trace amounts of roquefortine C were detected in two batches. The present study highlighted the diversity of Salame Piemonte mycobiota and the important contribution of autochthonous fungi to its diversity. The absence of mycotoxins and penicillin G confirmed the high hygienic quality of the studied product regarding fungal and mycotoxin contamination. For the first time, this study provides insights about Salame Piemonte mycobiota, which together with the bacterial microbiota and Salame Piemonte process specifications, are responsible for the product organoleptic properties.

Peak alignment for herbal fingerprints from liquid chromatography-high resolution mass spectrometry via diffusion model and bi-directional eigenvalues

To align the herbal fingerprints from liquid chromatography-high resolution mass spectrometry (LC-HRMS) more accurately, an algorithm based on sub-window factor analysis (SFA-HRMS) was proposed in this paper. Initially, transforming raw files into equidistant matrices based on region of interest (ROI) searching and fuzzy matching. And Haar continuous wavelet transform (Haar CWT) was used for peak detection in diffusion enhancement signals subsequently. Then bi-directional eigenvalues between two moving sub-windows were determined to obtain the optimum shifts of each candidate peak. That is to say, two types of eigenvalues were calculated for mass spectra (m/z dimension) and diffusion profiles (rt direction) in this critical step, respectively. The last step in SFA-HRMS algorithm, moving the peak zone and keeping the peak shape as much as possible. As a result, three kinds of LC-HRMS fingerprints were aligned by SFA-HRMS algorithm succesfully, including LC- quadrupole time-of-flight mass spectrometry (qTOF MS), ion trap time-of-flight mass spectrometry (IT TOF MS) and Orbitrap mass spectrometry (Orbitrap MS). Comparing with those of XCMS-online, similar behaviors may be observed in the LC-qTOF MS fingerprints from Radix Bupleuri. Even for LC-IT TOF MS signals with burrs, accurate alignments can also be achieved in the fingerprints under two ionization modes. So then twelve sets of Fructus Aurantii samples were classified by hierarchical cluster analysis and functional principal components analysis. The next LC-Orbitrap MS fingerprint from herbal preparation was also discussed. In summary, SFA-HRMS provides new ideas for peak alignment of LC-HRMS fingerprints from complex herbal samples.

Use of ion mobility-high resolution mass spectrometry in metabolomics studies to provide near MS/MS quality data in a single injection.

The use of ion mobility separations (IMSs) in metabolomics approaches has started to be deeply explored in the last years. In this work, the use of liquid chromatography (LC) coupled to IMS-quadrupole time-of-flight mass spectrometry (QTOF MS) has been evaluated in a metabolomics experiments using single injection of the samples. IMS has allowed obtaining cleaner fragmentation spectra, of nearly tandem MS quality, in data-independent acquisition mode. This is much useful in this research area as a second injection, generally applied in LC-QTOF MS workflows to obtain tandem mass spectra, is not necessary, saving time and evading possible compound degradation. As a case study, the smoke produced after combustion of herbal blends used to spray synthetic cannabinoids has been selected as study matrix. The smoke components were trapped in carbon cartridges, desorbed and analyzed by LC-IMS-QTOF MS using different separation mechanisms (reversed phase and HILIC) and acquiring in both positive and negative mode to widen the chemical domain. Partial Least Squares-Discriminant Analysis highlighted several compounds, and ratio between N-Isopropyl-3-(isoquinolinyl)-2-propen-1-amine and quinoline allowed differentiating between tobacco and herbal products. These two compounds were tentatively identified using the cleaner fragmentation spectra from a single injection in the IMS-QTOF MS, with additional confidence obtained by retention time (Rt) and collisional cross section (CCS) prediction using artificial neural networks. Data from this work show that LC-IMS-QTOF is an efficient technique in untargeted metabolomics, avoiding re-injection of the samples for elucidation purposes. In addition, the prediction models for Rt and CCS resulted of help in the elucidation process of potential biomarkers.

Carbon nanotube embedded ultrafiltration membranes for the treatment of rapid granular multimedia prefiltered beauty hair salon and municipal wastewater

Currently, the world is faced with major challenges of water scarcity and water quality, thus the coupling of wastewater treatment technologies for reuse of reclaimed water is a very attractive option. In this study, polyethersulfone (PES) ultrafiltration (UF) nanocomposite membranes containing carboxylic functionalized multiwalled carbon nanotubes (COOH-MWCNTs) were fabricated via the non-solvent induced phase separation (NIPS) method. The membranes were fabricated using polyethylene (PEG) 400 as an additive. Scanning electron microscopy (SEM) examination of the nanocomposite membranes showed that the membranes had porous cross-sectional morphologies and improved porosity with increased PEG 400 content. These membranes were used as an advanced treatment step for the purification of beauty hair salon wastewater. The membrane containing 8.5 wt% PEG achieved 80.84%, 77.51% and 94.51% turbidity removal of the rapid granular multimedia prefiltered beauty hair salon wastewater, municipal effluent and influent wastewater samples, respectively. Poor rejection of most organics in the wastewater samples by the modified membranes was observed. In addition, significant reduction in organic content by the modified membrane in municipal influent sample was confirmed by quadruple time-of-flight liquid chromatography mass spectrometry (QTOF LC-MS) which indicated reduced peaks intensity. The higher the PEG 400 dosage, the larger the pore sizes and the lower the contact angle values and thus, enhanced pure water flux and lower rejection of organics was observed. Energy dispersive X-ray spectroscopy (EDS) of fouled membrane surfaces indicated that metal ions played a role in cake formation. Pressure had no influence on the fouling mechanism of the membrane containing 8.5 wt% PEG and 100% relative wastewater flux restoration was obtained after chemical cleaning of these membranes. These results suggest that an additional advanced water treatment technology may be required for the effective treatment of beauty hair salon wastewater for reuse purposes.

Proteomic profiling of clinical and environmental strains of Pseudomonas aeruginosa

Pseudomonas aeruginosa is a ubiquitous bacterium, which is able to change its physiological characteristics in response to different habitats. Environmental strains are presumably less pathogenic than clinical strains and whether or not the clinical strains originate from the environment or through inter-host transmission remains poorly understood. To minimize the risk of infection, a better understanding of proteomic profiling of P. aeruginosa is necessary for elucidating the correlation between environmental and clinical strains. Based on antimicrobial susceptibility and patterns of virulence, we selected 12 clinical and environmental strains: (i) environmental, (ii) multidrug resistant (MDR) clinical and (iii) susceptible clinical strains. Whole-cell protein was extracted from each strain and subjected to two-dimensional differential gel electrophoresis (2-D DIGE) and liquid chromatography tandem mass spectrometry quadrupole time-of-flight (LC-MS QTOF). All 12 strains were clustered into 3 distinct groups based on their variance in protein expression. A total of 526 matched spots were detected and four differentially expressed protein spots (p < 0.05) were identified and all differential spots were downregulated in MDR strain J3. Upregulation of chitin binding and BON domain proteins was present in the environmental and some MDR strains, whereas the clinical strains exhibited distinct proteomic profiles with increased expression of serine protein kinase and arginine/ornithine transport ATP-binding proteins. Significant difference in expression was observed between susceptible clinical and MDR strains, as well as susceptible clinical and environmental strains. Transition from an environmental saprophyte to a clinical strain could alter its physiological characteristics to further increase its adaptation.

Pepsin generated camel whey protein hydrolysates with potential antihypertensive properties: Identification and molecular docking of antihypertensive peptides

This study reports the angiotensin converting enzyme (ACE) inhibiting potential of peptic camel whey hydrolysates (WHs) produced under different hydrolysis conditions. WHs displayed ACE inhibitory IC50 values in the range of 0.197–1.307 mg/mL. Among 27 hydrolysates generated, four, with the lowest IC50, were subjected to peptide sequencing using LC-MS QTOF and 185 different peptide sequences were identified. Peptides with a high PeptideRanker score (>0.8) were subsequently studied for possible interactions with ACE using in silico analysis. Based on the interaction of peptides with the hot-spots on the enzyme, PAGNFLP, FCCLGPVPP, PAGNFLMNGLMHR, and PAVACCLPPLPCHM were identified as potential ACE inhibitors. Molecular docking was also employed to identify how the shorter peptides interact in the active site of ACE. Furthermore, due to the importance of renin in managing hypertension, peptides from selected WHs with a potential to interact with renin were predicted using in silico analysis. Molecular docking was employed to predict how the identified peptides, PVAAAPVM and LRPFL, could interact with renin and potentially inhibit it. Thus, this study suggests that enzymatic hydrolysis of camel whey proteins could release peptides with potential anti-hypertensive properties.

Removal of pharmaceuticals in hospital wastewater by solar photo-Fenton with Fe3+-EDDS using a pilot raceway pond reactor: Transformation products and in silico toxicity assessment

This study investigated the removal of six representative pharmaceuticals (PHCs), including dipyrone (DIP), diazepam (DZP), fluoxetine (FXT), paracetamol (PCT), propranolol (PPN), and progesterone (PRG) at 500 µg L−1 in raw hospital wastewater (RHWW) treated by solar photo-Fenton with Fe3+-EDDS in a pilot raceway pond reactor (RPR). The effect of the molar ratio (1:1 and 1:2) on the degradation of PHCs and formation of their transformation products (TPs) during the treatments were studied. Additionally, Quantitative Structure-Activity Relationships (Q)SAR in silico toxicity predictions for all TPs identified were evaluated. Experiments were carried out under natural sunlight with a total volume of 10 L of RHWW. The reactant concentrations were PHCs(mix) (500 µg L−1), Fe3+-EDDS (1:2 and 1:1; 15.35 mg L−1 of iron) and H2O2 (230 mg L−1), with all experiments at circumneutral pH. The H2O2 was carefully dosed during the first 30 min of solar photo-Fenton treatment to avoid its continued excess and inefficient use. The main results obtained indicate the degradation of PHCs is highly favored in Fe3+:EDDS (1:2) providing total consumption of H2O2, increased iron stability, degradation percentages above 77% in most PHCs and total degradation of some TPs formed throughout the process at t30W = 57 min. Twenty-one TPs were identified using LC-QTOF MS associated with a purpose-built database for both molar ratio (1:1 and 1:2) of Fe3+-EDDS. To perform the (Q)SAR predictions of the PHCs and TPs identified, the toxicity, biodegradability, mutagenicity, and bioaccumulation were calculated on the basis of in silico tools. Principal component analysis (PCA) was also performed to compare the toxicity of the TPs, allowing the consideration of two principal components which characterized 88% of the predicted results. Most of the TPs did not show toxicity, mutagenicity or bioaccumulation characteristics.

Characterization of Phytochemicals and Chemical Compounds of Patat Leaves (Phrynium capitatum) as Wrapping Materials for Pesor Doclang

Doclang is a typical food of Bogor. One of the compositions is “pesor” wrapped in patat leaves. Apart from being a food wrapper, patat leaves are used as traditional medicine. Research on the chemical content of patat leaves is still limited and needs to be further developed. In this study, phytochemical testing and chemical content of the extracts of n-hexane, ethyl acetate, and methanol from patat leaves were carried out. Phytochemical tests include alkaloid tests; steroid and terpenoid testing; flavonoid test; saponin test; phenolic hydroquinone; tannin and polyphenol test. Based on phytochemical test results, the extract of patat leaves was known to contain alkaloids, steroids, triterpenoids, flavonoids, saponins, phenolic hydroquinones, and tannins with different distributions at each level of solvent polarity. The instruments used to identify the chemical content of patat leaf extract were GCMS and LCMS QTOF. Through GCMS analysis on n-hexane extract, 114 compounds were identified with a similarity level of ≥ 80% to the library. Among these compounds, confirmed to be terpenoid and steroid compounds. In the LCMS analysis of ethyl acetate extract obtained 25 compounds and 31 compounds for methanol extract. It is suspected that these compounds come from the alkaloid, steroid, flavonoid, saponin, phenolic hydroquinone, or tannin class.

Degradation of pharmaceuticals in wastewater matrices through solar light-driven photocatalyst prepared from petrochemical waste.

Pharmaceuticals, such as dipyrone (DIP), paracetamol (PCT), and propranolol (PPN), are widely used analgesics and beta-blockers with the greatest presence in wastewaters and, consequently, in natural waters. The present work evaluated solar light-driven photocatalyst from petrochemical industrial waste (PW) as a strategy for the degradation of three pharmaceuticals in different water matrices (distilled water-DW, simulated wastewater-SWW, and real hospital wastewater-RHWW). All experiments were carried out in a solar photo-reactor with a capacity of 1 L and the experimental condition employed was a catalyst concentration of 350 mg L-1 at pH 5.0; these conditions were selected considering the Doehlert design validation spreadsheet and the desirability function. All materials prepared were conveniently characterized by zeta potential, small-angle X-ray scattering (SAXS), diffuse reflectance ultraviolet-visible (DRUV), and infrared spectroscopy. According to the results of the characterization, significant differences have been observed between the PW and the photocatalyst such as vibrational modes, optical absorption gap, and acid-basic characteristics on the surface, which suggests the potential use of the photocatalyst in the degradation of contaminants of emerging concern. Based on pharmaceutical degradation, DIP showed the highest photosensitivity (87.5%), and therefore the highest photocatalytic degradation followed by PPN; both compounds achieved final concentrations below the limit of quantification of the chromatographic method in DW. However, PCT was the most recalcitrant pharmaceutical in all matrices. Radicals from chromophoric natural organic matter (NOM) could improve PCT degradation in the SWW matrix (56%). Nevertheless, the results in RHWW showed a matrix effect with decreased the oxidation percentages (DIP-99%; PPN-71%; PCT-17%); hence, the addition of an oxidant such as H2O2 was studied as a pharmaceutical oxidation boost in RHWW. PPN was the molecule most sensitive to this strategy of oxidation (98%). Furthermore, 20 transformation products (TPs) generated throughout the treatment were identified by LC-QTOF MS using a customized TPs database. According to quantitative structure activity relationship (Q)SAR analysis, more than 75% of the TPs identified were not biodegradable. About 35% of them have oral toxicity characteristics indicated by Cramer's rules, and the DIP TPs represent high toxicity for different trophic levels.

Simple and Rapid Method for Determination of Abemaciclib in Human Serum using Supported Liquid Extraction Pretreatment and LC-MS/MS Analysis

We developed a simple and rapid method for the determination of abemaciclib in human serum using supported liquid extraction (SLE) method for pretreatment and LC-MS/MS. Abemaciclib was extracted using SLE method with methyltert-butyl ether (MTBE) as elution buffer, and analyzed by LC-QTOF MS system, LCMS9030 (Shimadzu). Abemaciclib and fluconazole (internal standard) were detected with ESI spray in positive ionization mode, and the transition were set at 507.3/393.1629 for abemaciclib and 307.1/220.0677 for fluconazole. The retention times of abemaciclib and fluconazole were 2.76 and 2.98 min, respectively, and good linearity was obtained from 20–1,000 ng/mL for abemaciclib. The regression equation (weight = 1/x2) describing the calibration curve in human serum was y = 0.0196 x – 0.056 (R2 = 0.999), where y is the peak area ratio of abemaciclib against the IS and x is the nominal concentration of abemaciclib. The intra- and inter-assay accuracy varied between -4.3–1.7%, and the precision varied between 0.90–6.19%. The mean recovery rate of abemaciclib was 87.7 ± 4.3%, and the mean matrix factor was 1.00 ± 0.083. Our method offers speed and simplicity of sample preparation, which is one of great advantages in the analysis of clinical specimens. We believe that the present method will contribute to establishing a methodology for determining the optimal dose of abemaciclib for individual patients. Key words: abemaciclib, SLE, supported liquid extraction, quantification

Tube-in-tube membrane photoreactor as a new technology to boost sulfate radical advanced oxidation processes

This work proposes a tube-in-tube membrane photoreactor, operated in a continuous-mode, to boost the efficiency of peroxydisulfate (PDS), through the photolytic (UV-C radiation) and photocatalytic (TiO2-P25) processes. This new technology can efficiently facilitate the transportation of PDS to the catalyst surface and water to be treated. The ultrafiltration tubular ceramic membrane was used as support for the TiO2-P25 and oxidant-catalyst/water contactor. Tests were performed using a synthetic solution and a municipal secondary effluent, both spiked with a pharmaceutical mix solution (paracetamol (PCT), furosemide (FRS), nimesulide (NMD), and diazepam (DZP); 200 μg L−1 of each). At steady-state regime, the UVC/S2O82−/TiO2 system, with radial PDS addition, showed the highest removal of pharmaceuticals in both matrices. Furthermore, twenty-two transformation products (TPs) were identified by applying LC-QTOF MS technique. Hence, the transformation pathways including hydroxylation in aromatic moiety by an electrophilic attack, electron transfer reactions, cleavage of C−O, C−N bond, H−abstraction and ring opening were proposed. TPs chemical structures were evaluated by in silico (Q)SAR approach using TOXTREE and EPI Suite™ software.

The Application of Trichoderma Strains or Metabolites Alters the Olive Leaf Metabolome and the Expression of Defense-Related Genes

Biocontrol fungal strains of the genus Trichoderma can antagonize numerous plant pathogens and promote plant growth using different mechanisms of action, including the production of secondary metabolites (SMs). In this work we analyzed the effects of repeated applications of selected Trichoderma strains or SMs on young olive trees on the stimulation of plant growth and on the development of olive leaf spot disease caused by Fusicladium oleagineum. In addition, metabolomic analyses and gene expression profiles of olive leaves were carried out by LC–MS Q-TOF and real-time RT-PCR, respectively. A total of 104 phenolic compounds were detected from olive leave extracts and 20 were putatively identified. Targeted and untargeted approaches revealed significant differences in both the number and type of phenolic compounds accumulated in olive leaves after Trichoderma applications, as compared to water-treated plants. Different secoiridoids were less abundant in treated plants than in controls, while the accumulation of flavonoids (including luteolin and apigenin derivatives) increased following the application of specific Trichoderma strain. The induction of defense-related genes, and of genes involved in the synthesis of the secoiridoid oleuropein, was also analyzed and revealed a significant variation of gene expression according to the strain or metabolite applied.

Precision-characterization and quantitative determination of main compounds in Si-Ni-San with UHPLC-MS/MS based targeted-profiling method

Mass spectrometry based precision identification of natural products requires the validation of the reference compounds. This study attempted to develop a LC-QTOF MS combined with LC-TQ MS method to precisely characterize the chemicals of Si-Ni-San (SNS), a classic traditional Chinese medicine formula, which is composed of four medicinal plants, and widely used for the treatments of liver disorders. 74 compounds in SNS were provisionally identified by acquiring MS spectra and MS/MS spectra of the possible chemical features, as well as retrieving small-molecule database. By comparing with the accurate MS/MS spectra of reference compounds, 37 compounds in SNS were precisely identified for the first time. In addition, our effort also successfully assigned the origin of each identified compounds against four medicinal plants. Furthermore, we developed a UHPLC-TQ MS based quantitative-profiling method for simultaneous determination of 37 targeted compounds in the different extracts of the raw SNS and commercial lyophilized powders, enabling to facilitate overall quality control of SNS and associated commercial products. Collectively, our finding precisely characterized the main chemicals in SNS, which also provides a new strategy with LC-MS/MS based chemical profiling to precisely identify a diversity of chemicals in Chinese medicinal plants and associated formulae.

Ganoderma spore powder contains little triterpenoids

BackgroundGanoderma spore is a minuscule germ cell ejected from Ganoderma gills during its growth maturity period, it has been considered with high exploitable potential in health-care products manufacture.MethodsAfter testing sporoderm-broken rate, the triterpenoids in 12 batches of broken and unbroken Ganoderma spore powder (GSP) samples were compared with Ganoderma lucidum fruiting body (GL) by high performance thin-layer chromatography (HPTLC) and further verified by liquid chromatography coupled with electrospray ionization quadrupole-time-of-flight mass spectrometry (LC–QTOF–MS); meanwhile, the dissolution of triterpenoids after bionic extraction was also investigated by HPTLC.ResultsThe sporoderm-broken rate of all the broken GSP samples was over 85%. The relative peak area of triterpenoids in GSP samples were lower than 50% of that in fruiting body, and the dissolution of triterpenoids in artificial gastrointestinal fluid was lower than in methanol.ConclusionsThis study demonstrated that there were little triterpenoids in GSP. Triterpenoids in GSP also seldom be dissolved in artificial gastrointestinal fluid.

Evaluation of Recent Data Processing Strategies on Q-TOF LC/MS Based Untargeted Metabolomics

In this study, some of the recently reported data processing strategies were evaluated and modified based on their capabilities and a brief workflow for data mining was redefined for Q-TOF LC-MS based untargeted metabolomics. Commercial pooled human plasma samples were used for this purpose. An ultrafiltration procedure was applied on sample preparation. Sample set was analyzed through Q-TOF LC/MS. A C18 column (Agilent Zorbax 1.8 μM, 50 × 2.1 mm) was used for chromatographic separation. Raw chromatograms were processed using XCMS - R programming language edition and Isotopologue Parameter Optimization (IPO) was used to optimize XCMS parameters. The raw XCMS table was processed using MS Excel to find reliable and reproducible peaks. Totally 1650 reliable and reproducible potential metabolite peaks were found based on the data processing procedures given in this paper. The redefined dataset was upload into MetaboAnalyst platform and the identified metabolites were matched with 86 metabolic pathways. Thus, two list were obtained and presented in this study as supplement files. The first list is to present the retention times and m/z values of detected metabolite peaks. The second list is the metabolic pathways related with the identified metabolites. The briefly described data processing strategies and dataset presented in this study could be beneficial for the researchers working on untargeted metabolomics for processing their data and validating their results.

Solar photo-Fenton treatment of the anti-cancer drug anastrozole in different aqueous matrices at near-neutral pH: Transformation products identification, pathways proposal, and in silico (Q)SAR risk assessment

Anastrozole (ANZ) is a breast cancer drug that was introduced onto the pharmaceutical market in the 1990s and is still one of the most widely consumed cytotoxic compounds. Due to the persistence of the drug, its continued presence after passing through wastewater treatment plants can lead to harm to aquatic environments. The present study investigates use of the solar photo-Fenton (SPF) process applied for ANZ degradation, considering the fate of ANZ and its transformation products (TPs). The SPF process was performed using different concentrations of ferrous iron (Fe2+) and H2O2 in solutions produced with deionized water (DW) and hospital wastewater (HWW), at pH close to neutrality. When solar irradiation in the SPF process was carried out the best ANZ removal rates were found under the following conditions: (i) for the DW matrix, [ANZ]0 = 50 μg L−1, [Fe2+] = 5 mg L−1, and [H2O2]0 = 25 mg L−1, achieving 95% primary ANZ elimination; (ii) for the HWW matrix, [ANZ]0 = 50 μg L−1, [Fe2+] = 10 mg L−1(multiple additions), and [H2O2]0 = 25 mg L−1, achieving 51% primary ANZ elimination. LC-QTOF MS analysis allowed to identify tentatively five transformation products (TPs) formed during the ANZ degradation process in DW, and two TPs when HWW was used. The main proposed degradation pathways were demethylation and hydroxylation. Different in silico models free available (quantitative) structure-activity relationship ((Q)SAR) software were used to predict the ecotoxicities and environmental fates of ANZ and the TPs. The in silico (Q)SAR predictions indicated that ANZ and the TPs were non-biodegradable compounds. In silico (Q)SAR predictions for mutagenicity and carcinogenicity end-points identified some TPs that require further study. Attention is drawn to the formation of several TPs for which statistical and rule-based positive alerts for mutagenic activities were found, requiring further confirmatory in vitro validation tests.

Myco-valorization approach using entrapped Myrothecium verrucaria ITCC-8447 on synthetic and natural support via column bioreactor for the detoxification and degradation of anthraquinone dyes

Myco-valorization (Myco-Valtn) concept was explored using Myrothecium verrucaria ITCC-8447 via column bioreactor to remove two anthraquinone (ARq) dyes. Among the three myco-valtn approaches free cell dependent enabled more than 80% removal of Anthraquinone violet R (AVR) and Alizarin cyanine green (ACG) after 48 h at 30 ± 2 °C. Enzymatic treatment removed more than 50% AVR and ACG after 10 min. However, synthetic and natural support entrapment on Scotch-Brite® (SB) and Luffa cylindrica (LC) enabled more than 80% removal in SB-ACG and LC-ACG and more than 60% in SB-AVR and LC-AVR in 6th cycle. The regenerated column enhanced the removal of dye and allowed the column to run for multiple cycles. Further after FTIR XRD and LCMS Q-TOF, analysis it was observed that ACG and AVR were completely degraded and laccase played the key roles in the degradation of ACG and AVR and the degraded metabolites of anthraquinone dyes (ACG and AVR) exhibited much less toxicity than the parent dye using phytotoxicity assays.

Comparative Lipidomic Study of Human Milk from Different Lactation Stages and Milk Formulas.

In this report, we present a detailed comparison of the lipid composition of human milk (HM) and formula milk (FM) targeting different lactation stages and infant age range. We studied HM samples collected from 26 Polish mothers from colostrum to 19 months of lactation, along with FM from seven brands available on the Polish market (infant formula, follow-on formula and growing-up formula). Lipid extracts were analysed using liquid chromatography coupled to high-resolution mass spectrometry (LC–Q-TOF–MS). We found that the lipid composition of FM deviates significantly from the HM lipid profile in terms of qualitative and quantitative differences. FM had contrasting lipid profiles mostly across brands and accordingly to the type of fat added but not specific to the target age range. The individual differences were dominant in HM; however, differences according to the lactation stage were also observed, especially between colostrum and HM collected in other lactation stages. Biologically and nutritionally important lipids, such as long-chain polyunsaturated fatty acids (LC-PUFAs) containing lipid species, sphingomyelines or ether analogues of glycerophosphoethanoloamines were detected in HM collected in all studied lactation stages. The observed differences concerned all the major HM lipid classes and highlight the importance of the detailed compositional studies of both HM and FM.