scholarly journals UHPLC-HRMS-Based Untargeted Lipidomics Reveal Mechanism of Antifungal Activity of Carvacrol against Aspergillus flavus

Foods ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 93
Author(s):  
Chenling Qu ◽  
Zhuozhen Li ◽  
Xiupin Wang
Keyword(s):  
Aspergillus Flavus ◽  
High Performance ◽  
High Resolution Mass Spectrometry ◽  
Toxin Production ◽  
Negative Ion ◽  
Mycelia Growth ◽  
Antifungal Substances ◽  
Negative Ion Mode ◽  
Positive Ion ◽  
Treatment Concentration

Aspergillus flavus is a common contaminant in grain, oil and their products. Its metabolite aflatoxin B1 (AFB1) has been proved to be highly carcinogenic. Therefore, it is of great importance to find possible antifungal substances to inhibit the growth and toxin production of Aspergillus flavus. Carvacrol (CV) was reported as a potent antifungal monoterpene derived from plants. In this paper, the antifungal effects and mechanism of CV on Aspergillus flavus were investigated. CV was shown good inhibition on the growth of Aspergillus flavus and the production of AFB1. CV used in concentrations ranging from 0, 50, 100 and 200 μg/mL inhibited the germination of spores, mycelia growth and AFB1 production dose-dependently. To explore the antifungal mechanism of CV on Aspergillus flavus, we also detected the ergosterol content of Aspergillus flavus mycelia, employed Scanning Electron Microscopy (SEM) to observe mycelia morphology and utilized Ultra-High-Performance Liquid Chromatography-High-Resolution Mass Spectrometry (UHPLC-HRMS) to explore the lipidome profiles of Aspergillus flavus. The results showed that the production of ergosterol of mycelia was reduced as the CV treatment concentration increased. SEM photographs demonstrated a rough surface and a reduction in the thickness of hyphae in Aspergillus flavus treated with CV (200 µg/mL). In positive ion mode, 21 lipids of Aspergillus flavus mycelium were downregulated, and 11 lipids were upregulated after treatment with 200-µg/mL CV. In negative ion mode, nine lipids of Aspergillus flavus mycelium were downregulated, and seven lipids upregulated after treatment with 200-µg/mL CV. In addition, the analysis of different lipid metabolic pathways between the control and 200-µg/mL CV-treated groups demonstrated that glycerophospholipid metabolism was the most enriched pathway related to CV treatment.

Serum Lipids Profiling Perturbances in Patients with Ischemic Heart Disease and Ischemic Cardiomyopathy

2020 ◽  
Author(s):  
Lin Yang ◽  
Liang Wang ◽  
Yangyang Deng ◽  
Lizhe Sun ◽  
Bowen Low ◽  
...  
Keyword(s):  
Heart Disease ◽  
Ischemic Heart Disease ◽  
Ischemic Cardiomyopathy ◽  
Serum Lipids ◽  
Ischemic Heart ◽  
Negative Ion ◽  
Serum Samples ◽  
Cross Sectional ◽  
Negative Ion Mode ◽  
Positive Ion

Abstract Background: Ischemic heart disease (IHD) is a common cardiovascular disorder associated with inadequate blood supply to the myocardium. Chronic coronary ischemia leads to ischemic cardiomyopathy (ICM). Despite their rising prevalence and morbidity, few studies have discussed the lipids alterations in these patients. Methods: In this cross-sectional study, we analyzed serum lipids profile in IHD and ICM patients using a lipidomics approach. Consecutive consenting patients admitted to the hospital for IHD and ICM were enrolled. Serum samples were obtained after overnight fasting. Non-targeted metabolomics was applied to demonstrate lipids metabolic profile in control, IHD and ICM patients. Results: A total of 63 and 62 lipids were detected in negative and positive ion mode respectively. Among them, 16:0 Lyso PI, 18:1 Lyso PI in negative ion mode, and 19:0 Lyso PC, 12:0 SM d18:1/12:0, 15:0 Lyso PC, 17:0 PC, 18:1-18:0 PC in positive ion mode were significantly altered both in IHD and ICM as compared to control. 13:0 Lyso PI, 18:0 Lyso PI, 16:0 PE, 14:0 PC DMPC, 16:0 ceramide, 18:0 ceramide in negative ion mode, and 17:0 PE, 19:0 PC, 14:0 Lyso PC, 20:0 Lyso PC, 18:0 PC DSPC, 18:0-22:6 PC in positive ion mode were significantly altered only in ICM as compared to IHD and control. Conclusion: Using non-targeted lipidomics profiling, we have successfully identified a group of circulating lipids that were significantly altered in IHD and ICM. The lipids metabolic signatures shed light on potential new biomarkers and therapeutics for preventing and treating ICM.

scholarly journals HPTLC-DESI-HRMS-Based Profiling of Anthraquinones in Complex Mixtures—A Proof-of-Concept Study Using Crude Extracts of Chilean Mushrooms

Foods ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 156 ◽  
Author(s):  
Annegret Laub ◽  
Ann-Katrin Sendatzki ◽  
Götz Palfner ◽  
Ludger A. Wessjohann ◽  
Jürgen Schmidt ◽  
...  
Keyword(s):  
High Performance ◽  
High Resolution Mass Spectrometry ◽  
Desorption Electrospray Ionization ◽  
Negative Ion ◽  
Fragmentation Pattern ◽  
Proof Of Concept ◽  
Concept Study ◽  
Crude Extracts ◽  
Marker Compounds ◽  
Characteristic Fragmentation

High-performance thin-layer chromatography (HPTLC) coupled with negative ion desorption electrospray ionization high-resolution mass spectrometry (DESI-HRMS) was used for the analysis of anthraquinones in complex crude extracts of Chilean dermocyboid Cortinarii. For this proof-of-concept study, the known anthraquinones emodin, physcion, endocrocin, dermolutein, hypericin, and skyrin were identified by their elemental composition. HRMS also allowed the differentiation of the investigated anthraquinones from accompanying compounds with the same nominal mass in the crude extracts. An investigation of the characteristic fragmentation pattern of skyrin in comparison with a reference compound showed, exemplarily, the feasibility of the method for the determination of these coloring, bioactive and chemotaxonomically important marker compounds. Accordingly, we demonstrate that the coupling of HPTLC with DESI-HRMS represents an advanced and efficient technique for the detection of anthraquinones in complex matrices. This analytical approach may be applied in the field of anthraquinone-containing food and plants such as Rheum spp. (rhubarb), Aloe spp., Morinda spp., Cassia spp. and others. Furthermore, the described method can be suitable for the analysis of anthraquinone-based colorants and dyes, which are used in the food, cosmetic, and pharmaceutical industry.

Structural investigation of perfluorocarboxylic acid derivatives formed in the reaction with N,N-dimethylformamide dialkylacetals

2019 ◽  
Vol 26 (2) ◽  
pp. 131-143 ◽  
Author(s):  
Monika Stróżyńska ◽  
Jürgen H. Gross ◽  
Katrin Schuhen
Keyword(s):  
Structural Investigation ◽  
Direct Analysis ◽  
Dimethyl Acetal ◽  
Negative Ion ◽  
Cluster Ions ◽  
Perfluorocarboxylic Acid ◽  
Reaction Products ◽  
Salt Structure ◽  
Negative Ion Mode ◽  
Positive Ion

A structural investigation of perfluorocarboxylic acid derivatives formed in the reaction with N,N-dimethylformamide dialkylacetals employing several techniques of mass spectrometry (MS) is described. Two derivatizing reagents, dimethylformamide dimethyl acetal (DMF-DMA) and dimethylformamide diethylacetal (DMF-DEA) were used. In contrast to carboxylic acids, perfluorocarboxylic acids are not able to form alkyl esters as the main product in this reaction. We found that perfluorooctanoic acid (PFOA) forms a salt with N,N-dimethylformamide dialkylacetals. This salt undergoes a further reaction inside the injection block of a gas chromatograph (GC) by loss of CO2 and then forms 1,1-perfluorooctane-(N,N,N,N-tetramethyl)-diamine. The GC-MS experiments using both electron ionization (EI) and positive-ion chemical ionization (PCI) revealed that the same reaction products are formed with either derivatizing reagent. Subjecting the perfluorocarboxylic acid derivative to electrospray ionization (ESI) and direct analysis in real time (DART), both positive- and negative-ion modes indicated that cluster ions are formed. In the positive-ion mode, this cluster ion consists of two iminium cations and one PFOA anion, while in the negative-ion mode, it comprises two PFOA anions and one cation. The salt structure was further confirmed by liquid injection field desorption/ionization (LIFDI) as well as infrared (IR) spectroscopy. We propose a simple mechanism of N,N,N′,N′-tetramethylformamidinium cation formation. The structure elucidation is supported by specific fragment ions as obtained by GC-EI-MS and GC-PCI-MS analyses.

Identification of Aflatoxins by Quadrupole Mass Spectrometry/Mass Spectrometry

1984 ◽  
Vol 67 (4) ◽  
pp. 734-738 ◽  
Author(s):  
Ronald D Plattner ◽  
Glenn A Bennett ◽  
Robert D Stubblefield
Keyword(s):  
Mass Spectrometry ◽  
Aflatoxin B1 ◽  
Impact Ionization ◽  
Negative Ion ◽  
Quadrupole Mass ◽  
Molecular Anion ◽  
Negative Ion Mode ◽  
Protonated Molecules ◽  
Positive Ion ◽  
Mass Spectrometry Mass Spectrometry

Abstract MS/MS daughter experiments were recorded for aflatoxins B1, B2, G1, G2, M1, M2, and aflatoxicol, using 3 ionization modes. Daughters were recorded from the molecular ion (M+) using electron impact ionization (EI). Daughters from the protonated molecules (MH+) were recorded in the positive ion mode and the daughters from the molecular anion (M+) were recorded in the negative ion mode using chemical ionization (CI). These daughter spectra are all relatively simple. The EI daughters are quite similar to conventional EI spectra. The yield of (M-) is about 100 times greater than the yield of M+ in EI or MH+ in isobutane CI spectrum. Negative ion daughter spectra were used to demonstrate the feasibility of determining the presence of aflatoxin B1 in crude extracts of contaminated corn. Aflatoxin B1 could be detected at 10 ppb.

scholarly journals Comparative transcriptome and metabolome analysis of Ostrinia furnacalis female adults under UV-A exposure

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Li Su ◽  
Changli Yang ◽  
Jianyu Meng ◽  
Lv Zhou ◽  
Changyu Zhang
Keyword(s):  
Immune Defense ◽  
Negative Ion ◽  
Comparative Transcriptome ◽  
Ostrinia Furnacalis ◽  
Metabolome Analysis ◽  
Dna Libraries ◽  
Important Pest ◽  
Negative Ion Mode ◽  
And Control ◽  
Positive Ion

AbstractUltraviolet A (UV-A) radiation is a significant environmental factor that causes photoreceptor damage, apoptosis, and oxidative stress in insects. Ostrinia furnacalis is an important pest of corn. To understand the adaptation mechanisms of insect response to UV-A exposure, this study revealed differentially expressed genes (DEGs) and differently expressed metabolites (DEMs) in O. furnacalis under UV-A exposure. Three complementary DNA libraries were constructed from O. furnacalis adult females (CK, UV1h, and UV2h), and 50,106 expressed genes were obtained through Illumina sequencing. Of these, 157 and 637 DEGs were detected in UV1h and UV2h after UV-A exposure for 1 and 2 h, respectively, compared to CK, with 103 and 444 upregulated and 54 and 193 downregulated genes, respectively. Forty four DEGs were detected in UV2h compared to UV1h. Comparative transcriptome analysis between UV-treated and control groups revealed signal transduction, detoxification and stress response, immune defense, and antioxidative system involvement. Metabolomics analysis showed that 181 (UV1h vs. CK), 111 (UV2h vs. CK), and 34 (UV2h vs. UV1h) DEMs were obtained in positive ion mode, while 135 (UV1h vs. CK), 93 (UV2h vs. CK), and 36 (UV2h vs. UV1h) DEMs were obtained in negative ion mode. Moreover, UV-A exposure disturbed amino acid, sugar, and lipid metabolism. These findings provide insight for further studies on how insects protect themselves under UV-A stress.

scholarly journals Evaluation of 6 MALDI-Matrices for 10 μm lipid imaging and on-tissue MSn with AP-MALDI-Orbitrap

2021 ◽  
Author(s):  
Tina B. Angerer ◽  
Jerome Bour ◽  
Jean-Luc Biagi ◽  
Eugene Moskovets ◽  
Gilles Frache
Keyword(s):  
Spatial Resolution ◽  
Tissue Sample ◽  
Additional Treatment ◽  
Negative Ion ◽  
High Mass ◽  
Lipid Species ◽  
High Mass Resolution ◽  
Matrix Additives ◽  
Negative Ion Mode ◽  
Positive Ion

Mass spectrometry imaging (MSI) is a technique uniquely suited to localize and identify lipids in a tissue sample. Using an AP-MALDI UHR source coupled to an Orbitrap Elite, numerous lipid locations and structures can be determined in high mass resolution spectra and at cellular spatial resolution, but careful sample preparation is necessary. We tested 11 protocols on serial brain sections for the commonly used MALDI matrices, CHCA, Norharmane, DHB, DHAP, THAP, and DAN, in combination with tissue washing and matrix additives, to determine the lipid coverage, signal intensity, and spatial resolution achievable with AP-MALDI. In positive ion mode, the most lipids could be detected with CHCA and THAP, while THAP and DAN without additional treatment offered the best signal intensities. In negative ion mode, DAN showed the best lipid coverage and DHAP performed superior for Gangliosides. DHB produced intense cholesterol signals in the white matter. 155 lipids were assigned in positive (THAP), 137 in negative ion mode (DAN) and 76 lipids were identified using on tissue tandem-MS. The spatial resolution achievable with DAN was 10 μm, confirmed with on tissue line-scans. This enabled the association of lipid species to single neurons in AP-MALDI images. The results show that the performance of AP-MALDI is comparable to vacuum MALDI techniques for lipid imaging.

scholarly journals Microevolution in the pansecondary metabolome of Aspergillus flavus and its potential macroevolutionary implications for filamentous fungi

2021 ◽  
Vol 118 (21) ◽  
pp. e2021683118
Author(s):  
Milton T. Drott ◽  
Tomás A. Rush ◽  
Tatum R. Satterlee ◽  
Richard J. Giannone ◽  
Paul E. Abraham ◽  
...  
Keyword(s):  
Secondary Metabolism ◽  
Aspergillus Flavus ◽  
High Performance ◽  
High Resolution Mass Spectrometry ◽  
Population Level ◽  
Gene Clusters ◽  
The United States ◽  
Ecological Interactions ◽  
Different Populations ◽  
Fungal Secondary Metabolism

Fungi produce a wealth of pharmacologically bioactive secondary metabolites (SMs) from biosynthetic gene clusters (BGCs). It is common practice for drug discovery efforts to treat species’ secondary metabolomes as being well represented by a single or a small number of representative genomes. However, this approach misses the possibility that intraspecific population dynamics, such as adaptation to environmental conditions or local microbiomes, may harbor novel BGCs that contribute to the overall niche breadth of species. Using 94 isolates of Aspergillus flavus, a cosmopolitan model fungus, sampled from seven states in the United States, we dereplicate 7,821 BGCs into 92 unique BGCs. We find that more than 25% of pangenomic BGCs show population-specific patterns of presence/absence or protein divergence. Population-specific BGCs make up most of the accessory-genome BGCs, suggesting that different ecological forces that maintain accessory genomes may be partially mediated by population-specific differences in secondary metabolism. We use ultra-high-performance high-resolution mass spectrometry to confirm that these genetic differences in BGCs also result in chemotypic differences in SM production in different populations, which could mediate ecological interactions and be acted on by selection. Thus, our results suggest a paradigm shift that previously unrealized population-level reservoirs of SM diversity may be of significant evolutionary, ecological, and pharmacological importance. Last, we find that several population-specific BGCs from A. flavus are present in Aspergillus parasiticus and Aspergillus minisclerotigenes and discuss how the microevolutionary patterns we uncover inform macroevolutionary inferences and help to align fungal secondary metabolism with existing evolutionary theory.

229 RAPID, UNTARGETED LIPID DETERMINATION IN INDIVIDUAL BOVINE OOCYTES AND PRE-IMPLANTATION EMBRYOS BY HIGH-RESOLUTION DESORPTION ELECTROSPRAY IONIZATION MASS SPECTROMETRY

2013 ◽  
Vol 25 (1) ◽  
pp. 262 ◽  
Author(s):  
A. F. González-Serrano ◽  
C. R. Ferreira ◽  
V. Pirro ◽  
L. S. Eberlin ◽  
J. Heinzmann ◽  
...  
Keyword(s):  
High Resolution ◽  
Desorption Electrospray Ionization ◽  
Fatty Acyl ◽  
Glass Slide ◽  
Negative Ion ◽  
Ionization Mass ◽  
Desi Ms ◽  
Negative Ion Mode ◽  
Bovine Oocytes ◽  
Positive Ion

Lipid structural analysis in individual pre-implantation mammalian embryos is hampered by the small amount of biological material, such that most studies use staining methods or gas chromatography analysis generate information only on the fatty acyl residues. Recent developments in high-resolution desorption electrospray ionization mass spectrometry (DESI-MS) allow the analysis of free fatty acids (FA) and glycerophospholipids (PL) in individual bovine embryos. Here, we report on the use of DESI-MS for the sensitive analysis of triacylglycerol (TAG) species, profiles of FA and PL in individual bovine oocytes and embryos. Bovine oocytes (n = 40) and blastocysts (n = 42) were frozen in a minimal volume of PBS (2 to 5 µL). Samples were directly deposited on glass slides after thawing. After drying, a volume of 500 µL of methanol : water (1 : 1, vol/vol) was carefully deposited on the surface of the glass slide and removed by orienting the glass slide vertically to eliminate PBS salts. An Orbitrap mass spectrometer was used for the experiments. Parameters for the positive ion mode were as follows: acetonitrile (ACN) supplemented with 3 µL mL–1 of AgNO3 at a 5 µL min–1 flow rate, injection time of 1000 ms, and a mass-to-charge range of m/z 400 to 1500. For the negative ion mode, the solvent combination used was acetonitrile + dimethylformamide (1 : 1, vol/vol) at a 1.0 µL min–1 flow rate, a maximum injection time of 1000 ms, and a mass-to-charge range of m/z 150 to 1000. Positive ion mode data for the detection of TAG species were acquired first, followed by acquisition of FA and PL in the negative ion mode. Detection of TAG by DESI, which is extremely useful for bovine embryo cryopreservation and metabolism research, has been performed by adding AgNO3 in the DESI spray to obtain silver adducts, which are easily recognised by the characteristic 1 : 1 abundance ratio of the 107 : 109 Ag isotopes. The most abundant fatty acyl residues present in TAG species were palmitic (P), linoleic (L), oleic (O), and stearic (S) acids, such as TAG of m/z 937, PPL (50 : 2); m/z 965, POO (52 : 1); m/z 967, POS (52 : 2); m/z 989, OOL/LLS (54 : 4); and m/z 991, OOO, SOL (54 : 3). Free FA and PL profiles collected from the same samples in the negative ion mode were similar to those in our recent report (2012 J. Mass Spectrom. 47, 29–33). Lipid attribution has been performed based on high-resolution mass analysis. Multivariate statistics from this data set will allow visualisation of differences observed in the lipid profiles among samples. In conclusion, we report the use of DESI-MS for the sensitive analysis of TAG in individual bovine oocytes and embryos and the creation of profiles of FA, PL, and TAG species in the same sample by DESI-MS.

1 An HPLC-MS/MS Method Using a Multitoxin Clean-up Column for Analysis of Seven Mycotoxins in Aquafeeds

2021 ◽  
Author(s):  
Siyuan Bi ◽  
Jingbing Xu ◽  
Xiaoshan Yang ◽  
Peng Zhang ◽  
Kaoqi Lian ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Performance ◽  
Correlation Coefficients ◽  
Negative Ion ◽  
Aflatoxin M1 ◽  
Contamination Level ◽  
Ionization Mass ◽  
External Standard Method ◽  
The Matrix ◽  
Negative Ion Mode

Abstract Background In Guangdong Province of China, the climate here is very wet, so there are many different fungus living in the aquatic feeds, which produce mycotoxins. These compounds contaminate agriculture products world-wide and represent a great threat to human health. It is necessary to determine their contamination level in aquatic feeds. Objective A high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method was developed for the quantitative analysis of aflatoxin B1, aflatoxin M1, T-2 toxin, HT-2 toxin, deoxynivalenol, ochratoxin, and zearalenone in the fish and shrimp feed. Methods Samples were extracted with acetonitrile-water (V: V = 3:1), and degreased with acetonitrile-saturated hexane. Such obtained extract was cleaned up with a multitoxin column. The target compounds were separated on a C18 chromatographic column and analyzed simultaneously by electrospray ionization mass spectrometry in both positive or negative ion mode. Detected compounds were quantified by using the matrix-matched external standard method. Results Under the optimized conditions, good linearities for the analytes in corresponding concentration range were obtained with correlation coefficients (r2) higher than 0.9948. LOD ranged from 1.83 to 12.63 μg/kg, and LOQ ranged from 5.49 to 37.89 μg/kg. Average recoveries for the target mycotoxins at three spiked levels ranged from 80.5% to 116.5% with RSD ranging from 2.4% to 10.4%. 23 real aquafeed samples were determined by this method, and 7 kinds of toxins were all detected. Conclusions Obtained results showed that developed method could be successfully applied for the simultaneous determination of mycotoxins in aquatic feeds.

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