UPLC-Q-TOF-MS/MS-Based Targeted Discovery of Chetomin Analogues from Chaetomium cochliodes.

Chetocochliodin M (5) containing a rare cage-ring and chetocochliodin N (6) featuring an unusual piperazine-2,3-dione ring system together with known analogues chetomin (1), chetoseminudin C (2), chetocochliodin I (3), and oidioperazine E (4) were targeted for purification from the fungus Chaetomium cochliodes using a UPLC-Q-TOF-MS/MS approach. The structures of the new compounds were elucidated using HR-ESI-MS, NMR, and ECD spectra. Compounds 1, 3, and 6 exhibited strong cytotoxic activities against A549 and HeLa cancer cell lines.

Bioactive secondary metabolites from endophytic strains of Neocamarosporium betae collected from desert plants.

Endophytic fungi from desert plants belong to a unique microbial community that has been scarcely investigated chemically and could be a new resource for bioactive natural products. In this study, 13 secondary metabolites (1-13) with diverse carbon skeletons, including a novel polyketide (1) with a unique 5,6-dihydro-4H,7H-2,6-methanopyrano[4,3-d][1,3]dioxocin-7-one ring system and three undescribed polyketides (2, 7, and 11), were obtained from the endophytic fungus Neocamarosporium betae isolated from two desert plant species. Different approaches, including HR-ESI-MS, UV spectroscopy, IR spectroscopy, NMR, and CD, were used to determine the planar and absolute configurations of the compounds. The possible biosynthetic pathways were proposed based on the structural characteristics of compounds 1-13. Compounds 1, 3, 4, and 9 exhibited strong cytotoxicity toward HepG2 cells compared with the positive control. Several metabolites (2, 4-5, 7-9, and 11-13) were phytotoxic to foxtail leaves. The results support the hypothesis that endophytic fungi from special environments, such as desert areas, produce novel bioactive secondary metabolites.

Undescribed diphenyl ethers betaethrins A-I from a desert plant endophytic strain of the fungus Phoma betae A.B. Frank (Didymellaceae).

Ten diphenyl ethers (DPEs), including nine undescribed analogs named betaethrins A-I, were isolated from the desert plant endophytic fungus Phoma betae A.B. Frank (Didymellaceae). Their structures were determined mainly by NMR, HR-ESI-MS spectral and X-ray diffraction experiments. Betaethrins D-I possessed different fatty acid chains connected with the B-ring, which was the first report in all DPEs. The shielding effect of the B-ring on H-6 (A-ring) in methyl barceloneate, betaethrin A and betaethrins D-F (asterric acid analogs) was first observed and analyzed, which could differentiate the 1H-NMR chemical shift values of H-4/H-6 without the assistance of 3-OH. An empirical rule was then suggested: the steric hindrance between the A- and B-rings in asterric acid analogs might prevent these two aromatic rings from rotating freely, which led to the 1H-NMR chemical shift value of H-6 being in the high field zone due to the shielding effect of the B-ring on H-6. Based on the empirical rule, the chemical shift values of the A-ring in methyl barceloneate were revised. The possible biosynthesis of these isolates was postulated. Betaethrin H showed moderate cytotoxicity against MCF-7 and HepG2 cancer cell lines. Betaethrins A-F, H and I displayed strong antioxidant activities. These results further implied that endophytic fungi from unique environments, such as desert plants, with few chemical studies are an important resource of undescribed and bioactive metabolites.

UPLC-Q-TOF-MS/MS Analysis of Seco-Sativene Sesquiterpenoids to Detect New and Bioactive Analogues From Plant Pathogen Bipolaris sorokiniana.

Seco-sativene sesquiterpenoids are an important member of phytotoxins and plant growth regulators isolated from a narrow spectrum of fungi. In this report, eight seco-sativene sesquiterpenoids (1-8) were first analyzed using the UPLC-Q-TOF-MS/MS technique in positive mode, from which their mass fragmentation pathways were suggested. McLafferty rearrangement, 1,3-rearrangement, and neutral losses were considered to be the main fragmentation patterns for the [M+1]+ ions of 1-8. According to the structural features (of different substitutes at C-1, C-2, and C-13) in compounds 1-8, five subtypes (A-E) of seco-sativene were suggested, from which subtypes A, B/D, and E possessed the diagnostic daughter ions at m/z 175, 189, and 203, respectively, whereas subtype C had the characteristic daughter ion at m/z 187 in the UPLC-Q-TOF-MS/MS profiles. Based on the fragmentation patterns of 1-8, several known compounds (1-8) and two new analogues (9 and 10) were detected in the extract of plant pathogen fungus Bipolaris sorokiniana based on UPLC-Q-TOF-MS/MS analysis, of which 1, 2, 9, and 10 were then isolated and elucidated by NMR spectra. The UPLC-Q-TOF-MS/MS spectra of these two new compounds (9 and 10) were consistent with the fragmentation mechanisms of 1-8. Compound 1 displayed moderate antioxidant activities with IC50 of 0.90 and 1.97 mM for DPPH and ABTS+ scavenging capacity, respectively. The results demonstrated that seco-sativene sesquiterpenoids with the same subtypes possessed the same diagnostic daughter ions in the UPLC-Q-TOF-MS/MS profiles, which could contribute to structural characterization of seco-sativene sesquiterpenoids. Our results also further supported that UPLC-Q-TOF-MS/MS is a powerful and sensitive tool for dereplication and detection of new analogues from crude extracts of different biological origins.

UPLC-Q-TOF-MS/MS analysis of the guaiane sesquiterpenoids oxytropiols A-J and detection of undescribed analogues from the locoweed endophytic fungus Alternaria oxytropis (Pleosporaceae).

INTRODUCTION: In addition to the mycotoxin swainsonine, the locoweed endophytic fungus Alternaria oxytropis (Pleosporaceae) also produces a series of rarely reported, highly oxygenated bicyclic guaiane sesquiterpenoids. Few investigations on the electrospray tandem mass fragmentation pattern of this sesquiterpenoid have been reported. OBJECTIVES: We aimed to analyze and detect new guaiane sesquiterpenoid analogues from crude extracts of the locoweed endophytic fungus A. oxytropis by UPLC-Q-TOF-MS/MS experiments. MATERIALS AND METHODS: Oxytropiols A-J (1-10) and the extract of the locoweed endophytic fungus A. oxytropis were analyzed by UPLC-Q-TOF-MS/MS in positive mode. RESULTS: Typical neutral losses, McLafferty rearrangement, 1,2-rearrangement, and 1,3-rearrangement were considered to be the main fragmentation patterns for the [M + H]+ /[M + Na]+ ions of 1-10 by UPLC-Q-TOF-MS/MS experiments, and possible fragmentation pathways of 1-10 were suggested. A unique and undescribed analogue named oxytropiol K (11) was found in the extract based on UPLC-Q-TOF-MS/MS analysis. Compound 11 was isolated and elucidated by NMR spectrometry, and its UPLC-Q-TOF-MS/MS analysis was consistent with the fragmentation pathways of 1-10. CONCLUSION: The results further support that UPLC-Q-TOF-MS/MS is a powerful and sensitive tool for the characterization of known compounds (dereplication) and the detection of new analogues from crude extracts and imply that the locoweed endophytic fungus A. oxytropis, with few chemical investigations, is an important resource for undescribed metabolites.

Accurate Identification of Degraded Products of Aflatoxin B1 Under UV Irradiation Based on UPLC-Q-TOF-MS/MS and NMR Analysis.

Analysis, purification, and characterization of AFB1 degraded products are vital steps for elucidation of the photocatalytic mechanism. In this report, the UPLC-Q-TOF-MS/MS technique was first coupled with purification and NMR spectral approaches to analyze and characterize degraded products of AFB1 photocatalyzed under UV irradiation. A total of seventeen degraded products were characterized based on the UPLC-Q-TOF-MS/MS analysis, in which seven ones (1-7) including four (stereo) isomers (1,2, 5, and 6) were purified and elucidated by NMR experiments. According to the structural features of AFB1 and degraded products (1-7), the possible photocatalytic mechanisms were suggested. Furthermore, AFB1 and degraded products (1-7) were evaluated against different cell lines. The results indicated that the UPLC-Q-TOF-MS/MS technique combined with purification, NMR spectral experiments, and biological tests was an applicable integrated approach for analysis, characterization, and toxic evaluation of degraded products of AFB1, which could be used to evaluate other mycotoxin degradation processes.

Bioactive seco-Sativene Sesquiterpenoids from an Artemisia desertorum Endophytic Fungus, Cochliobolus sativus.

A series of seco-sativene sesquiterpenoids (1-11) including two new natural products (2 and 3), four new analogues (4-7), and six known analogues, helminthosporic acid (1), drechslerine A (8), drechslerine B (9), helminthosporol (10), helminthosporal acid (11), and isosativenediol (12), were purified from the endophytic fungus Cochliobolus sativus isolated from a desert plant, Artemisia desertorum. The stereochemistry of helminthosporic acid (1) was established for the first time by X-ray diffraction, and the structures including relative and absolute configurations of these new compounds were determined by NMR and CD spectra together with biosynthetic considerations. Compounds 5-7 are the first seco-sativene sesquiterpenoids possessing a glucose group on C-15, C-15, and C-14, respectively. Compounds 1, 7, 9, and 11 displayed strong phytotoxic effects on corn leaves by producing visible lesions, and helminthosporic acid (1) was shown to promote division of leaves and roots of Arabidopsis thaliana with a dose-dependent relationship.

Revision of the structure of isochaetoglobosin Db based on NMR analysis and biosynthetic consideration.

Isochaetoglobosin Db is a new chaetoglobosin possessing a unique 3,4-substituted pyrrole ring isolated and named by Qiu et al., and it is different from any one of the 14 sub-types in the macrocyclic ring of chaetoglobosins classified in our previous work. Its chemical shift values, coupling constants and biosynthetic consideration implied that the proposed structure of isochaetoglobosin Db was incorrect. In this report, based on detailed NMR data analysis together with biosynthetic consideration, the structure of isochaetoglobosin Db is suggested to be revised to that of penochalasin C. The NMR spectra of penochalasin C measured in the same solvent (DMSO-d 6) as that of isochaetoglobosin Db supported the above conclusion. The results imply that reasonable biosynthetic consideration could complement spectroscopic structural determination, and also support that the 1H-NMR rule of chaetoglobosin summarized in our previous work can provide help for dereplication and rectification.