Journal
JOURNAL OF NATURAL PRODUCTS
Volume 78, Issue 8, Pages 1926-1936Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jnatprod.5b00268
Keywords
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Funding
- National Cancer Institute/National Institutes of Health, Bethesda, MD, USA [P01 CA125066]
- North Carolina Biotechnology Center [2011-BRG-1206]
- Cooperative Research and Development Agreement (CRADA) [NFE-10-02966]
- AB Sciex
- U.S. Department of Energy [DE-AC05-00OR22725]
- NATIONAL CANCER INSTITUTE [P01CA125066] Funding Source: NIH RePORTER
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Ambient ionization mass spectrometry techniques have recently become prevalent in natural product research due to their ability to examine secondary metabolites in situ. These techniques retain invaluable spatial and temporal details that are lost through traditional extraction processes. However, most ambient ionization techniques do not collect mutually supportive data, such as chromatographic retention times and/or UV/vis spectra, and this can limit the ability to identify certain metabolites, such as differentiating isomers. To overcome this, the droplet liquid microjunction-surface sampling probe (droplet-LMJ-SSP) was coupled with UPLC-PDA-HRMS-MS/MS, thus providing separation, retention times, MS data, and UV/vis data used in traditional dereplication protocols. By capturing these mutually supportive data, the identity of secondary metabolites can be confidently and rapidly assigned in situ. Using the droplet-LMJ-SSP, a protocol was constructed to analyze the secondary metabolite profile of fungal cultures without any sample preparation. The results demonstrate that fungal cultures can be dereplicated from the Petri dish, thus identifying secondary metabolites, including isomers, and confirming them against reference standards. Furthermore, heat maps, similar to mass spectrometry imaging, can be used to ascertain the location and relative concentration of secondary metabolites directly on the surface and/or surroundings of a fungal culture.
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