TitleUntargeted Identification of Alkyne-Containing Natural Products Using Ruthenium-Catalyzed Azide Alkyne Cycloaddition Reactions Coupled to LC-MS/MS.
Publication TypeJournal Article
Year of Publication2022
AuthorsBack D, Shaffer BT, Loper JE, Philmus B
JournalJ Nat Prod
Volume85
Issue1
Pagination105-114
Date Published2022 01 28
ISSN1520-6025
KeywordsAlkynes, Biological Products, Catalysis, Chromatography, Liquid, Cycloaddition Reaction, Ruthenium, Tandem Mass Spectrometry
Abstract

Alkyne-containing natural products have been identified from plants, insects, algae, fungi, and bacteria. This class of natural products has been characterized as having a variety of biological activities. Polyynes are a subclass of acetylenic natural products that contain conjugated alkynes and are underrepresented in natural product databases due to the fact that they decompose during purification. Here we report a workflow that utilizes alkyne azide cycloaddition (AAC) reactions followed by LC-MS/MS analysis to identify acetylenic natural products. In this report, we demonstrate that alkyne azide cycloaddition reactions with -bromobenzyl azide result in -bromobenzyl-substituted triazole products that fragment to a common brominated tropylium ion. We were able to identify a synthetic alkyne spiked into the extract of sp. PCC 7120 at a concentration of 10 μg/mL after optimization of MS/MS conditions. We then successfully identified the known natural product fischerellin A in the extract of PCC 9339. Lastly, we identified the recently identified natural products protegenins A and C from Pf-5 through a combination of genome mining and RuAAC reactions. This is the first report of RuAAC reactions to detect acetylenic natural products. We also compare CuAAC and RuAAC reactions and find that CuAAC reactions produce fewer byproducts compared to RuAAC but is limited to terminal-alkyne-containing compounds. In contrast, RuAAC is capable of identification of both terminal and internal acetylenic natural products, but byproducts need to be eliminated from analysis by creation of an exclusion list. We believe that both CuAAC and RuAAC reactions coupled to LC-MS/MS represent a method for the untargeted identification of acetylenic natural products, but each method has strengths and weaknesses.

DOI10.1021/acs.jnatprod.1c00798
Alternate JournalJ Nat Prod
PubMed ID35044192
PubMed Central IDPMC8853637
Grant ListR15 GM117541 / GM / NIGMS NIH HHS / United States
S10 OD018518 / OD / NIH HHS / United States
T32 AT010131 / AT / NCCIH NIH HHS / United States