Rapid Screening of Ellagitannins in Natural Sources via Targeted Reporter Ion Triggered Tandem Mass Spectrometry

Complex biomolecules present in their natural sources have been difficult to analyze using traditional analytical approaches. Ultrahigh-performance liquid chromatography (UHPLC-MS/MS) methods have the potential to enhance the discovery of a less well characterized and challenging class of biomolecules in plants, the ellagitannins. We present an approach that allows for the screening of ellagitannins by employing higher energy collision dissociation (HCD) to generate reporter ions for classification and collision-induced dissociation (CID) to generate unique fragmentation spectra for isomeric variants of previously unreported species. Ellagitannin anions efficiently form three characteristic reporter ions after HCD fragmentation that allows for the classification of unknown precursors that we call targeted reporter ion triggering (TRT). We demonstrate how a tandem HCD-CID experiment might be used to screen natural sources using UHPLC-MS/MS by application of 22 method conditions from which an optimized data-dependent acquisition (DDA) emerged. The method was verified not to yield false-positive results in complex plant matrices. We were able to identify 154 non-isomeric ellagitannins from strawberry leaves, which is 17 times higher than previously reported in the same matrix. The systematic inclusion of CID spectra for isomers of each species classified as an ellagitannin has never been possible before the development of this approach

Bispericyclic Diels-Alder Dimerization of ortho-Quinols in Natural Product (Bio)Synthesis -Bioinspired Chemical 6-Step Synthesis of (+)-Maytenone

International audienc

Infused Grandinin MS-MS

The MS/MS files for the grandinin ellagitannin standard

Custom Perl Script

<div>The custom perl script used to extract precursors subjected to CID or HCD from mgf files at the end of the proteome discoverer workflow.</div><div><br></div

Infused Roburin D MS-MS

The MS/MS files for the roburin D ellagitannin standard

Infused Castalagin MS-MS

The MS/MS files for the castalagin ellagitannin standard

UHPLC HCD-CID Instrument Iterations, CID

<p>Systematic variation of the AGC target and CID resolution to determine the effect on total HCD-CID screening output with other instrumental parameters fixed.</p

Infused Roburin BC MS-MS

The MS/MS files for the roburin BC ellagitannin mixed standard

Infused Epiacutissimin A MS-MS

The MS/MS files for the epiacutissimin A ellagitannin standard

UHPLC False Positive Controls, Amaranth

Application of different charge state and reporter ion triggering conditions on amaranth control samples, ellagitannin standards, and a mixture of the ellagitannin standards and the amaranth control