Synchronized Survey Scan Approach Allows for Efficient Discrimination of Isomeric and Isobaric Compounds during LC-MS/MS Analyses

Abstract

Liquid chromatography-mass spectrometry- (LC-MS-) based multiple reaction monitoring (MRM) methods have been used to detect and quantify metabolites for years. These approaches rely on the monitoring of various fragmentation pathways of multiple precursors and the subsequent corresponding product ions. However, MRM methods are incapable of confidently discriminating between isomeric and isobaric molecules and, as such, the development of methods capable of overcoming this challenge has become imperative. Due to increasing scanning rates of recent MS instruments, it is now possible to operate MS instruments both in the static and dynamic modes. One such method is known as synchronized survey scan (SSS), which is capable of acquiring a product ion scan (PIS) during MRM analysis. The current study shows, for the first time, the use of SSS-based PIS approach as a feasible identification feature of MRM. To achieve the above, five positional isomers of dicaffeoylquinic acids (diCQAs) were studied with the aid of SSS-based PIS method. Here, the MRM transitions were automatically optimized using a 3,5-diCQA isomer by monitoring fragmentation transitions common to all five isomers. Using the mixture of these isomers, fragmentation spectra of the five isomers achieved with SSS-based PIS were used to identify each isomer based on previously published hierarchical fragmentation keys. The optimized method was also used to detect and distinguish between diCQA components found in Bidens pilosa and their isobaric counterparts found in Moringa oleifera plants. Thus, the method was shown to distinguish (by differences in fragmentation patterns) between diCQA and their isobars, caffeoylquinic acid (CQA) glycosides. In conclusion, SSS allowed the detection and discrimination of isomeric and isobaric compounds in a single chromatographic run by producing a PIS spectrum, triggered in the automatic MS/MS synchronized survey scan mode.

Detailed Phenolic Characterization of Protea Pure and Hybrid Cultivars by Liquid Chromatography–Ion Mobility–High Resolution Mass Spectrometry (LC-IM-HR-MS)

“…Advances in tandem MS technology such as the data-independent acquisition strategies allow both the precursor and product ion data to be collected in a single chromatographic run, , eliminating the requirement of multiple injections. Nevertheless, the complexity of plant samples, which often contain isomeric and/or isobaric metabolites, − poses a severe analytical challenge.…”

Active compounds from Calendula officinalis flowers act via PI3K and ERK signaling pathways to offer neuroprotective effects against Parkinson's disease

“…Chlorogenic acid (1) produced a [M − H] − ion at m/z 353.0871 and a prominent product ion at m/z 191 [quinic acid−H] − (Ye et al., 2010 ). The 3,4‐dicaffeoylquinic acid (2) produced a [M − H] − ion at m/z 515.1206 and yielded four diagnostic fragment ions at m/z 353 [caffeoylquinic acids−H − H 2 O] − , 191 [quinic acid−H] − , 179 [caffeoyl−H] − and 173 [quinic acid−H − H 2 O] − (Masike & Madala, 2018 ). Rutin (3) exhibited a [M − H] − peak at m/z 609.1461 and a major MS/MS fragment at m/z 300 (aglycone) (Jeszka‐Skowron & Zgoła‐Grześkowiak, 2014 ).…”