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Improved specificity of serum phosphatidylcholine detection based on side-chain losses during negative electrospray ionization tandem mass spectrometry

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Abstract

Many current tandem mass spectrometry (MS) methods for measuring phosphatidylcholines (PtdChos) rely only on precursor ion scanning of the common 184 m/z phosphocholine fragment with positive electrospray ionization (+ESI), and thus measure pools of PtdChos rather than specific isoforms. In this paper, we developed and compared an isotope dilution, tandem MS method capable of quantifying PtdChos based on specific fatty acid side-chains to the traditional 184 m/z method. The method is based on the detection of PtdCho ammonium formate (AmF) adduct as parent ions and fatty acid fragment daughter ions under negative electrospray ionization (−ESI). Accuracy, imprecision, and recovery were below 15 %, with acceptable linearity (R 2 > 0.99) up to 5 μg/mL. We used the method to analyze the distributions of PtdChos with common side-chain combinations among 60 subjects and showed that it was possible for two individuals to have the same PtdCho pool concentration based on detection of the 184 m/z fragment, but up to a fourfold difference in the levels of specific isoforms comprising the pool based on our method. We then compared the results of both methods across 572 patients with mild cognitive impairment (MCI), Alzheimer’s disease (AD), or no impairment (NI), which showed that statistically significant associations between specific PtdCho isoforms and AD were masked with the 184 m/z method. Our findings demonstrate the importance of isoform specificity for quantifying PtdChos, and suggest caution when interpreting analytical data based on pools of biomarkers.

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Acknowledgments

We acknowledge Precision Medicine for providing the Alzheimer’s disease samples used in this study, and Dr. Vijitha Senanayake for his careful review of the manuscript.

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Authors and Affiliations

  1. Phenomenome Discoveries Inc., 204-407 Downey Road, Saskatoon, Saskatchewan, S7N 4L8, Canada

    Shawn A Ritchie, Dushmanthi Jayasinge, Li Wang & Dayan B Goodenowe

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  1. Shawn A Ritchie
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  3. Li Wang
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Correspondence to Shawn A Ritchie.

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Conflict of interest

All authors were employees of, and received salaries from, Phenomenome Discoveries, Inc.

Samples from patients with Alzheimer’s disease (AD, n = 204) and mild cognitive impairment (MCI, n = 210) and no cognitive impairment (NC, n = 158) were provided by Precision Medicine Inc (USA) according to the criteria from the National Institute of Neurological and Communicative Diseases and Stroke/Alzheimer’s Disease and Related Disorders Association (NINCDS-ADRDA). All samples were collected under informed consent.

All studies described in this manuscript were approved by the appropriate ethics committee and were performed in accordance with ethical standards.

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Ritchie, S.A., Jayasinge, D., Wang, L. et al. Improved specificity of serum phosphatidylcholine detection based on side-chain losses during negative electrospray ionization tandem mass spectrometry. Anal Bioanal Chem 408, 7811–7823 (2016). https://doi.org/10.1007/s00216-016-9884-2

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  • DOI: https://doi.org/10.1007/s00216-016-9884-2

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