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Quantification of sialic acids in red meat by UPLC-FLD using indoxylsialosides as internal standards

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Abstract

Herein we describe a UPLC-FLD-based method for the quantification of the sialic acid content of red meat, using a synthetic neuraminic acid derivative as an internal standard. X-Gal-α-2,6-N-propionylneuraminic acid was synthesized via a chemoenzymatic pathway and its hydrolytic stability was characterized. Known quantities of this compound were incubated with samples of red meat under sialic acid-releasing conditions. The released sialic acids were derivatized, analyzed by UPLC-FLD, and the Neu5Ac/Neu5Gc content of the meat sample was determined by comparison with the internal standard. A number of red meats were analyzed by this method with the following results (Neu5Ac μg/g tissue, Neu5Gc μg/g tissue ± s.d.): pork (68 ± 3, 15.2 ± 0.7), beef (69 ± 8, 36 ± 5), lamb (46 ± 2, 33 ± 1), rabbit (59 ± 2, 0.4 ± 0.4), and hare (50 ± 4, 1 ± 1). We envisage that this methodology will find application in investigating the health effects of dietary Neu5Gc.

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Abbreviations

CMP:

cytidyl 5′-monophosphate

DCC:

dicyclohexylcarbodimide

EcNeuS:

Escherichia coli neuraminic acid synthase

ESI:

electrospray ionization

FLD:

fluorescence detector

Kdn:

α-2,6-keto-3-deoxy-d-glycero-d-galactononic acid

MALDI:

matrix-assisted laser desorption ionization

MS:

mass spectrometry

Neu5Ac:

N-acetylneuraminic acid

Neu5Gc:

N-glycolylneuraminic acid

Neu5Pr:

N-propylneuraminic acid

NHS:

N-hydroxysuccinimide

NmCSS:

Neisseria meningitidis CMP-sialic acid synthase

PdST6:

Photobacterium damselae α-2,6-sialyltransferase

OPD:

o-phenylenediamine

UPLC:

ultra high performance liquid chromatography

X-Gal:

5-bromo-4-chloro-indolyl-β-d-galactopyranoside

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Acknowledgments

The authors would like to thank Prof. Akemi Suzuki (Hiratsuka, Japan) for helpful discussions on sialic acids.

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Authors and Affiliations

  1. Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, China

    Hong L. Yao, Louis P. Conway, Mao M. Wang, Kun Huang, Li Liu & Josef Voglmeir

  2. Food Science Department, Jingling Institute of Technology, Nanjing, China

    Hong L. Yao

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  1. Hong L. Yao
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  2. Louis P. Conway
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  3. Mao M. Wang
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  4. Kun Huang
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  5. Li Liu
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  6. Josef Voglmeir
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Correspondence to Li Liu or Josef Voglmeir.

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Source of funding

This work was supported in parts by the Natural Science Foundation of China (grant number 31,471,703 to L.L. and J.V., A0201300537 to J.V. and L.L.), Natural Science Foundation of the Jiangsu Province Higher Education Institutions (grant number 13KJD230002 to L.H.Y.), and the 100 Foreign Talents Plan (grant number JSB2014012 to J.V.).

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Yao, H.L., Conway, L.P., Wang, M.M. et al. Quantification of sialic acids in red meat by UPLC-FLD using indoxylsialosides as internal standards. Glycoconj J 33, 219–226 (2016). https://doi.org/10.1007/s10719-016-9659-1

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