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Establishment of a targeted analysis method for gangliosides in mouse tissues by HILIC-ESI–MS/MS

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Abstract

Gangliosides play an imperative role in cell signaling, neuronal recovery, apoptosis, and other physiological processes. For example, GM3 can regulate hypothalamic leptin resistance and control energy homeostasis, GD3 can mediate cell proliferation and differentiation and induce apoptosis, and GQ1b can stimulate neurogenesis. Therefore, the present study sought to establish and optimize the targeted analysis method for ganglioside subclasses and their molecular species using hydrophilic interaction liquid chromatography–triple quadrupole–MS/MS (HILIC-QQQ-MS/MS). Additionally, the fragmentation pattern of different ganglioside subclasses and their retention time patterns were analyzed, providing more accurate qualitative results. The limit of quantitation (LOQ) was as low as 10−4 ng. Moreover, the molecular species of gangliosides in the liver, cortex, and hypothalamus of C57BL/6 mice were analyzed using the established method. A total of 23 ganglioside subclasses with 164 molecular species, including 40 O-acetylated ganglioside molecular species and 28 NeuGc ganglioside molecular species, were identified using the semi-quantitative analysis method of an external standard curve corrected by an internal standard. In addition to NeuGc gangliosides, the contents of ganglioside subclasses were more abundant in the mouse brain than those in the mouse liver; especially, the contents of unsaturated gangliosides in the hypothalamus were much higher than those in the liver. Among them, O-acetylated gangliosides were detected only in the cortex and hypothalamus at a concentration of up to 100 μg/mg protein (40 molecular species). Overall, the proposed method expanded the detectable number of ganglioside subclasses and molecular species in biological samples and provided more opportunities for further study of the biological functions of gangliosides.

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Abbreviations

Cer:

Ceramide

Glc:

D-glucose

Gal:

D-galactose

GalNAc:

N-acetylgalactosamine

Fuc:

Fucose

NeuAc:

N-acetylneuraminic acid

NeuGc:

N-glycolylneuraminic acid

O-Ac:

O-Acetylated

GC-MS:

Gas chromatography–mass spectrometry

AP-MALDI-MS:

Atmospheric pressure matrix-assisted laser desorption ionization mass spectrometry

HILIC-ESI–MS/MS:

Hydrophilic interaction liquid chromatography–electrospray ionization coupled with mass spectrometry

MRM:

Multi-stage reaction monitoring

QQQ:

Triple quadrupole

QC:

Quality control

LOD:

Limit of detection

LOQ:

Limit of quantification

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Funding

This work was supported by the Fundamental Research Funds for the Central Universities of China (No. 202261047 and No. 202012018).

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Authors and Affiliations

  1. State Key Laboratory of Marine Food Processing & Safety Control, College of Food Science and Engineering, Ocean University of China, No. 1299, Sansha Road, Qingdao, 266404, Shandong, China

    Shuo Yang, Yingxu Ma, Yu Song, Xiaoxu Wang, Peixu Cong, Nan Meng, Jie Xu & Changhu Xue

  2. Qingdao Marine Science and Technology Center, Qingdao, 266235, China

    Changhu Xue

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  1. Shuo Yang
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Correspondence to Peixu Cong or Jie Xu.

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The animal experiment involved in our investigation was approved and conducted following the Animal Ethics Committee of the Ocean University of China (Approval No. SPXY2022062801).

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Published in the topical collection New Trends in Lipidomics with guest editor Michal Holčapek.

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Yang, S., Ma, Y., Song, Y. et al. Establishment of a targeted analysis method for gangliosides in mouse tissues by HILIC-ESI–MS/MS. Anal Bioanal Chem (2024). https://doi.org/10.1007/s00216-024-05169-0

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