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Rapid, sensitive, and high-throughput quantification of broad serological ceramides by using isotope dilution liquid chromatography-negative ion electrospray tandem mass spectrometry

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Abstract

Ceramides are important intermediates in the metabolism of sphingolipids. High-throughput liquid chromatography-mass spectrometry has been used extensively for monitoring the levels of serological ceramides, but is still limited by inadequate coverage or lack of sensitivity. Herein, a rapid, sensitive, and high-throughput isotope dilution liquid chromatography-negative ion electrospray tandem mass spectrometry (IDLC-nESI-MS/MS) method was developed and verified for accurate quantification of 41 ceramides, involving ceramides with C16-20 sphingosine, dihydro-ceramide, and dehydro-ceramide. This method was validated with excellent linearity (R2 > 0.99) and good recovery in the range of 90–110%. Intra- and inter-day imprecision were below 5.57% and 7.83% respectively. The improved high-throughput quantitative method developed in this study may aid in the accurate characterization of ceramides for understanding ceramide biology and application in disease diagnosis.

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Acknowledgements

The authors would like to acknowledge the financial support from the National Natural Science Foundation of China (grant no. 21904058).

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Authors and Affiliations

  1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, 110016, Liaoning, China

    Shuailong Chen, Longshan Zhao & Zhili Xiong

  2. School of Medicine, Academy for Advanced Interdisciplinary Studies, Southern University of Science and Technology, 1088 Xueyuan Rd., Shenzhen, China

    Shuailong Chen, Xuan Li, Wenyong Zhang & Hemi Luan

  3. Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China

    Shijia Liu

  4. State Key Laboratory of Chemical Oncogenomics, Tsinghua Shenzhen International Graduate School, Shenzhen, PR China

    Hemi Luan

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  1. Shuailong Chen
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  2. Xuan Li
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  4. Longshan Zhao
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Correspondence to Longshan Zhao, Wenyong Zhang, Zhili Xiong or Hemi Luan.

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This study was approved by the medical ethics committee of the Affiliated Hospital of Nanjing University of Chinese Medicine and followed the declaration of Helsinki (2018NL-106-02).

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Chen, S., Li, X., Liu, S. et al. Rapid, sensitive, and high-throughput quantification of broad serological ceramides by using isotope dilution liquid chromatography-negative ion electrospray tandem mass spectrometry. Anal Bioanal Chem 415, 801–808 (2023). https://doi.org/10.1007/s00216-022-04473-x

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