Analytical and Bioanalytical Chemistry

, Volume 410, Issue 25, pp 6585–6594 | Cite as

HILIC/ESI-MS determination of gangliosides and other polar lipid classes in renal cell carcinoma and surrounding normal tissues

  • Roman Hájek
  • Miroslav Lísa
  • Maria Khalikova
  • Robert Jirásko
  • Eva Cífková
  • Vladimír ŠtudentJr
  • David Vrána
  • Lukáš Opálka
  • Kateřina Vávrová
  • Marcel Matzenauer
  • Bohuslav Melichar
  • Michal HolčapekEmail author
Research Paper


Negative-ion hydrophilic liquid chromatography-electrospray ionization mass spectrometry (HILIC/ESI-MS) method has been optimized for the quantitative analysis of ganglioside (GM3) and other polar lipid classes, such as sulfohexosylceramides (SulfoHexCer), sulfodihexosylceramides (SulfoHex2Cer), phosphatidylglycerols (PG), phosphatidylinositols (PI), lysophosphatidylinositols (LPI), and phosphatidylserines (PS). The method is fully validated for the quantitation of the studied lipids in kidney normal and tumor tissues of renal cell carcinoma (RCC) patients based on the lipid class separation and the coelution of lipid class internal standard with the species from the same lipid class. The raw data are semi-automatically processed using our software LipidQuant and statistically evaluated using multivariate data analysis (MDA) methods, which allows the complete differentiation of both groups with 100% specificity and sensitivity. In total, 21 GM3, 28 SulfoHexCer, 26 SulfoHex2Cer, 10 PG, 19 PI, 4 LPI, and 7 PS are determined in the aqueous phase of lipidomic extracts from kidney tumor tissue samples and surrounding normal tissue samples of 20 RCC patients. S-plots allow the identification of most upregulated (PI 40:5, PI 40:4, GM3 34:1, and GM3 42:2) and most downregulated (PI 32:0, PI 34:0, PS 36:4, and LPI 16:0) lipids, which are primarily responsible for the differentiation of tumor and normal groups. Another confirmation of most dysregulated lipids is performed by the calculation of fold changes together with T and p values to highlight their statistical significance. The comparison of HILIC/ESI-MS data and matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI) data confirms that lipid dysregulation patterns are similar for both methods.

Graphical abstract


Lipids Lipidomics Gangliosides Mass spectrometry HILIC Renal cell carcinoma Tumor tissues 



The help of Assoc. Prof. Jozef Škarda with the histological staining is gratefully acknowledged.

Funding information

The present work was supported by ERC CZ project no. LL1302 sponsored by the Ministry of Education, Youth and Sports of the Czech Republic. K.V. and L.O. thank the support of grant project no. 16-25687J sponsored by the Czech Science Foundation.

Compliance with ethical standards

The study was approved by the hospital Ethical Committee, and patients signed informed consent.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1263_MOESM1_ESM.pdf (419 kb)
ESM 1 (PDF 418 kb)
216_2018_1263_MOESM2_ESM.xlsx (83 kb)
ESM 2 (XLSX 83 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Roman Hájek
    • 1
  • Miroslav Lísa
    • 1
  • Maria Khalikova
    • 1
  • Robert Jirásko
    • 1
  • Eva Cífková
    • 1
  • Vladimír ŠtudentJr
    • 2
  • David Vrána
    • 3
  • Lukáš Opálka
    • 4
  • Kateřina Vávrová
    • 4
  • Marcel Matzenauer
    • 3
  • Bohuslav Melichar
    • 3
  • Michal Holčapek
    • 1
    Email author
  1. 1.Faculty of Chemical Technology, Department of Analytical ChemistryUniversity of PardubicePardubiceCzech Republic
  2. 2.Department of Urology, Faculty of Medicine and DentistryPalacký University and University HospitalOlomoucCzech Republic
  3. 3.Department of Oncology, Faculty of Medicine and DentistryPalacký University and University HospitalOlomoucCzech Republic
  4. 4.Faculty of Pharmacy Hradec Králové, Department of Organic and Bioorganic ChemistryCharles UniversityHradec KrálovéCzech Republic

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