Advertisement

Analytical and Bioanalytical Chemistry

, Volume 411, Issue 6, pp 1239–1251 | Cite as

Reversed phase UHPLC/ESI-MS determination of oxylipins in human plasma: a case study of female breast cancer

  • Michaela Chocholoušková
  • Robert Jirásko
  • David Vrána
  • Jiří Gatěk
  • Bohuslav Melichar
  • Michal HolčapekEmail author
Research Paper

Abstract

The ultrahigh-performance liquid chromatography-mass spectrometry (UHPLC/MS) method was optimized and validated for the determination of oxylipins in human plasma using the targeted approach with selected reaction monitoring (SRM) in the negative-ion electrospray ionization (ESI) mode. Reversed phase UHPLC separation on an octadecylsilica column enabled the analysis of 63 oxylipins including numerous isomeric species within 12-min run time. The method was validated (calibration curve, linearity, limit of detection, limit of quantification, carry-over, precision, accuracy, recovery rate, and matrix effect) and applied to 40 human female plasma samples from breast cancer patients and age-matched healthy volunteers (control). Thirty-six oxylipins were detected in human plasma with concentrations above the limit of detection, and 21 of them were quantified with concentrations above the limit of quantitation. The concentrations determined in healthy controls are in a good agreement with previously reported data on human plasma. Quantitative data were statistically evaluated by multivariate data analysis (MDA) methods including principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA). S-plot and box plots showed that 13-HODE, 9-HODE, 13-HOTrE, 9-HOTrE, and 12-HHTrE were the most upregulated oxylipin species in plasma of breast cancer patients.

Keywords

Oxylipins Eicosanoids UHPLC/MS Breast cancer Statistical analysis Human plasma 

Notes

Acknowledgments

We would like to acknowledge the help of Tereza Hrnčiarová with the statistical analysis.

Funding information

This work was supported by ERC CZ project No. LL1302 sponsored by the Ministry of Education, Youth and Sports of the Czech Republic.

Compliance with ethical standards

The study was approved by the institutional ethical committee. All patients and healthy volunteers signed informed consent.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1556_MOESM1_ESM.pdf (9.4 mb)
ESM 1 (PDF 9.44 MB)
216_2018_1556_MOESM2_ESM.xlsx (23 kb)
ESM 2 (XLSX 22 kb)

References

  1. 1.
    Pelengaris S, Khan M. The molecular biology of cancer: a bridge from bench to bedside. 2nd ed. Oxford: Wiley-Blackwell; 2013.Google Scholar
  2. 2.
    Weiss MC. Breast cancer information and support [Internet]. Breast cancer.org. 2018. Available from: https://www.breastcancer.org/. Accessed 26 July 2018.
  3. 3.
    Melichar B. Laboratory medicine and medical oncology: the tale of two Cinderellas. Clin Chem Lab Med. 2013;51:99–112.Google Scholar
  4. 4.
    Melichar B, Hornychova H, Kalabova H, Basova H, Mergancova J, Urminska H, et al. Increased efficacy of a dose-dense regimen of neoadjuvant chemotherapy in breast carcinoma: a retrospective analysis. Med Oncol. 2012;29:2577–85.Google Scholar
  5. 5.
    Cifkova E, Lisa M, Hrstka R, Vrana D, Gatek J, Melichar B, et al. Correlation of lipidomic composition of cell lines and tissues of breast cancer patients using hydrophilic interaction liquid chromatography/electrospray ionization mass spectrometry and multivariate data analysis. Rapid Commun Mass Spectrom. 2017;21:253–63.Google Scholar
  6. 6.
    Cifkova E, Holcapek M, Lisa M, Vrana D, Gatek J, Melichar B. Determination of lipidomic differences between human breast cancer and surrounding normal tissues using HILIC-HPLC/ESI-MS and multivariate data analysis. Anal Bioanal Chem. 2015;407:991–1002.Google Scholar
  7. 7.
    Jirasko R, Holcapek M, Khalikova M, Vrana D, Student V, Prouzova Z, et al. MALDI orbitrap mass spectrometry profiling of dysregulated sulfoglycosphingolipids in renal cell carcinoma tissues. J Am Soc Mass Spectrom. 2017;28:1562–74.Google Scholar
  8. 8.
    Hajek R, Lisa M, Khalikova M, Jirasko R, Cifkova E, Student V, et al. HILIC/ESI-MS determination of gangliosides and other polar lipid classes in renal cell carcinoma and surrounding normal tissues. Anal Bioanal Chem. 2018;410:6585–94.Google Scholar
  9. 9.
    Melichar B, Konopleva M, Hu W, Melicharova K, Andreeff M, Freedman RS. Growth-inhibitory effect of a novel synthetic triterpenoid, 2-cyano-3,12-dioxoolean-1,9-dien-oic acid, on ovarian carcinoma cell lines not dependent on peroxisome proliferator-activated receptor-g expression. Gynecol Oncol. 2004;93:149–54.Google Scholar
  10. 10.
    Krop IE, Mayer IA, Ganju V, Dickler M, Johnston S, Morales S, et al. Pictilisib for estrogen receptor-positive, aromatase inhibitor-resistant, advanced or metastatic breast cancer (FERGI): a randomised, double-blind, placebo-controlled trial. Lancet Oncol. 2016;17:811–21.Google Scholar
  11. 11.
    Berkecz R, Lisa M, Holcapek M. Analysis of oxylipins in human plasma: comparison of utrahigh-performance liquid chromatography and ultrahigh-performance supercritical fluid chromatography coupled to mass spectrometry. J Chromatogr A. 2017;1511:107–21.Google Scholar
  12. 12.
    Strassburg K, Huijbrechts AML, Kortekaas KA, Lindeman JH, Pedersen TL, Dane A, et al. Quantitative profiling of oxylipins through comprehensive LC-MS/MS analysis: application in cardiac surgery. Anal Bioanal Chem. 2012;404:1413–26.Google Scholar
  13. 13.
    Wang Y, Armando AM, Quehenberger O, Yan C, Dennis EA. Comprehensive ultra-performance liquid chromatographic separation and mass spectrometric analysis of eicosanoids metabolites in human samples. J Chromatogr A. 2017;1359:60–9.Google Scholar
  14. 14.
    Kortz L, Dorow J, Becker S, Thiery J, Ceglarek U. Fast liquid chromatography-quadrupole linear ion trap-mass spectrometry analysis of polyunsaturated fatty acids and eicosanoids in human plasma. J Chromatogr B. 2013;927:209–13.Google Scholar
  15. 15.
    Gabbs M, Leng S, Devassy JG, Monirujjaman M, Aukema HM. Advances in our understanding of oxylipins derived from dietary PUFAs. Adv Nutr. 2015;6:513–40.Google Scholar
  16. 16.
    Schebb NH, Ostermann AI, Yang J, Hammock BD, Hahn A, Schuchardt JP. Comparison of the effects of long-chain omega-3 fatty acid supplementation on plasma levels of free and esterified oxylipins. Prostaglandins Other Lipid Mediat. 2014;113-115:21–9.Google Scholar
  17. 17.
    Yamada H, Oshiro E, Kikuchi S, Hakozaki M, Takahashi H, Kinura K. Hydroxyeicosapentaenoic acids from the Pacific krill show high ligand activities for PPARs. J Lipid Res. 2014;55:895–904.Google Scholar
  18. 18.
    Massey KA, Nicolaou A. Lipidomics of oxidized polyunsaturated fatty acids. Free Radic Biol Med. 2013;59:45–55.Google Scholar
  19. 19.
    Shinde DD, Kim KB, Oh KS, Abdalla N, Liu KH, Bae SK, et al. LC-MS/MS for the simultaneous analysis of arachidonic acid and 32 related metabolites in human plasma: basal plasma concentrations and aspirin-induced changes of eicosanoids. J Chromatogr B. 2012;911:113–21.Google Scholar
  20. 20.
    Song J, Liu X, Wu J, Meehan MJ, Blevitt JM, Dorrestein PC, et al. A highly efficient, high-throughput lipidomics platform for the quantitative detection of eicosanoids in human whole blood. Anal Biochem. 2013;433:181–8.Google Scholar
  21. 21.
    Buczynski MW, Dumlao DS, Dennis EA. Thematic review series: proteomics. An integrated omics analysis of eicosanoids biology. J Lipid Res. 2009;50:1015–38.Google Scholar
  22. 22.
    Burke JE, Dennis EA. Phospholipase A2 structure/function, mechanism, and signaling. J Lipid Res. 2009;50(Suppl):S237–42.Google Scholar
  23. 23.
    Komaba JJ, Matsuda D, Shibakawa K, Nakade S, Hashimoto Y, Miyata Y, et al. Development and validation of an online-two-dimensional reversed-phase liquid chromatography-tandem mass spectrometry method for the simultaneous determination of prostaglandin E2, F2α, and 13,14-dihydro-15-keto prostaglandin F2α levels in human plasma. Biomed Chromatogr. 2009;23:315–23.Google Scholar
  24. 24.
    Shono F, Yokota K, Horie K, Yamamoto S, Yamashita K, Watanabe K, et al. A heterologous enzyme immunoassay of prostaglandin E2 using a stable enzyme-labeled hapten mimic. Anal Biochem. 1988;168:284–91.Google Scholar
  25. 25.
    Reinke M. Monitoring thromboxane in body fluids: a specific ELISA for 11-dihydrothromboxane B2 using a monoclonal antibody. Am J Physiol Endocrinol Metab. 1992;262:E658–62.Google Scholar
  26. 26.
    Tsikas D. Application of gas chromatography-mass spectrometry and gas chromatography-tandem mass spectrometry to assess in vivo synthesis of prostaglandins, thromboxane, leukotrienes, isoprostanes and related compounds in humans. J Chromatogr B. 1998;717:201–45.Google Scholar
  27. 27.
    Agency EMA. Committee for Medicinal Products for Human Use (CHMP) [Internet]. European medicines agency. 2011. Available from: https://www.ema.europa.eu/en/committees/committee-medicinal-products-human-use-chm. Accessed 10 July 2018.
  28. 28.
    Food U, Administration D. US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research: Rockville. 2001.Google Scholar
  29. 29.
    The R foundation. The R project for statistical computing [Internet]. R-project.org. 2018. Available from: https://www.r-project.org/. Accessed 26 July 2018.
  30. 30.
    Blanchard J. Evaluation of the relative efficacy of various techniques for deproteinizing plasma samples prior to high-performance liquid chromatographic analysis. J Chromatogr B. 1981;226:455–60.Google Scholar
  31. 31.
    Cifkova E, Hajek R, Lisa M, Holcapek M. Hydrophilic interaction liquid chromatography-mass spectrometry of (lyso)phosphatidics, (lyso)phosphatidylserines and other lipid classes. J Chromatogr A. 2016;1439:65–73.Google Scholar
  32. 32.
    Loomba R, Quehenberger O, Armando A, Dennis EA. Polyunsaturated fatty acid metabolites as novel lipidomic biomarkers for noninvasive diagnosis of nonalcoholic steatohepatitis. J Lipid Res. 2015;56:185–92.Google Scholar
  33. 33.
    Quehenberger O, Armando AM, Brown AH, Milne SB, Myers DS, Merrill AH, et al. Lipidomics reveals a remarkable diversity of lipids in human plasma. J Lipid Res. 2010;51:3299–305.Google Scholar
  34. 34.
    Ferreiro-Vera C, Mata-Granados JM, Priego-Capote F, Quesada-Gómez JM, Luque de Castro MD. Automated targeting analysis of eicosanoids inflammation biomarkers in human serum and in the exometabolome of stem cells by SPE-LC-MS/MS. Anal Bioanal Chem. 2011;399:1093–103.Google Scholar
  35. 35.
    Schuchardt JP, Schmidt S, Kressel G, Dong H, Willenberg I, Hammock BD, et al. Comparison of free serum oxylipin concentrations in hyper- vs. normolipidemic men. Prostaglandins Leukot Essent Fatty Acids. 2013;89:19–29.Google Scholar
  36. 36.
    Schuchardt JP, Schmidt S, Kressel G, Dong H, Willenberg I, Hammock BD, et al. Modulation of blood oxylipin levels by long-chain omega-3 fatty acid supplementation in hyper- and normolipidemic men. Prostaglandins Leukot Essent Fatty Acids. 2014;90:27–37.Google Scholar
  37. 37.
    Psychogios N, Hau DD, Peng J, Guo AC, Mandal R, Bouatra S, et al. The human serum metabolome. PLoS One. 2011;6:e16957.Google Scholar
  38. 38.
    Caligiuri SPB, Aukema HM, Ravandi A, Pierce GN. Elevated levels of pro-inflammatory oxylipins in older subjects are normalized by flaxseed consumption. Exp Gerontol. 2014;59:51–7.Google Scholar
  39. 39.
    Bowden JA, et al. Harmonizing lipidomics: NIST interlaboratory comparison exercise for lipidomics using standard reference material 1950 – metabolites in frozen human plasma. J Lipid Res. 2017;58:2275–88.Google Scholar
  40. 40.
    Yuan ZX, Majchrzak-Hong S, Keyes GS, Iadarola MJ, Mannes AJ, Ramsden CE. Lipidomic profiling of targeted oxylipins with ultra-performance liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem. 2018;140:6009–29.Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Michaela Chocholoušková
    • 1
  • Robert Jirásko
    • 1
  • David Vrána
    • 2
  • Jiří Gatěk
    • 3
  • Bohuslav Melichar
    • 2
  • Michal Holčapek
    • 1
    Email author
  1. 1.Faculty of Chemical Technology, Department of Analytical ChemistryUniversity of PardubicePardubiceCzech Republic
  2. 2.Medical School and Teaching Hospital, Department of OncologyPalacký UniversityOlomoucCzech Republic
  3. 3.Atlas Hospital, Department of SurgeryTomáš Baťa University in ZlínZlínCzech Republic

Personalised recommendations