Quantitative Analysis of Lipids: A Higher-Throughput LC–MS/MS-Based Method and its Comparison to ELISA

Abstract

Aim: Lipids such as prostaglandins, leukotrienes and thromboxanes are released as a result of an inflammatory episode in pain (central and peripheral). Methodology & results: To measure these lipids as potential mechanistic biomarkers in neuropathic pain models, we developed a higher-throughput LC–MS/MS-based method with simultaneous detection of PGE2, PGD2, PGF2α, LTB4, TXB2 and 2-arachidonoyl glycerol in brain and spinal cord tissues. We also demonstrate that the LC–MS/MS method was more sensitive and specific in differentiating PGE2 levels in CNS tissues compared with ELISA. Conclusion: The ability to modify the LC–MS/MS method to accommodate numerous other lipids in one analysis, demonstrates that the presented method offers a cost–effective and more sensitive alternative to ELISA method useful in drug discovery settings.

Lay abstract In humans, lipids carry out various functions such as energy production and storage, insulation, digestion and absorption and hormone production. Out of the several lipids, prostaglandins, thromboxanes and leukotrienes play a critical role in cardiovascular diseases, allergic reactions and inflammation. Thus, it is important to monitor their levels as potential mechanistic biomarkers to effectively diagnose and treat the underlying diseases. We have successfully used a highly specific and higher-throughput mass spectrometric method to quantify these lipids in brain cells as well as in brain and spinal cord tissues from rats (pain model) and compared the data obtained in the traditional ELISA.

Keywords::

• ELISA
• higher-throughput
• LC–MS/MS
• lipids
• neuropathic pain

Author contributions

AS Gandhi, R Staal and G Chandrasena participated in research design. AS Gandhi and T Khayrullina conducted experiments. R Staal and G Chandrasena contributed new reagents or analytic tools. AS Gandhi, D Budac, T Khayrullina, R Staal and G Chandrasena performed data analysis. AS Gandhi, D Budac, T Khayrullina, R Staal and G Chandrasena wrote or contributed to the writing of the manuscript.

Acknowledgements

The authors thank the research management team at Lundbeck Research USA for their valuable inputs in reviewing the manuscript. The authors would also like to thank Manuel Cajina for his technical assistance.

Financial & competing interests disclosure

This work was supported by internal budget of Lundbeck Research Inc. USA as part of the CNS drug discovery program. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

This work was supported by internal budget of Lundbeck Research Inc. USA as part of the CNS drug discovery program. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.