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Comprehensive Analysis of Different Subtypes of Oxylipins to Determine a LC-MS/MS Approach in Clinical Research
Yurou Zhao1, Zhengyu Fang1, Zeyu Li1
1Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Background/Objectives:
Different oxylipin subtypes have unique biological properties, requiring effective analytical protocols. However, establishing a complete pathway detection protocol for comprehensive oxylipin analysis is challenging. This study aimed to evaluate the adaptability and specificity of oxylipin subtypes under different extraction schemes and to develop a robust analytical platform for clinical biomarker investigation.
Methods:
We revealed the adaptability and specificity of oxylipin subtypes based on different single-step extraction schemes. A high-throughput quantitative automated solid-phase extraction coupled with a liquid chromatography-tandem mass spectrometry (aSPE-LC-MS/MS) analytical platform was established for a broad panel of complex oxylipins. The method was applied to serum samples of patients with coronary heart disease (CHD).
Results:
Our results verified that oxo-oxylipins, resolvin, and eicosanoids showed the best extraction efficiency under SPE protocol. Most hydroxy-oxylipins, dihydroxy-oxylipins, and HOTrEs are suitable for methanol protocol, HDHA for acetonitrile protocol, and epoxy-oxylipins for the methyl tert-butyl ether protocol, while medium-chain HETE is suitable for ethyl acetate protocol. Importantly, a novel sensitive fast method with wide coverage by the aSPE-LC-MS/MS analytical platform with satisfying sensitivity, accuracy and precision, extraction efficiency, low matrix effect, and linear calibration curves was obtained. Furthermore, we have successfully applied this method and found that 5-HETE, 11-HETE, and 15-HETE can serve as integrated biomarkers for patients with CHD, with high diagnostic performance.
Conclusions:
The study provides the best protocol for the clinically targeted detection of oxylipins and provides an important means for studying biomarkers of diseases.
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