Determination of endogenous inflammation-related lipid mediators in ischemic stroke rats using background subtracting calibration curves by liquid chromatography-tandem mass spectrometry.

Yang, Yang; Zhong, Qisheng; Mo, Canlong; Zhang, Hao; Zhou, Ting; Tan, Wen

Accurate and reliable quantification of endogenous lipid mediators in complex biological samples is a daunting challenge. In this study, a robust and direct endogenous quantitative method using background subtracting calibration curves by liquid chromatography-tandem mass spectrometry was first developed for the determination of endogenous lipid mediators in ischemic stroke rats. Absolute quantification without surrogate matrix could be achieved by using background subtracting calibration curves, which were corrected and verified from standard curves constructed on original matrix. The recoveries of this method were in the range of 50.3-98.3%, the precision with the relative standard deviation was less than 13.8%, and the accuracy with the relative error was within ± 15.0%. In addition, background subtracting calibration curves were further verified by validation factors ranging from 90.3 to 110.9%. This validated method has been successfully applied to the analysis of seven endogenous inflammation-related lipid mediators in the brain tissues of ischemic stroke rats. The results indicated that prostaglandins as inflammatory factors and some lipid mediators with neuroprotective effects increased apparently ( p < 0.05) in the stroke groups compared with the normal rats. Besides, the two drugs (isosteviol sodium and edaravone) could significantly reduce ( p < 0.05) the levels of prostaglandin E and prostaglandin F of stroke rats to inhibit inflammation. Based on the results, it is strongly believed that this approach can be readily generalized as a new reference for the quantification of endogenous compounds in the complex biological samples.

STROKE treatment; NEUROPROTECTIVE agents; LIQUID chromatography; TANDEM mass spectrometry; PROSTAGLANDINS; CEREBROVASCULAR disease; MULTI-infarct dementia

Analytical & Bioanalytical Chemistry, 2017, Vol 409, Issue 28, p6537

1618-2642

Academic Journal

10.1007/s00216-017-0600-7