Yang Yang, Zhong Qisheng, Mo Canlong, Zhang Hao, Zhou Ting, Tan Wen
School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong, 510006, China.
Shimadzu (China) Corporation, Guangzhou, Guangdong, 510010, China.
Anal Bioanal Chem. 2017 Nov;409(28):6537-6547. doi: 10.1007/s00216-017-0600-7. Epub 2017 Sep 4.
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. Graphical abstract The analysis procedure of determining endogenous inflammation-related lipid mediators using BSCC by LC-MS/MS.
对复杂生物样品中的内源性脂质介质进行准确可靠的定量分析是一项艰巨的挑战。在本研究中，首次开发了一种稳健且直接的内源性定量方法，即利用液相色谱 - 串联质谱法通过背景扣除校准曲线来测定缺血性中风大鼠体内的内源性脂质介质。通过使用背景扣除校准曲线可以实现无需替代基质的绝对定量，该曲线由在原始基质上构建的标准曲线进行校正和验证。该方法的回收率在50.3 - 98.3%范围内，相对标准偏差表示的精密度小于13.8%，相对误差表示的准确度在±15.0%以内。此外，通过90.3%至110.9%的验证因子进一步验证了背景扣除校准曲线。这种经过验证的方法已成功应用于缺血性中风大鼠脑组织中七种内源性炎症相关脂质介质的分析。结果表明，与正常大鼠相比，作为炎症因子的前列腺素和一些具有神经保护作用的脂质介质在中风组中明显增加（p < 0.05）。此外，两种药物（异甜菊醇钠和依达拉奉）可显著降低（p < 0.05）中风大鼠的前列腺素E和前列腺素F水平以抑制炎症。基于这些结果，坚信这种方法可以很容易地推广，作为复杂生物样品中内源性化合物定量分析的新参考。图形摘要 使用液相色谱 - 串联质谱法通过背景扣除校准曲线测定内源性炎症相关脂质介质的分析程序。
