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Development, validation and comparison of three LC-MS/MS methods for determination of endogenous striatal oleoyl ethanolamine in mice.
Journal of Chromatography B ( IF 2.8 ) Pub Date : 2020-02-24 , DOI: 10.1016/j.jchromb.2020.122041 Yeli Guan 1 , Qiao Deng 2 , Gao Li 1 , Luqin Si 1 , Lihong Long 2 , Mohammadali Soleimani Damaneh 1 , Jiangeng Huang 1
Journal of Chromatography B ( IF 2.8 ) Pub Date : 2020-02-24 , DOI: 10.1016/j.jchromb.2020.122041 Yeli Guan 1 , Qiao Deng 2 , Gao Li 1 , Luqin Si 1 , Lihong Long 2 , Mohammadali Soleimani Damaneh 1 , Jiangeng Huang 1
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Obesity has become a severe public health problem worldwide. An endogenous fatty acid ethanolamine oleoyl ethanolamine (OEA) is reported to be capable of reducing body weight and food intake by increasing striatal extracellular dopamine concentration. However, association between obesity and striatal OEA level remains unknown. As such, it is necessary to develop a sensitive and reliable method to quantitate OEA concentration in striatum. Because true endogenous analytes free blank matrix is not available, surrogate analyte, surrogate matrix and background subtraction methods are often employed for the analysis of endogenous compounds. In this study, three liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were developed and validated for the determination of OEA concentration in mouse brain homogenate. Interestingly, stability results found that OEA-d4 degraded in brain homogenate under room temperature, while OEA level remarkably increased with time. Since lowering temperature could observably decelerate the endogenous transformation of OEA, sample collection and preparation were carried out under ice-bath condition. Hexane: isopropanol (9:1, v/v) was employed as an extractant for liquid-liquid extraction. After method validation, three methods were applied to quantify OEA in striatum homogenate from C57B6/L mice following normal and high fat diet feeding for 4 months. Results from three methods all showed the striatal OEA level in obesity group was significantly higher than control group and obesity-resist group, which indicated that obesity might be associated with elevated striatal OEA level.
中文翻译：
三种LC-MS / MS测定，测定小鼠内源性纹状体油酰乙醇胺的方法的开发，验证和比较。
肥胖已成为世界范围内严重的公共卫生问题。据报道，内源性脂肪酸乙醇胺油酰乙醇胺（OEA）能够通过增加纹状体细胞外多巴胺浓度来减轻体重和食物摄入。但是，肥胖与纹状体OEA水平之间的关联仍然未知。因此，有必要开发一种灵敏可靠的方法来定量纹状体中的OEA浓度。由于没有真正的不含内源分析物的空白基质，因此通常采用替代分析物，替代基质和背景扣除法来分析内源性化合物。在这项研究中，开发了三种液相色谱-串联质谱（LC-MS / MS）方法，并验证了该方法可用于测定小鼠脑匀浆中的OEA浓度。有趣的是 稳定性结果发现，室温下脑组织匀浆中的OEA-d4降解，而OEA水平随时间显着增加。由于降低温度可以明显地降低OEA的内源转化，因此在冰浴条件下进行样品收集和制备。己烷：异丙醇（9：1，v / v）用作液-液萃取的萃取剂。在方法验证后，采用三种方法对正常和高脂饮食喂养4个月后来自C57B6 / L小鼠的纹状体匀浆中的OEA进行定量。三种方法的结果均显示，肥胖组的纹状体OEA水平明显高于对照组和肥胖抵抗组，这表明肥胖可能与纹状体OEA水平升高有关。
更新日期：2020-02-25
中文翻译：
三种LC-MS / MS测定，测定小鼠内源性纹状体油酰乙醇胺的方法的开发，验证和比较。
肥胖已成为世界范围内严重的公共卫生问题。据报道，内源性脂肪酸乙醇胺油酰乙醇胺（OEA）能够通过增加纹状体细胞外多巴胺浓度来减轻体重和食物摄入。但是，肥胖与纹状体OEA水平之间的关联仍然未知。因此，有必要开发一种灵敏可靠的方法来定量纹状体中的OEA浓度。由于没有真正的不含内源分析物的空白基质，因此通常采用替代分析物，替代基质和背景扣除法来分析内源性化合物。在这项研究中，开发了三种液相色谱-串联质谱（LC-MS / MS）方法，并验证了该方法可用于测定小鼠脑匀浆中的OEA浓度。有趣的是 稳定性结果发现，室温下脑组织匀浆中的OEA-d4降解，而OEA水平随时间显着增加。由于降低温度可以明显地降低OEA的内源转化，因此在冰浴条件下进行样品收集和制备。己烷：异丙醇（9：1，v / v）用作液-液萃取的萃取剂。在方法验证后，采用三种方法对正常和高脂饮食喂养4个月后来自C57B6 / L小鼠的纹状体匀浆中的OEA进行定量。三种方法的结果均显示，肥胖组的纹状体OEA水平明显高于对照组和肥胖抵抗组，这表明肥胖可能与纹状体OEA水平升高有关。
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