A liquid chromatography tandem mass spectrometry-based method for the quantitation of 39 lipid mediators in four sample types and in two mouse strains is described. The method builds upon existing methodologies for analysis of lipid mediators by A) utilizing a bead homogenization step for tissue samples; this eliminates the need for homogenization glassware and improves homogenization consistency, B) optimizing the isolation and purification of lipid mediators with polymeric reverse phase SPE columns with lower sorbent masses; this results in lower solvent elution volumes without loss of recovery and C) utilizing an on-column enrichment method to improve analyte focusing before chromatographic separation. The method is linear from 0.25-250 pg on column for low level lipid mediators and from 5-5000 pg on column for high level lipid mediators. The addition of a methyl formate elution step to a previously published method dramatically improved precision and recovery for the cysteinyl leukotrienes. Accuracy and precision for 4 different sample types including human plasma, mouse lung, mouse spleen and mouse liver is demonstrated. Liver samples had extremely high levels of a tentatively identified bile acid which interfered with quantitation of resolvin E1, 11B-prostaglandin F2a and thromboxane A2. Results from 2 different tissue sources from untreated mice (C57BL/6 versus BALB/c) showed dramatically different concentrations of lipid mediators.

描述了一种基于液相色谱串联质谱的方法，用于定量四种样品类型和两种小鼠品系中的 39 种脂质介质。该方法建立在现有的脂质介质分析方法的基础上，A) 利用组织样品的珠子均质化步骤；这消除了对均质玻璃器皿的需求并提高了均质一致性，B) 使用具有较低吸附剂质量的聚合物反相 SPE 柱优化脂质介质的分离和纯化；这导致较低的溶剂洗脱体积而不损失回收率和 C) 在色谱分离之前利用柱上富集方法改善分析物聚焦。该方法线性范围为 0.25-250 pg 的低浓度脂质介体和 5-5000 pg 的高浓度脂质介体。在先前发表的方法中增加甲酸甲酯洗脱步骤，显着提高了半胱氨酰白三烯的精确度和回收率。证明了 4 种不同样品类型的准确度和精密度，包括人血浆、小鼠肺、小鼠脾和小鼠肝脏。肝脏样本中一种初步鉴定的胆汁酸含量极高，这会干扰溶解素 E1、11B-前列腺素 F2a 和血栓素 A2 的定量。来自未治疗小鼠的 2 种不同组织来源（C57BL/6 与 BALB/c）的结果显示脂质介质的浓度显着不同。演示了小鼠脾脏和小鼠肝脏。肝脏样本中一种初步鉴定的胆汁酸含量极高，这会干扰溶解素 E1、11B-前列腺素 F2a 和血栓素 A2 的定量。来自未治疗小鼠的 2 种不同组织来源（C57BL/6 与 BALB/c）的结果显示脂质介质的浓度显着不同。演示了小鼠脾脏和小鼠肝脏。肝脏样本中一种初步鉴定的胆汁酸含量极高，这会干扰溶解素 E1、11B-前列腺素 F2a 和血栓素 A2 的定量。来自未治疗小鼠的 2 种不同组织来源（C57BL/6 与 BALB/c）的结果显示脂质介质的浓度显着不同。