Profiling the Oxylipin and Endocannabinoid Metabolome by UPLC-ESI-MS/MS in Human Plasma to Monitor Postprandial Inflammation.
Profiling the Oxylipin and Endocannabinoid Metabolome by UPLC-ESI-MS/MS in Human Plasma to Monitor Postprandial Inflammation.
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在人血浆中，通过UPLC-ESI-MS/MS分析了Oxylipin和内源性大麻素的代谢组，以监测餐后炎症。
DOI：
10.1371/journal.pone.0132042
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Nording ML
中科院分区：
文献类型：
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作者：
Gouveia-Figueira S;Späth J;Zivkovic AM;Nording ML
生物活性脂质，包括氧脂素、内源性大麻素和相关化合物，可能作为炎症某些方面的特定生化标志物。然而，这些化合物的餐后反应性在很大程度上是未知的；因此，在一名自由改变日常饮食的受试者中，研究了六次应激餐后循环氧脂素和内源性大麻素代谢组的变化。饮食的改变，尤其是挑战餐本身，代表了前体脂肪酸状态的改变，预期对生物活性脂质水平有微妙的影响。为了检测最微小的变化，采用高灵敏度超高效液相色谱（UPLC）耦合电喷雾电离（ESI）串联质谱（MS/MS）方法进行生物活性脂质谱分析。之前验证UPLC-ESI-MS / MS方法分析神经代谢物,而验证UPLC-ESI-MS oxylipin / MS方法分析与可接受的结果多数参数根据美国食品和药物管理局线性指南(0.9938 < R2 < 0.9996),检测极限(0.0005 - -2.1 pg列),量化的限制(0.0005 - -4.2 pg列),国米,盘中精度(85 - 115%)和精密(< 5%),回收率40-109%，稳定性40-105%。在空腹和餐后状态（餐后0.5、1和3小时）的血浆样品中检测到52种生物活性脂质的47种。多变量分析显示，在餐后状态下，由于饮食中含有乳制品，生物活性脂质谱发生了显著变化，这与单变量分析一致，揭示了7种化合物(NAGly, 9- hode, 13-oxo- ode, 9（10)-EpOME, 12(13)-EpOME， 20-HETE和11,12- dhet）在餐后状态下的背景饮食之间存在显著差异（但在禁食时没有）。空腹时基线水平的唯一变化是TXB2。此外，还检测了7种化合物(POEA、SEA、9（10)-DiHOME、12(13)-DiHOME、13-oxo- ode、9- hode和13- hode）的餐后反应性。因此，数据证实UPLC-ESI-MS/MS方法的性能足以检测i)由饮食变化引起的餐后生物活性脂质代谢组的变化，在目前的情况下，最明显的是ii)氧化脂素和内源性大麻素代谢组亚群对挑战餐的反应性。总之，我们已经展示了UPLC-ESI-MS/MS生物活性脂质方案的概念验证，目的是监测细微的变化，从而有助于解决脂质介导的餐后炎症。
Bioactive lipids, including oxylipins, endocannabinoids, and related compounds may function as specific biochemical markers of certain aspects of inflammation. However, the postprandial responsiveness of these compounds is largely unknown; therefore, changes in the circulating oxylipin and endocannabinoid metabolome in response to a challenge meal were investigated at six occasions in a subject who freely modified her usual diet. The dietary change, and especially the challenge meal itself, represented a modification of precursor fatty acid status, with expectedly subtle effects on bioactive lipid levels. To detect even the slightest alteration, highly sensitive ultra-performance liquid chromatography (UPLC) coupled to electrospray ionization (ESI) tandem mass spectrometry (MS/MS) methods for bioactive lipid profiling was employed. A previously validated UPLC-ESI-MS/MS method for profiling the endocannabinoid metabolome was used, while validation of an UPLC-ESI-MS/MS method for oxylipin analysis was performed with acceptable outcomes for a majority of the parameters according to the US Food and Drug Administration guidelines for linearity (0.9938 < R2 < 0.9996), limit of detection (0.0005–2.1 pg on column), limit of quantification (0.0005–4.2 pg on column), inter- and intraday accuracy (85–115%) and precision (< 5%), recovery (40–109%) and stability (40–105%). Forty-seven of fifty-two bioactive lipids were detected in plasma samples at fasting and in the postprandial state (0.5, 1, and 3 hours after the meal). Multivariate analysis showed a significant shift of bioactive lipid profiles in the postprandial state due to inclusion of dairy products in the diet, which was in line with univariate analysis revealing seven compounds (NAGly, 9-HODE, 13-oxo-ODE, 9(10)-EpOME, 12(13)-EpOME, 20-HETE, and 11,12-DHET) that were significantly different between background diets in the postprandial state (but not at fasting). The only change in baseline levels at fasting was displayed by TXB2. Furthermore, postprandial responsiveness was detected for seven compounds (POEA, SEA, 9(10)-DiHOME, 12(13)-DiHOME, 13-oxo-ODE, 9-HODE, and 13-HODE). Hence, the data confirm that the UPLC-ESI-MS/MS method performance was sufficient to detect i) a shift, in the current case most notably in the postprandial bioactive lipid metabolome, caused by changes in diet and ii) responsiveness to a challenge meal for a subset of the oxylipin and endocannabinoid metabolome. To summarize, we have shown proof-of-concept of our UPLC-ESI-MS/MS bioactive lipid protocols for the purpose of monitoring subtle shifts, and thereby useful to address lipid-mediated postprandial inflammation.