Analysis of 14 endocannabinoids and endocannabinoid congeners in human plasma using column switching high-performance atmospheric pressure chemical ionization liquid chromatography-mass spectrometry.
Analysis of 14 endocannabinoids and endocannabinoid congeners in human plasma using column switching high-performance atmospheric pressure chemical ionization liquid chromatography-mass spectrometry.
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使用柱切换高性能大气压化学电离液相色谱-质谱法分析人血浆中的 14 种内源性大麻素和内源性大麻素同系物。
DOI：
10.1007/s00216-021-03280-0
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发表时间：
2021
影响因子：
4.3
通讯作者：
Klawitter,Jost
中科院分区：
文献类型：
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作者：
Sempio,Cristina;Klawitter,Jelena;Jackson,Matthew;Freni,Francesca;Shillingburg,Ryan;Hutchison,Kent;Bidwell,LCinnamon;Christians,Uwe;Klawitter,Jost
内源性大麻素系统（ECS）是一个复杂的细胞信号系统。为了满足对捕获内源性大麻素水平以研究ECS的分析日益增长的需求，我们开发并验证了一种用于定量分析14种内源性大麻素和同系物的测定方法。采用乙腈沉淀蛋白质的简单提取，然后在线捕获高效液相色谱-串联质谱法（LC/LC-MS/MS）用于监测血浆中14种内源性大麻素的水平。对该测定方法进行了验证，并测定了运行内和运行间准确度和不精密度以及基质效应、回收率和样品稳定性。作为概念证明，分析了自然施用大麻花和浓缩物后的研究样品的子集。除N-油酰多巴胺和油酰胺外，所有内源性大麻素均符合预定验收标准。回收率可重现，未观察到显著的基质效应。样品稳定性是一个问题。对概念验证研究样本的分析显示，与鲜花使用者（252 ± 11 pg/mL）相比，浓缩物使用者（300 ± 13 pg/mL）中的二十二碳四烯酰乙醇酰胺浓度显著（p= 0.006）更高。一个强大的，灵敏的高通量测定14内源性大麻素和同系物的定量成功地验证。我们的研究表明，必须（A）适当稳定样品和（B）分离并分别定量1-AG和2-AG;否则，研究结果不可靠。对大麻花使用者和大麻浓缩物使用者的研究样本进行分析，发现二十二碳四烯酰乙醇酰胺和大麻素（n.s.）在高THC浓度的用户根据现有的文献，支持测定测量的有效性。图形摘要
The endocannabinoid system (ECS) is a complex cell-signaling system. To address the growing need of analytics capturing endocannabinoid levels to investigate the ECS, we developed and validated an assay for the quantitative analysis of 14 endocannabinoids and congeners. A simple extraction using protein precipitation with acetonitrile followed by online-trapping high-performance liquid chromatography–tandem mass spectrometry (LC/LC-MS/MS) was used to monitor the levels of 14 endocannabinoids in plasma. The assay was validated and intra-run and inter-run accuracies and imprecisions as well as matrix effects, recoveries, and sample stabilities were determined. As a proof of concept, a subset of study samples after naturalistic administration ofCannabisflower and concentrate was analyzed. With the exception of N-oleoyl dopamine and oleamide, all endocannabinoids fulfilled the predefined acceptance criteria. Reproducible recoveries and no significant matrix effects were observed. Sample stability was an issue. Analysis of the proof-of-concept study samples revealed a significantly (p= 0.006) higher concentration of docosatetraenoyl ethanolamide in concentrate users (300 ± 13 pg/mL) compared to flower users (252 ± 11 pg/mL). A robust, sensitive high-throughput assay for the quantitation of 14 endocannabinoids and congeners was successfully validated. Our study showed that it is mandatory to (A) appropriately stabilize samples and (B) separate and separately quantify 1-AG and 2-AG; otherwise, study results are unreliable. The analysis of study samples fromCannabisflower users versusCannabisconcentrate users revealed higher levels of docosatetraenoyl ethanolamide and anandamide (n.s.) in high THC concentrate users in accordance with the existing literature, supporting the validity of the assay measurements.Graphical abstract