PDF(Pubmed)
Abstract：
Recently, hexahydrocannabinol (HHC) was posed under strict control in Europe due to the increasing HHC-containing material seizures. The lack of analytical methods in clinical laboratories to detect HHC and its metabolites in biological matrices may result in related intoxication underreporting. We developed and validated a comprehensive GC-MS/MS method to quantify 9(R)-HHC, 9(S)-HHC, 9αOH-HHC, 9βOH-HHC, 8(R)OH-9(R)-HHC, 8(S)OH-9(S)HHC, 11OH-9(R)HHC, 11OH-9(S)HHC, 11nor-carboxy-9(R)-HHC, and 11nor-carboxy-9(S)-HHC in whole blood, urine, and oral fluid. A novel QuEChERS extraction protocol was optimized selecting the best extraction conditions suitable for all the three matrices. Urine and blood were incubated with β-glucuronidase at 60 °C for 2 h. QuEChERS extraction was developed assessing different ratios of Na2SO4:NaCl (4:1, 2:1, 1:1, w/w) to be added to 200 µL of any matrix added with acetonitrile. The chromatographic separation was achieved on a 7890B GC with an HP-5ms column, (30 m, 0.25 mm × 0.25 µm) in 12.50 min. The analytes were detected with a triple-quadrupole mass spectrometer in the MRM mode. The method was fully validated following OSAC guidelines. The method showed good validation parameters in all the matrices. The method was applied to ten real samples of whole blood (n = 4), urine (n = 3), and oral fluid (n = 3). 9(R)-HHC was the prevalent epimer in all the samples (9(R)/9(S) = 2.26). As reported, hydroxylated metabolites are proposed as urinary biomarkers, while carboxylated metabolites are hematic biomarkers. Furthermore, 8(R)OH-9(R)HHC was confirmed as the most abundant metabolite in all urine samples.
摘要：
最近,六氢大麻酚（HHC）在欧洲受到严格控制，因为含有HHC的物质缉获量不断增加。临床实验室缺乏检测生物基质中HHC及其代谢物的分析方法可能导致相关的中毒漏报。我们开发并验证了一种全面的GC-MS/MS方法来量化9(R)-HHC，9(S)-HHC,9αOH-HHC,9βOH-HHC,8(R)OH-9(R)-HHC,8(S)OH-9(S)HHC,11OH-9(R)HHC,11OH-9(S)HHC,11nor-羧基-9（R）-HHC，和全血中的11nor-羧基-9(S)-HHC，尿液,和口服液。优化了一种新型QuEChERS提取方案，选择了适合所有三种基质的最佳提取条件。将尿液和血液与β-葡糖醛酸糖苷酶在60°C下孵育2小时。开发QuEChERS提取，评估要添加到200μL添加有乙腈的任何基质中的不同比例的Na2SO4：NaCl（4：1、2：1、1：1，w/w）。用HP-5ms色谱柱在7890BGC上实现色谱分离，(30米,0.25mm×0.25µm）在12.50分钟内。用三重四极杆质谱仪以MRM模式检测分析物。该方法按照OSAC指南进行了充分验证。该方法在所有矩阵中都显示出良好的验证参数。该方法应用于10份全血样本(n=4)，尿液（n=3），和口服液（n=3）。9（R）-HHC是所有样品中普遍存在的差向异构体（9（R）/9（S）=2.26）。据报道，羟基化代谢物被提议作为尿液生物标志物，而羧化代谢物是血液生物标志物。此外，证实8(R)OH-9(R)HHC是所有尿样中最丰富的代谢物。
