Moon Ju-Yeon, Kim Jin Young, Moon Myeong Hee, Chung Bong Chul, In Moon Kyo, Choi Man Ho
Life Sciences Division, Korea Institute of Science & Technology, 39-1 Hawolkok-dong, Seoul 136-791, South Korea.
J Chromatogr A. 2008 Sep 12;1204(1):87-92. doi: 10.1016/j.chroma.2008.07.083. Epub 2008 Aug 5.
A comprehensive solid-phase extraction (SPE) technique based on the formation of an inclusion complex between beta-cyclodextrin (betaCD) and cannabinoids including Delta(9)-tetrahydrocannabinol (THC), 11-hydroxy-Delta(9)-tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol (THC-COOH) was developed in gas chromatographic-mass spectrometric (GC-MS) analysis. A betaCD/epichlorohydrin copolymer was prepared and then 'hardened' in aqueous solution with 0.3M CaCl(2) to yield a stable particulate copolymer, which was used as a novel SPE sorbent. An internal standard THC-COOH-d(9) was added to urine samples containing 3 cannabinoids and then purified with the hardened betaCD polymer. The cannabinoids were extracted from the hardened betaCD using tetrahydrofuran. Resulting extracts were evaporated and derivatized with MSTFA/NH(4)I/dithioerythritol (500:4:2, v/w/w) and analyzed by GC-MS in selected-ion monitoring (SIM) mode. Overall recoveries ranged from 85% to 102%, with a detection limit of 0.2 microg L(-1) for the three cannabinoids tested. The precision (% CV) and accuracy (% bias) of the assay were 1.2-5.1% and 93-111% in 0.2-50 microg L(-1) calibration range, respectively (r(2)>0.9997). Twenty actual samples positive by fluorescence polarization immunoassay were also quantitatively analyzed. The devised technique based on the calcium-hardened betaCD sorption of cannabinoids and subsequent GC-SIM/MS resulted in better selectivity and extraction efficiency than is possible using the conventional hydrophobicity-based SPE methods.
开发了一种基于β-环糊精（βCD）与大麻素（包括Δ⁹-四氢大麻酚（THC）、11-羟基-Δ⁹-四氢大麻酚（11-OH-THC）和11-去甲-9-羧基-Δ⁹-四氢大麻酚（THC-COOH））形成包合物的综合固相萃取（SPE）技术，用于气相色谱-质谱（GC-MS）分析。制备了βCD/环氧氯丙烷共聚物，然后在0.3M氯化钙水溶液中“硬化”，得到稳定的颗粒状共聚物，用作新型SPE吸附剂。将内标THC-COOH-d⁹添加到含有3种大麻素的尿液样本中，然后用硬化的βCD聚合物进行纯化。使用四氢呋喃从硬化的βCD中提取大麻素。将所得提取物蒸发，并用MSTFA/NH₄I/二硫赤藓糖醇（500:4:2，v/w/w）进行衍生化，然后在选择离子监测（SIM）模式下通过GC-MS进行分析。总体回收率在85%至102%之间，所测试的三种大麻素的检测限为0.2μg L⁻¹。在0.2至50μg L⁻¹的校准范围内，该测定的精密度（%CV）和准确度（%偏差）分别为1.2 - 5.1%和93 - 111%（r²>0.9997）。还对20份荧光偏振免疫测定呈阳性的实际样本进行了定量分析。基于钙硬化βCD吸附大麻素并随后进行GC-SIM/MS的设计技术，比使用传统的基于疏水性的SPE方法具有更好的选择性和提取效率。
