Chemistry and Drug Metabolism, National Institute on Drug Abuse, National Institutes of Health, 251 Bayview Blvd. Suite 200, Room 05A721A, Baltimore, MD, USA.
Anal Bioanal Chem. 2010 Jul;397(6):2335-47. doi: 10.1007/s00216-010-3772-y. Epub 2010 Jun 2.
Identification of prenatal cannabis exposure is important due to potential cognitive and behavioral consequences. A two-dimensional gas chromatography-mass spectrometry method for cannabinol, Delta(9)-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), 8beta,11-dihydroxy-THC, and 11-nor-9-carboxy-THC (THCCOOH) quantification in human meconium was developed and validated. Alkaline, enzymatic, and enzyme-alkaline tandem hydrolysis conditions were optimized with THC- and THCCOOH-glucuronide reference standards. Limits of quantification ranged from 10 to 15 ng/g, and calibration curves were linear to 500 ng/g. Bias and intra-day and inter-day imprecision were <12.3%. Hydrolysis efficiencies were analyte-dependent; THC-glucuronide was effectively cleaved by enzyme, but not base. Conversely, THCCOOH-glucuronide was most sensitive to alkaline hydrolysis. Enzyme-alkaline tandem hydrolysis maximized efficiency for both glucuronides. Identification of cannabinoid-positive meconium specimens nearly doubled following alkaline and enzyme-alkaline hydrolysis. Although no 11-OH-THC glucuronide standard is available, enzymatic hydrolysis improved 11-OH-THC detection in authentic specimens. Maximal identification of cannabis-exposed neonates and the widest range of cannabis biomarkers are achieved with enzyme-alkaline tandem hydrolysis.
由于潜在的认知和行为后果，识别产前大麻暴露非常重要。本文建立并验证了一种二维气相色谱-质谱法，用于检测人胎粪中大麻酚、Delta(9)-四氢大麻酚（THC）、11-羟基-THC（11-OH-THC）、8beta,11-二羟基-THC 和 11-去甲-9-羧酸-THC（THCCOOH）。采用 THC 和 THCCOOH-葡萄糖醛酸苷参考标准对碱性、酶解和酶-碱串联水解条件进行了优化。定量下限范围为 10-15ng/g，校准曲线在 500ng/g 范围内呈线性。偏差、日内和日间精密度均<12.3%。水解效率取决于分析物；THC-葡萄糖醛酸苷可被酶有效裂解，但不能被碱裂解。相反，THCCOOH-葡萄糖醛酸苷对碱水解最敏感。酶-碱串联水解使两种葡萄糖醛酸苷的水解效率最大化。经碱和酶-碱水解后，大麻阳性胎粪标本的鉴定率几乎翻了一番。尽管没有 11-OH-THC-葡萄糖醛酸苷标准品，但酶解可提高真实标本中 11-OH-THC 的检测率。采用酶-碱串联水解可最大程度地鉴定出大麻暴露的新生儿，并检测到最广泛的大麻生物标志物。
