Lombardero N, Casanova O, Behnke M, Eyler F D, Bertholf R L
Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville 32610.
Ann Clin Lab Sci. 1993 Sep-Oct;23(5):385-94.
Analytical methods were evaluated for measuring cocaine (CO), benzoylecgonine (BE), and ecgonine methyl ester (EME) in urine and methanolic extracts from meconium and diapers by isotope dilution gas chromatography/mass spectrometry (GC/MS). Volatile derivatives of the extracted drugs were generated before GC/MS analysis. Methanolic extracts from meconium and diapers were reconstituted in drug-free urine and treated as above. The limit of detection for the GC/MS method was calculated to be approximately 11 ng per mL. Within-run coefficients of variation (CVs) for urinary CO, BE, and EME were 5.7, 5.3, and 11.4 percent, respectively (N = 10); corresponding CVs for meconium 6.4, 10.7, and 21.9 percent (N = 8). Quantitative results were linear from 25 to 10,000 ng per g of meconium and 25 to 5,000 ng per mL of urine. Day-to-day precision varied from eight percent (CV) for BE in refrigerated or frozen urine to 34 percent for EME in refrigerated meconium. Recoveries of CO, BE, and EME from urine were 63, 19, and 42 percent, respectively; corresponding recoveries from meconium were 64, 21, and 25 percent. Cocaine and metabolites were extracted from wet but meconium-free diapers into methanol, which was evaporated before reconstituting in drug-free urine and extraction on a solid phase column. The CO, BE, and EME were detected in previously drug-free meconium after portions were deposited in a diaper which was wet with drug-positive urine. Unless precautions are taken to prevent extracorporeal contamination of meconium with urine, concentrations of CO and metabolites in meconium may be substantially augmented by contamination with urine. Analysis by GC/MS of CO and metabolites extracted from diapers provides an attractive alternative to collection of urine, which is difficult and may cause discomfort for the neonate.
采用同位素稀释气相色谱/质谱联用(GC/MS)技术,对尿液、胎粪及尿布甲醇提取物中的可卡因(CO)、苯甲酰爱康宁(BE)和芽子碱甲酯(EME)的分析方法进行了评估。在GC/MS分析前,先将提取的药物制成挥发性衍生物。胎粪和尿布的甲醇提取物用不含药物的尿液复溶后,按上述方法处理。GC/MS方法的检测限经计算约为每毫升11纳克。尿液中CO、BE和EME的批内变异系数(CV)分别为5.7%、5.3%和11.4%(N = 10);胎粪中相应的CV分别为6.4%、10.7%和21.9%(N = 8)。定量结果在每克胎粪25至10,000纳克和每毫升尿液25至5,000纳克范围内呈线性关系。日间精密度变化范围从冷藏或冷冻尿液中BE的8%(CV)到冷藏胎粪中EME的34%。尿液中CO、BE和EME的回收率分别为63%、19%和42%;胎粪中相应的回收率分别为64%、21%和25%。可卡因及其代谢物从湿润但不含胎粪的尿布中提取到甲醇中,在复溶于不含药物的尿液并在固相柱上萃取前,先将甲醇蒸发。将部分药物阳性尿液浸湿的尿布放置含有先前不含药物的胎粪后,检测到了胎粪中的CO、BE和EME。除非采取预防措施防止胎粪被尿液体外污染,否则尿液污染可能会使胎粪中CO及其代谢物的浓度大幅增加。对从尿布中提取的CO及其代谢物进行GC/MS分析,为尿液采集提供了一种有吸引力的替代方法,尿液采集困难且可能给新生儿带来不适。