Salamone S J, Honasoge S, Brenner C, McNally A J, Passarelli J, Goc-Szkutnicka K, Brenneisen R, elSohly M A, Feng S
Roche Diagnostic Systems, Somerville, New Jersey 08876, USA.
J Anal Toxicol. 1997 Sep;21(5):341-5. doi: 10.1093/jat/21.5.341.
A study was conducted to compare the performance of the OnLine and OnTrak immunoassays for benzodiazepines with gas chromatographic-mass spectrometric (GC-MS) analysis in detecting flunitrazepam (FNP) and its metabolites in human urine. Urine was collected over a 72-h period from six individuals (four male and two female) who had taken a single oral dose of either 1 or 4 mg of FNP. The OnTrak assay was run at a 100-ng/mL cutoff of nordiazepam (NDP), and the OnLine assay was run with a standard curve from zero to 200 ng/mL of NDP with and without beta-glucuronidase treatment. Each sample was analyzed by GC-MS using FNP, 7-amino-FNP, 3-hydroxy-FNP, desmethyl-FNP, 7-amino-3-hydroxy-FNP, and desmethyl-3-hydroxy-FNP as standards with beta-glucuronidase treatment. The specimens from the 1-mg dose did not yield a positive result by immunoassay over the 72-h collection period. Specimens from the 4-mg dose did yield positive results in both immunoassays. The time of the first positive result ranged from 4 to 12 h, and the time to the last positive result ranged from 18 to 60 h. Treatment of the samples with beta-glucuronidase increased the OnLine values between 20 and 60%, but it did not appreciably increase the detection time. GC-MS analysis showed no detectable levels of FNP, 3-hydroxy-FNP, desmethyl-FNP, 7-amino-3-hydroxy-FNP, and desmethyl-3-hydroxy-FNP. However, all samples collected past time zero showed detectable levels of 7-amino-FNP (> 2 ng/mL) with peak concentrations at 12-36 h. The peak levels of 7-amino-FNP by GC-MS paralleled the peak levels of the immunoassay response. The amount of 7-amino-FNP metabolite quantitated by GC-MS, however, accounted for only 15-20% of the total immunoassay crossreactive FNP metabolites.
开展了一项研究，比较在线免疫分析和OnTrak免疫分析检测苯二氮卓类药物与气相色谱 - 质谱联用（GC - MS）分析法在检测人尿中氟硝西泮（FNP）及其代谢物方面的性能。在72小时内从6名个体（4名男性和2名女性）收集尿液，这些个体单次口服1或4毫克FNP。OnTrak分析以去甲西泮（NDP）100纳克/毫升的临界值进行，在线分析使用从零到200纳克/毫升NDP的标准曲线，有或没有β - 葡萄糖醛酸酶处理。每个样品通过GC - MS分析，使用FNP、7 - 氨基 - FNP、3 - 羟基 - FNP、去甲基 - FNP、7 - 氨基 - 3 - 羟基 - FNP和去甲基 - 3 - 羟基 - FNP作为标准品并进行β - 葡萄糖醛酸酶处理。来自1毫克剂量的样本在72小时收集期内免疫分析未产生阳性结果。来自4毫克剂量的样本在两种免疫分析中均产生阳性结果。首次阳性结果的时间范围为4至12小时，最后阳性结果的时间范围为18至60小时。用β - 葡萄糖醛酸酶处理样品使在线分析值增加了20%至60%，但并未明显增加检测时间。GC - MS分析显示未检测到FNP、3 - 羟基 - FNP、去甲基 - FNP、7 - 氨基 - 3 - 羟基 - FNP和去甲基 - 3 - 羟基 - FNP的水平。然而，所有零时之后收集的样品均显示可检测到的7 - 氨基 - FNP水平（>2纳克/毫升），在12至36小时达到峰值浓度。GC - MS检测的7 - 氨基 - FNP峰值水平与免疫分析响应的峰值水平平行。然而，通过GC - MS定量的7 - 氨基 - FNP代谢物量仅占免疫分析交叉反应性FNP代谢物总量的15%至20%。
