Brenner C, Hui R, Passarelli J, Wu R, Brenneisen R, Bracher K, ElSohly M A, Ghodoussi V D, Salamone S J
Roche Diagnostic Systems, Somerville, NJ 08876, USA.
Forensic Sci Int. 1996 May 17;79(1):31-41. doi: 10.1016/0379-0738(96)01888-9.
A study was performed to compare the ONLINE and EMIT II immunoassays with gas chromatographic/mass spectrometric (GC/MS) analysis of methaqualone metabolites on urine using samples obtained from a clinical study. Urine was collected over a 72 h period from six healthy adults (4 male, 2 female) after oral dosing with 200 mg methaqualone (MTQ). Each urine sample was analyzed by ONLINE and EMIT II. The samples were then analyzed by GC/MS, hydrolyzed with beta-glucuronidase and again analyzed by GC/MS. Both immunoassays showed greater than 600 ng/ml concentrations of drug in each sample by the second void and remained highly positive for the rest of the 72 h. Unhydrolyzed samples analyzed by GC/MS showed both low concentrations of MTQ as well as its five major hydroxylated metabolites. The hydrolyzed samples analyzed by GC/MS showed high concentrations of the hydroxylated metabolites with the 2'-hydroxy and 3'-hydroxy metabolites being present at the highest concentrations, the 4'-hydroxy metabolite at a lower amount and the 6-hydroxy and 2-hydroxy metabolites at the lowest concentrations. The GC/MS data coupled with the antibody cross-reactivity data indicate that the major species in clinical samples that cross-react in both immunoassays are the conjugated forms of the hydroxylated metabolites of MTQ. Therefore when confirming by GC/MS after an immunoassay screen it would be prudent to confirm for the major hydroxylated metabolites as glucuronides of MTQ instead of the parent drug.
开展了一项研究，使用从一项临床研究中获取的样本，将甲喹酮代谢物的在线免疫分析（ONLINE）和放射免疫分析（EMIT II）与气相色谱/质谱联用（GC/MS）分析进行比较。对6名健康成年人（4名男性，2名女性）口服200毫克甲喹酮（MTQ）后72小时内收集的尿液进行分析。每个尿液样本均通过ONLINE和EMIT II进行分析。然后对样本进行GC/MS分析，用β-葡萄糖醛酸苷酶水解后再次进行GC/MS分析。两种免疫分析均显示，在第二次排尿时每个样本中的药物浓度均高于600纳克/毫升，并且在72小时的剩余时间内一直呈强阳性。通过GC/MS分析的未水解样本显示MTQ及其五种主要羟基化代谢物的浓度均较低。通过GC/MS分析的水解样本显示羟基化代谢物浓度较高，其中2'-羟基和3'-羟基代谢物浓度最高，4'-羟基代谢物含量较低，6-羟基和2-羟基代谢物浓度最低。GC/MS数据与抗体交叉反应数据表明，在两种免疫分析中发生交叉反应的临床样本中的主要物质是MTQ羟基化代谢物的结合形式。因此，在免疫分析筛查后通过GC/MS进行确证时，谨慎的做法是确证MTQ主要羟基化代谢物的葡萄糖醛酸苷，而不是母体药物。
