Plecko Thomas, Berbalk Kevin, Wieland Eberhard, Shipkova Maria
Zentralinstitut für Klinische Chemie und Laboratoriumsmedizin, Klinikum Stuttgart, Stuttgart, Germany.
Ther Drug Monit. 2018 Oct;40(5):642-648. doi: 10.1097/FTD.0000000000000544.
Oral fluid (OF) is increasingly used as an alternative sample matrix in drug of abuse screening. Screening is commonly performed by immunoassays and results confirmed using laborious gas chromatography-mass spectrometry (GC-MS)-based methods. Therefore, an easy to operate ion trap mass spectrometric (IT-MS) commercial screening method (Toxtyper; Bruker Daltronik, Bremen, Germany) combined with a laboratory-developed sample preparation procedure has been evaluated for their application to OF.
OF samples were subjected to protein precipitation followed by HybridSPE-Phospholipid extraction. Chromatographic separation was achieved by ultra-high-performance liquid chromatography; MS2/MS3 spectra were recorded by IT-MS and analyzed using a library provided by the manufacturer (Bruker Daltronik). The lower limit of detection, linearity, imprecision, inaccuracy, and specificity (interferences and matrix effects) were investigated for methadone, buprenorphine, pregabalin, fentanyl, amphetamine, 3,4-methylendioxy-N-methylamphetamine, cocaine, acetylcodeine, and nordiazepam, after spiking drug-free OF with these test substances. In addition, concordance between IT-MS results and gas chromatography-tandem mass spectrometry, liquid chromatography-tandem mass spectrometry, or immunoassay (buprenorphine) results was investigated.
No interferences or matrix effects were observed. The lower limit of detection for acetylcodeine, amphetamine, benzoylecgonine, methadone, and nordiazepam was below the common cutoffs for immunological screening assays and comparable to that of GC-MS. Imprecision and inaccuracy, both in- and between-series, were consistently <25%, except for buprenorphine. Toxtyper screening for pregabalin and fentanyl was less sensitive than a targeted liquid chromatography-tandem mass spectrometry assay. A very good concordance was found between the previous analytical approach and the new IT-MS method.
The Toxtyper IT-MS is easy to use and can be applied for the screening of drug of abuse and the qualitative confirmation analysis in OF in a clinical toxicology service. Although intended for qualitative analysis, performance data suggest that the methods investigated may also be applicable for semiquantitative longitudinal follow-up.
口腔液（OF）越来越多地被用作滥用药物筛查的替代样本基质。筛查通常通过免疫测定法进行，结果使用基于繁琐的气相色谱 - 质谱联用（GC - MS）的方法进行确认。因此，已经评估了一种易于操作的离子阱质谱（IT - MS）商业筛查方法（Toxtyper；德国不来梅的布鲁克道尔顿公司）与实验室开发的样本制备程序在OF中的应用。
OF样本先进行蛋白质沉淀，然后进行HybridSPE - 磷脂提取。通过超高效液相色谱实现色谱分离；IT - MS记录MS2/MS3光谱并使用制造商（布鲁克道尔顿公司）提供的库进行分析。在无药物的OF中加入这些测试物质后，研究了美沙酮、丁丙诺啡、普瑞巴林、芬太尼、苯丙胺、3,4 - 亚甲基二氧基 - N - 甲基苯丙胺、可卡因、乙酰可待因和去甲西泮的检测下限、线性、不精密度、不准确和特异性（干扰和基质效应）。此外，还研究了IT - MS结果与气相色谱 - 串联质谱、液相色谱 - 串联质谱或免疫测定法（丁丙诺啡）结果之间的一致性。
未观察到干扰或基质效应。乙酰可待因、苯丙胺、苯甲酰爱康宁、美沙酮和去甲西泮的检测下限低于免疫筛查测定的常见临界值，且与GC - MS相当。除丁丙诺啡外，批内和批间的不精密度和不准确均始终<25%。Toxtyper对普瑞巴林和芬太尼的筛查比靶向液相色谱 - 串联质谱测定法灵敏度低。发现先前的分析方法与新的IT - MS方法之间具有非常好的一致性。
Toxtyper IT - MS易于使用，可应用于临床毒理学服务中OF的滥用药物筛查和定性确证分析。尽管旨在进行定性分析，但性能数据表明所研究的方法也可能适用于半定量纵向随访。
