Stanley Scott D, McKemie Daniel, Skinner Wayne
University of California, Davis, Kenneth L Maddy Equine Analytical Chemistry Laboratory, West Health Science Drive, Davis, California 95616, USA.
J Anal Toxicol. 2003 Sep;27(6):325-31. doi: 10.1093/jat/27.6.325.
A rapid, sensitive, and rugged method for detecting drugs and drug metabolites in extracts of horse urine is described. The use of large-volume injection (LVI) gas chromatography-mass spectrometry (GC-MS) for analysis of horse urine extracts allowed automation of the derivatization procedure and reduction of the sample volume from 5 mL to 1 mL of urine. An autosampler and temperature-programmable inlet were used to automatically dissolve the sample extract and form trimethylsilyl derivatives of over 200 analytes. The suitability of this procedure for routine GC-MS detection of approximately 80 basic analytes in extracts of racehorse urine was investigated. The formation of derivatives using LVI with in-liner derivatization was compared to a manual procedure involving the dissolution of sample extracts in N,O-bis(trimethylsily)trifluoroacetamide, heating the resulting mixture, and injecting 1 or 2 microL of the mixture through a splitless injector into the GC-MS instrument. In all cases, the in-liner derivatization reactions were found to be as complete as conventional heating block procedures. Ruggedness testing of the method demonstrated that peak resolution, shape, and area were maintained through 40 consecutive injections of sample extracts. No evidence of the accumulation of interfering substances was observed. The limits of detection using LVI GC-MS for routine screening of basic drugs in urine were generally in the range of 5-25 ng/mL. The method is currently being used to detect basic analytes in horse urine extracts with a throughput of approximately 50 urine sample extracts per instrument per day.
本文描述了一种快速、灵敏且耐用的方法，用于检测马尿提取物中的药物及其代谢物。使用大体积进样（LVI）气相色谱-质谱联用（GC-MS）分析马尿提取物，可实现衍生化过程的自动化，并将样品体积从5 mL尿液减少至1 mL。使用自动进样器和温度可编程进样口自动溶解样品提取物，并形成200多种分析物的三甲基硅烷基衍生物。研究了该方法用于赛马尿提取物中约80种碱性分析物常规GC-MS检测的适用性。将使用LVI在线衍生化形成衍生物的方法与手动方法进行了比较，手动方法包括将样品提取物溶解在N,O-双（三甲基硅基）三氟乙酰胺中，加热所得混合物，然后通过不分流进样器将1或2 μL混合物注入GC-MS仪器。在所有情况下，发现在线衍生化反应与传统加热块程序一样完全。该方法的耐用性测试表明，通过连续40次进样样品提取物，峰分辨率、峰形和峰面积得以保持。未观察到干扰物质积累的迹象。使用LVI GC-MS对尿液中的碱性药物进行常规筛查的检测限通常在5-25 ng/mL范围内。该方法目前正用于检测马尿提取物中的碱性分析物，每台仪器每天的通量约为50个尿样提取物。
