Kumihashi Mitsuru, Ameno Kiyoshi, Shibayama Takayuki, Suga Keisuke, Miyauchi Hiroshi, Jamal Mostofa, Wang Weihuan, Uekita Ikuo, Ijiri Iwao
Department of Forensic Medicine, Faculty of Medicine, Kagawa University, 1750-1 Ikenobe, Miki, Kita, Kagawa 761-0793, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jan 1;845(1):180-3. doi: 10.1016/j.jchromb.2006.07.049. Epub 2006 Aug 17.
We describe here a simple, precise, and highly sensitive method for the simultaneous determination of methamphetamine (MA) and amphetamine (AM) in urine using a high performance liquid chromatography (HPLC) column-switching method. A PK-2A (Shodex) column was used for extraction and deproteinization, and a CAPCELL PAK SCX semi-micro, polymer-coated cation-exchange column was employed for separation. The urine sample was mixed with an equal volume of borate buffer (0.1M, pH 9.4), and then 100 microl of the mixture was injected into the HPLC column. The column was switched for 6 min, and then 10 min later detection was performed at 210 nm. Recovery yields of the MA and AM spiked in the urine were 93.0-100.4% with a coefficient of variation of less than 1%. The calibration curves of MA and AM were in the range of 0.1-10 microg/ml with good linearity (r(2)=0.999), with the limit of qualification being 0.005 microg/ml. This method of using HPLC with column-switching can be used for both qualification and quantification of MA and its metabolite, AM, in urine, especially in forensic cases.
我们在此描述一种使用高效液相色谱（HPLC）柱切换法同时测定尿液中甲基苯丙胺（MA）和苯丙胺（AM）的简单、精确且高度灵敏的方法。采用PK - 2A（昭和电工）柱进行萃取和脱蛋白，使用CAPCELL PAK SCX半微聚合物涂层阳离子交换柱进行分离。将尿液样本与等体积的硼酸盐缓冲液（0.1M，pH 9.4）混合，然后将100微升该混合物注入HPLC柱。柱切换6分钟，然后10分钟后在210纳米处进行检测。尿液中添加的MA和AM的回收率为93.0 - 100.4%，变异系数小于1%。MA和AM的校准曲线在0.1 - 10微克/毫升范围内，线性良好（r² = 0.999），定量限为0.005微克/毫升。这种使用HPLC柱切换的方法可用于尿液中MA及其代谢物AM的定性和定量分析，特别是在法医案件中。
