Nakashima Kenichiro, Kaddoumi Amal, Ishida Yukihiro, Itoh Takachika, Taki Kenji
Department of Clinical Pharmacy, Course of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Biomed Chromatogr. 2003 Oct;17(7):471-6. doi: 10.1002/bmc.278.
This paper describes highly sensitive HPLC methods for the determination of amphetamine (AP) and methamphetamine (MP) in abusers' plasma and hair samples. AP and MP were derivatized with the fluorescent reagent, DIB-Cl, to yield a highly fluorescent DIB-derivatives of AP and MP, which were then analyzed by HPLC with fluorescence detection at excitation and emission wavelengths of 325 and 430 nm, respectively. The separation was achieved on an ODS column with isocratic mobile phases composed of acetoniltrile and citrate buffer (55:45, v/v) for plasma samples and of acetonitrile-methanol-citrate buffer (45:20:37.5, v/v/v) for hair samples. The limits of detection were less than 0.87 ng/mL and 0.12 ng/mg in plasma and hair samples, respectively, for both AP and MP. The methods were then applied to the determination of MP and its metabolite AP in plasma obtained from two cases of illegally ingested MP and in one of the cases' hair received later. Case I was treated with dialysis; samples before and after dialysis were analyzed by the described method. After dialysis for 5 h, the total plasma levels of AP and MP decreased from 720 to 190 ng/mL. For case II, MP and AP levels were monitored for 3 days after digestion. Total plasma levels decreased from 57 ng/mL in the day of digestion to 11 ng/mL after 3 days. In hair samples, AP and MP could also be detected in very low concentrations.
本文描述了用于测定滥用者血浆和毛发样本中苯丙胺(AP)和甲基苯丙胺(MP)的高灵敏度高效液相色谱法。AP和MP用荧光试剂DIB-Cl进行衍生化,生成AP和MP的高荧光DIB衍生物,然后通过高效液相色谱法进行分析,荧光检测的激发波长和发射波长分别为325 nm和430 nm。血浆样本在ODS柱上进行分离,等度流动相由乙腈和柠檬酸盐缓冲液(55:45,v/v)组成;毛发样本的等度流动相由乙腈 - 甲醇 - 柠檬酸盐缓冲液(45:20:37.5,v/v/v)组成。AP和MP在血浆和毛发样本中的检测限分别低于0.87 ng/mL和0.12 ng/mg。然后将这些方法应用于测定两例非法摄入MP的患者血浆中的MP及其代谢产物AP,以及其中一例患者后来提供的毛发样本。病例I接受了透析治疗;透析前后的样本采用所述方法进行分析。透析5小时后,AP和MP的血浆总水平从720 ng/mL降至190 ng/mL。对于病例II,在消化后3天内监测MP和AP水平。血浆总水平从消化当天的57 ng/mL降至3天后的11 ng/mL。在毛发样本中,也能检测到极低浓度的AP和MP。