Hayakawa K, Hasegawa K, Imaizumi N, Wong O S, Miyazaki M
Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
J Chromatogr. 1989 Mar 3;464(2):343-52. doi: 10.1016/s0021-9673(00)94252-3.
High-performance liquid chromatography with chemiluminescence detection has been established for the determination of trace levels of amphetamine-related compounds (APs) after fluorigenic derivatization. Bis(2,4,6-trichlorophenyl) oxalate and hydrogen peroxide in acetonitrile was used as a post-column chemilumigenic reagent. As derivatization reagents, dansyl chloride (Dns-Cl), 4-fluoro-7-nitrobenzoxadiazole (NBD-F) and naphtalene-2,3-dicarbaldehyde (NDA) were compared. Dns-Cl was the most suitable of the three for the simultaneous determination of both primary and secondary amino APs. The on-column detection limits for Dns-derivatives were 3.10(-15) - 4.10(-15) mol. NDA gave the most sensitive derivatives (cyanobenz[f]isoindole, CBI derivatives) with only primary amino APs. The detection limits for CBI derivatives were as low as 2.10(-16) mol. The present method was applied to the determination of methamphetamine in human urine. Only diethyl ether extraction was necessary as a clean-up treatment before Dns derivatization, because diethyl ether extracted methamphetamine quantitatively from urine at strong alkaline pH and the extract showed few interfering peaks around the retention time of methamphetamine after the derivatization. Methamphetamine concentrations as low as 1.10(-7) M in urine were determined after the above treatments.
建立了一种高效液相色谱-化学发光检测法，用于测定荧光衍生化后痕量的苯丙胺类相关化合物（APs）。以乙腈中的双（2,4,6-三氯苯基）草酸酯和过氧化氢作为柱后化学发光试剂。比较了丹磺酰氯（Dns-Cl）、4-氟-7-硝基苯并恶二唑（NBD-F）和萘-2,3-二甲醛（NDA）作为衍生化试剂。在同时测定伯胺和仲胺类APs方面，Dns-Cl是三者中最合适的。Dns衍生物的柱上检测限为3×10⁻¹⁵ - 4×10⁻¹⁵ mol。NDA仅对伯胺类APs产生最灵敏的衍生物（氰基苯并[f]异吲哚，CBI衍生物）。CBI衍生物的检测限低至2×10⁻¹⁶ mol。该方法应用于人体尿液中甲基苯丙胺的测定。在进行Dns衍生化之前，仅需用乙醚萃取作为净化处理，因为在强碱性pH条件下，乙醚能从尿液中定量萃取甲基苯丙胺，且衍生化后萃取液在甲基苯丙胺保留时间附近几乎没有干扰峰。经过上述处理后，可测定尿液中低至1×10⁻⁷ M的甲基苯丙胺浓度。
