Development and Application of chiral separation HPLC methods with fluorescence or chemiluminescence detection for determination of metamphetamines

以新合成的荧光试剂4-(4，5-二苯基-1H-咪唑-2-基)苯甲酰氯(DIB-Cl)为标记物，建立了测定甲基苯丙胺(MPS)的高效液相色谱方法。采用过草酸化学发光法(PO-CL)和荧光法对甲基苯丙胺、苯丙胺、对羟基甲基苯丙胺和麻黄碱等MPS的DIB衍生物进行了分离和检测。由于荧光检测法比PO-CL法更灵敏，因此本研究选择了高效液相色谱-荧光检测法。建立的方法可以检测到阿摩尔~f摩尔水平的甲基苯丙胺。用该方法成功地测定了部分吸毒者的尿液和头发样本。建立了手性和手性半微柱高效液相色谱方法，可用于10μL尿样和1厘米长毛发样品中MPS的测定，方法简便、灵敏，可用于药理学、毒理学和药学研究，特别是对含有痕量MPS的样品的研究具有重要意义。

A highly sensitive HPLC method for the determination of methamphetamines (MPs) has been developed using a newly synthesized fluorescent reagent, 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. DIB derivatives of MPs, e.g., metamphetamine, amphetamine, p-hydroxymethamphetamine, and ephedrine, were separated and detected with peroxyoxalate chemiluminescence (PO-CL) and fluorescence methods. Fluorescence detection was more sensitive than PO-CL, thus HPLC with fluorescence detection was selected in this study. The developed method could detect methamphetamines at amol〜fmol levels. Some abusers' urine and hair samples were successfully determined by the method. We also focused ourselves to develop simple and economic methods by downsizing the system using semi-micro column HPLC.Achiral and chiral semi-micro column HPLC methods have been developed, which could be successfully applied to the determination of MPs in only 10 μL of urine or 1 cm-long hair samples.The proposed methods are simple and sensitive, and thus, they would be useful for pharmacological, toxicological and pharmaceutical studies, especially for the studies with samples including trace amounts of MPs.