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

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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.