ELSEVIER Journal of Chromatography A, 695 (1995) 237-242 JOURNALOF CHROMATOGRAPHY A Chemiluminescent detection of artemisinin Novel endoperoxide analysis using luminol without hydrogen peroxide Michael D. Green a'* , Dwight L. Mount a, G. Daniel Todd a, Anthony C. Capomacchia b aEntomology Branch, Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, US Department of Health and Human Services, 1600 Clifton Road, Mailstop F-12, Atlanta, GA 30333, USA ~Department of Pharmaceutics, College of Pharmacy, University of Georgia, Athens, GA, USA First received 3 June 1994; revised manuscript received 13 December 1994; accepted 14 December 1994 Abstract A novel method for artemisinin quantitation employing high-performance liquid chromatography (HPLC) with chemiluminescence (CL) detection in the absence of hydrogen peroxide (H:O2), is reported. After elution from the HPLC column, artemisinin is combined with an alkaline solution of hematin and luminol. The resulting CL signal is detected by use of a spectrofluorometer with the excitation lamp disabled, and is proportional to artemisinin concentration. The CL method was optimized and applied to the analysis of artemisinin in spiked human serum. CL in the absence of H202 or other known oxidizing species is remarkable since such oxidizers are usually required to produce CL from luminol under alkaline conditions. Artemisinin, a naturally occurring sesquiterpene, is one of several natural products that contain an endoperoxide functional group. Since H20 2 is not needed in the analysis, the endoperoxide moiety on artemisinin is implicated as a contributing source of superoxide radicals required for the light-producing reaction with luminol. 1. Introduction Artemisinin or qinghaosu is a naturally occur- ring sesquiterpene endoperoxide (Fig. 1) that is receiving considerable attention in the treatment of malaria because of growing resistance of Plasmodium falciparum strains to traditional quinoline-based antimalarial drugs [1,2]. Detection of artemisinin has proven to be a challenge as it does not possess any sensitive or * Corresponding author. 0021-9673/95/$09.50 ~ 1995 Elsevier Science BN. All rights reserved SSDI 0021-9673(94)01236-9 .CH3 .... "H 3 o Fig. 1. Structure of artemisinin.