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. 2013 Jun;5(12):1501-6.
doi: 10.4155/bio.13.91.

Determination of artemether and dihydroartemisinin in human plasma with a new hydrogen peroxide stabilization method

Liusheng Huang  1 , Alexander Olson, David Gingrich, Francesca T Aweeka
Affiliations

Determination of artemether and dihydroartemisinin in human plasma with a new hydrogen peroxide stabilization method

Liusheng Huang et al. Bioanalysis. 2013 Jun.

Abstract

Background: Numerous methods have been reported for the determination of artemether (ARM) and its metabolite dihydroartemisinin (DHA) in plasma. However, stability issues in patient plasma have not received enough attention.

Results: An LC-MS/MS method for simultaneous determination of ARM and DHA in human plasma (K3EDTA) turned out to be problematic: ARM and DHA were degraded partially or completely in some patient plasma samples as indicated by the stable isotope-labeled internal standards. We postulated iron II (Fe(2+)) in hemoglobin or its derived products from malaria patients causes degradation of the drugs, and found that hydrogen peroxide (H2O2) protected the drugs from degradation. Acidifying plasma increased recovery of ARM significantly. Using only 50 µl of plasma sample, the method has a LLOQ at 0.5 ng/ml for both ARM and DHA.

Conclusion: H2O2 is a stabilizing agent for artemisinin derivatives. The modified method is reliable and sensitive.

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Conflict of interest statement

Financial & competing interests disclosure

The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.

Figures

Figure 1
Figure 1. Artemether, dihydroartemisinin and their corresponding internal standards
Figure 2
Figure 2. Chromatograms of blank plasma (light line) and samples at LLOQ level (dark line)
DHA: Dihydroartemisinin.
Figure 3
Figure 3. Comparison of signal intensity of internal standard for patient samples when added in three different solutions
(A) Artemether-13CD3 and (B) dihydrosrtemisinin-d3. IS: Internal standard.
See this image and copyright information in PMC

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