Lindegardh Niklas, Hanpithakpong Warunee, Kamanikom Benjamas, Pattayaso Janhom, Singhasivanon Pratap, White Nicholas J, Day Nicholas P J
Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Bioanalysis. 2011 Jul;3(14):1613-24. doi: 10.4155/bio.11.158.
Quantification of artemisinin (ARN) and its derivatives in whole blood has hitherto been thought impossible.
A LC-MS/MS method for the analysis of artesunate (ARS), its metabolite dihydroartemisinin (DHA) and artemisinin in human whole blood has been developed and successfully validated. The method includes stabilization of the blood matrix at the time of collection and at the time of analysis. Addition of potassium dichromate to the blood samples deactivated the Fe(2+) core in hemoglobin, while deferoxamine chelated Fe(3+) and prevented back conversion into Fe(2+). A pilot study showed that the blood:plasma ratio for ARS and DHA is approximately 0.75, indicating a significantly lower uptake in red blood cells than had previously been estimated using radiolabeled drug methodology.
The developed LC-MS/MS assay is the first method available for quantification of ARN and its derivatives in blood and opens up new possibilities of studying these drugs inside infected red blood cells.
迄今为止，人们一直认为无法对全血中的青蒿素（ARN）及其衍生物进行定量分析。
已开发出一种用于分析人全血中青蒿琥酯（ARS）、其代谢物双氢青蒿素（DHA）和青蒿素的液相色谱-串联质谱（LC-MS/MS）方法，并成功进行了验证。该方法包括在采集血液样本时和分析时对血液基质进行稳定处理。向血液样本中添加重铬酸钾可使血红蛋白中的Fe(2+)核心失活，而去铁胺可螯合Fe(3+)并防止其逆转为Fe(2+)。一项初步研究表明，ARS和DHA的血-浆比约为0.75，这表明红细胞对其摄取量明显低于先前使用放射性标记药物方法所估计的量。
所开发的LC-MS/MS测定法是第一种可用于定量血液中ARN及其衍生物的方法，为研究这些药物在受感染红细胞内的情况开辟了新的可能性。
