I want to quantify artemisinin content of plant samples through HPLC, but some how I'm unable to validate it. As per the papers I'm using phosphate buffer: methanol (40:60) pH 6.4 as mobile phase. Also dissolved the standard artemisinin (sigma) in methanol (1mg/ml)+ 0.2% NaOH (4ml), incubated at 50 degree for 30 mins, then cooling it to room temperature and again adding 0.1M acetic acid (5ml). RP C18 and wavelength @ 260nm. no peaks visible at this particular wavelength.
The monographs of World Health Organization (WHO) describe an HPLC method without derivatization using UV detection at 210nm. Many authors discarded the WHO HPLC method on the basis of very low UV absorbance of artemisinin.
There is an excellent publication out of Rachel Gomes' group describing a straightforward HPLC-UV analysis of artemisinin in plant samples. It's in the Journal of Pharmaceutical & Biomedical Analysis, volume 70, 136.
You can also try the method described in this paper, using 192 nm: http://www.ncbi.nlm.nih.gov/pubmed/18980258
It seems pretty straight forward. I would also be careful about the solvent used in the sample preparation, since using a solvent that is too different (polarity) to your starting eluent might result in poor peak shape and deviations in RT.
Good luck
Hi,
You can try simultaneous UV detection at 192, 210 and 260 nm. After this you may be get some idea about peak intensity. Most of researchers reported 210 nm wavelength is working for Artemisinin. Good luck.
We analysed plants after extraction with methanol using LC-MS/MS. Verry easy and sensitiv. No problems with low wavelength (210 nm) and bachground. If you need more information on method, let me know.
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