Affordable and rapid HPTLC method for the simultaneous analysis of artemisinin and its metabolite artemisinic acid in Artemisia annua L.
Abstract
Artemisinin (AN) and artemisinic acid (AA), valuable phyto‐pharmaceutical molecules, are well known anti‐malarials, but their activities against diseases like cancer, schistosomiasis, HIV, hepatitis‐B and leishmaniasis are also being reported. For the simultaneous estimation of AN and AA in the callus and leaf extracts of A. annua L. plants, we embarked upon a simple, rapid, selective, reliable and fairly economical high performance thin layer chromatography (HPTLC) method. Experimental conditions such as band size, chamber saturation time, migration of solvent front and slit width were critically studied and the optimum conditions were selected. The separations were achieved using toluene–ethyl acetate, 9:1 (v/v) as mobile phase on pre‐coated silica gel plates, G 60F 254. Good resolution was achieved with Rf values of 0.35 ± 0.02 and 0.26 ± 0.02 at 536 nm for AN and 626 nm for AA, respectively, in absorption–reflectance mode. The method displayed a linear relationship with r2 value 0.992 and 0.994 for AN and AA, respectively, in the concentration range of 300–1500 ng for AN and 200–1000 ng for AA. The method was validated for specificity by obtaining in‐situ UV overlay spectra and sensitivity by estimating limit of detection (30 ng for AN and 15 ng for AA) and limit of quantitation (80 ng for AN and 45 ng for AA) values. The accuracy was checked by the recovery studies conducted at three different levels with the known concentrations and the average percentage recovery was 101.99% for AN and 103.84% for AA. The precision was analyzed by interday and intraday precision and was 1.09 and 1.00% RSD for AN and 1.22 and 6.05% RSD for AA. The analysis of statistical data substantiates that this HPTLC method can be used for the simultaneous estimation of AN and AA in biological samples. Copyright © 2015 John Wiley & Sons, Ltd.
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TLC can provide both a phytochemical fingerprint and when compared to a known standard of artemisinin, can also validate artemisinin content in extracts.
“TLC can provide both a phytochemical fingerprint and when compared to a known standard of artemisinin, can also validate artemisinin content in extracts (Widmer et al. 2007; Khan et al. 2015).”
From Enhancing artemisinin content in and delivery from Artemisia annua: a review of alternative, classical, and transgenic approaches (2021) · cites this paper as Khan et al. 2015
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- Authors
- Khan, S., Ali, A., Ahmad, S., & Abdin, M. Z.
- Affiliations
- Jamia Hamdard (IN)
- Corresponding author
- Abdin, Malik Zainul
- Venue
- Biomedical Chromatography
- Publisher
- Wiley
- Published
- 2015-03-31
- Volume
- 29
- Issue
- 10
- Pages
- 1594-1603
- ISSN
- 0269-3879 (ISSN-L)
- 1099-0801
- OpenAlex
- W2141759090
- Cited by
- 21
- References
- 55
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 4 · 2021: 1 · 2020: 3 · 2018: 4 · 2017: 2 · 2016: 2
- FWCI
- 2.07
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biomedical Chromatography (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Department of Science and Technology, Ministry of Science and Technology (100/IFD/9742/2010-2011)
- Topics
- Physical Sciences › Chemistry › Analytical Chemistry › Chromatography in Natural Products
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Sesquiterpenes and Asteraceae Studies
- Keywords
- artemisinin, artemisinic acid, Artemisia annua, HPTLC method development, simultaneous determination, leaf extract, antimalarial compounds, method validation, detection limit, limit of quantification, UV absorbance detection
- MeSH terms
- Chromatography, High Pressure Liquid, Chromatography, Thin Layer, Sensitivity and Specificity, Ultraviolet Rays, Reproducibility of Results, Artemisia annua, Artemisinins, Limit of Detection
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