Box-Behnken experimental design for extraction of artemisinin from Artemisia annua and validation of the assay method

Abstract

Obtaining artemisinin and its derivatives is very costly, which limits access to low-income people. Some hydroalcoholic extract of Artemisia annua L., Asteraceae, which has shown comparable antimalarial activity could be an alternative to the purified compound, especially if the cultivars have higher content of artemisinin. The objective of this study was to evaluate the effects of the extraction parameters (ethanol graduation, previous shaking time in an ultrasound bath and drug/solvent ratio) on the yield of artemisinin in the liquid extract obtained by percolation from A. annua and then optimize the extraction efficiency of this compound. The Box-Behnken (33) factorial design was used in association with response surface methodology. The derivatization reaction of artemisinin was used in an analytical method which was submitted to validation, after reaching the specification of the selectivity, linearity, precision, accuracy and robustness. Detection and quantification limits were 1.3 and 4.0 μg/ml, respectively. The largest amount of this compound of interest was obtained without any ultrasound bath, with an ethanol graduation of 95% and a drug/solvent ratio of 2%. Drug/solvent ratio was the factor which most influenced extraction efficiency. The maximum range of artemisinin yield was 1.21%. Information obtained in this study can be used for future approaches to determining and extracting artemisinin from A. annua.

References

  1. Atemnkeng, M.A., Chimanuka, B., Dejaegher, B., Heyden, Y.V., Plaizier-Vercammen, J., 2009. Evaluation of Artemisia annua L. infusion efficacy for the treatment of malaria in Plasmodium chabaudi chabaudi infected mice. Exp. Parasitol. 122, 344–348.

    Article  PubMed  Google Scholar 

  2. Bilia, A.R., Magalhaes, P.M., Bergonzi, M.C., Vincieri, F.F., 2006. Simultaneous analysis of artemisinin and flavonoids of several extracts of Artemisia annua L. obtained from a commercial sample and a selected cultivar. Phytomedicine 13, 487–493.

    CAS  Article  PubMed  Google Scholar 

  3. Briars, R., Paniwnyk, L., 2013. Effect of ultrasound on the extraction of artemisinin from Artemisia annua L. Ind. Crop Prod. 42, 595–600.

    CAS  Article  Google Scholar 

  4. Brisibe, E.A., Chukwurah, P.N., 2014. Production of artemisinin in plant and in microbial systems need not be mutually exclusive. In: Aftab, T., Ferreira, J.F.S., Khan, M.M.A., Naeem, M. (org.), Artemisia annua - Pharmacology and Biotechnology, Berlin: Springer-Verlag Heidelberg, pp. 269–292.

    Google Scholar 

  5. Celeghini, R.M.S., Sousa, I.M.O., Silva, A.P., Rodrigues, R.A.F., Foglio, M.A., 2009. Development and validation of analytical methodology by HPLC-IR for evaluation of artemisinin on Artemisia annua L. Quim. Nova 32, 875–878.

    CAS  Article  Google Scholar 

  6. Delabays, N., Simonnet, X., Gaudin, M., 2001. The genetics of artemisinin content in Artemisia annua L. and the breeding of high yielding cultivars. Curr. Med. Chem. 8, 1795–1801.

    CAS  Article  PubMed  Google Scholar 

  7. Diawara, H.Z., Gbaguidi, F., Gbenou, J., Laleye, A., Semde, R., Some, I., Sinsin, B., Moudachirou, M., Leclercq, J.Q., Evrard, B., 2012. Conseil africain et malgache pour l’enseignement superieur - CAMES. Formulation of oral pharmaceutical dosage forms containing crude extracts of Artemisia annua L. Pharmacopée et medicine traditionnelle africaine, v.16., http://publication.lecames.org/index.php/pharm/article/view/27 (accessed February 2014).

  8. Diawara, H.Z., Gbaguidi, F., Evrard, B., Leclercq, J.Q., Moudachirou, M., Debrus, B., Hubert, P., Rozet, E., 2011. Validation, transfer and measurement uncertainty estimation of an HPLC-UV method for the quantification of artemisinin in hydro alcoholic extracts of Artemisia annua L. J. Pharm. Biomed. Anal. 56, 7–15.

    CAS  Article  PubMed  Google Scholar 

  9. Erbay, Z., Icier, F., 2009. Optimization of hot air drying of olive leaves using response surface methodology. J. Food Eng. 91, 533–541.

    CAS  Article  Google Scholar 

  10. Erdemoglu, N., Orhan, I., Kartal, M., Adiguzel, N., Bani, B., 2007. Determination of artemisinin in selected Artemisia L. species of Turkey by reversed phase HPLC. Rec. Nat. Prod. 1, 36–43.

    CAS  Google Scholar 

  11. Fleming, A., Freyhold, M., 2007. Assessing the Technical and Economic Viability of the Ethanolic Extraction of Artemisia annua L. with Special Reference to Tanzania, http://www.mmv.org/sites/default/files/uploads/docs/publications/6%20-%203 _ethanolic-extraction-december-2007.pdf (accessed February 2014).

    Google Scholar 

  12. Hao, J., Han, W., Huang, S., Xue, B., Deng, X., 2002. Microwave-assisted extraction of artemisinin from Artemisia annua L. Sep. Purif. Technol. 28, 191–196.

    CAS  Article  Google Scholar 

  13. ICH, 2009. Harmonised Tripartite Guideline. Guidance for Industry: Q8(R2), Pharmaceutical Development, International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). U.S. Department of Health and Human Services, Food and Drug Administration, Rockville.

    Google Scholar 

  14. Klayman, D.L., 1985. Qinghaosu (artemisinin): an antimalarial drug from China. Science 228, 1049–1055.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  15. Elford, B.C., Roberts, M.F., Phillipson, D., Wilson, R.J.M., 1987. Potentiation of the antimalarial activity of qinghaosu by methoxylated flavones. Trans. R. Soc. Trop. Med. Hyg. 81, 434–436.

    CAS  Article  PubMed  Google Scholar 

  16. Liu, C., Zhou, H., Zhao, Y., 2007. An effective method for fast determination of artemisinin in Artemisia annua L. by high performance liquid chromatography with evaporative light scattering detection. Anal. Chim. Acta 581, 298–302.

    CAS  Article  PubMed  Google Scholar 

  17. Marchese, J.A., Rehder, V.L.G., Sartoratto, A., 2001. Quantificação de artemisinina em Artemisia annua L. - uma comparação entre as técnicas de cromatografia em camada delgada com detecção densitométrica e cromatografia de alta eficiência com detecção no ultravioleta. Rev. Bras. Plantas Med. 4, 81–87.

    CAS  Google Scholar 

  18. Meshnick, S.R., Taylor, T.E., Kamchonwongpaisan, S., 1996. Artemisinin and the antimalarial endoperoxides: from herbal remedy to targeted chemotherapy. Microbiol. Rev. 60, 301–315.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. Naeem, M., Idress, M., Singh, M., Masroor, M., Moinuddin, A.K., 2014. Artemisia annua: a miraculous herb to cure malaria. In: Aftab, T., Ferreira, J.F.S., Khan, M.M.A., Naeem, M (org.), Artemisia annua - Pharmacology and Biotechnology, Berlin: Springer-Verlag Heidelberg, pp. 27–49.

    Google Scholar 

  20. Ogwang, P.E., Ogwal, J.O., Kasasa, S., Ejobi, F., Kabasa, D., Obua, C., 2011. Use of Artemisia annua L. infusion for malaria prevention: mode of action and benefits in a Ugandan community. Br. J. Pharm. Res. 1, 124–132.

    CAS  Article  Google Scholar 

  21. Rehder, V.L.G., Rodrigues, M.V.N., Sartoratto, A., Foglio, M.A., 2002. Dosagem de artemisinina em Artemisia annua L. por cromatografia líquida de alta eficiência com detecção por índice de refração. Rev. Bras. Farmacogn. 12, 116–118.

    Article  Google Scholar 

  22. Rodrigues, R.A.F., Foglio, M.A., Júnior, S.B., Santos, A.S., Rehder, V.L.G., 2006. Optimization of the extraction and isolation of the antimalarial drug artemisinin from Artemisia annua L. Quim. Nova 29, 368–372.

    CAS  Article  Google Scholar 

  23. Portal Action, 2014. Experimento de Box-Behnken, http://www.portalaction.com.br/649-632-experimento-de-box-behnken (accessed August 2014).

    Google Scholar 

  24. Suberu, J., Song, L., Slade, S., Sullivan, N., Barder, G., Lapkin, A.A., 2013. A rapid method forthe determination of artemisinin and its biosynthetic precursors in Artemisia annua L. crude extracts. J. Pharm. Biomed. Anal. 84, 269–277.

    CAS  Article  PubMed  Google Scholar 

  25. US-FDA, 1994. United States Food and Drug Administration. Reviewer Guidance - Validation of Chromatographic Methods. Center of Drug Evaluation and Research, Silver Spring.

    Google Scholar 

  26. Van der Kooy, F., 2014. Reverse pharmacology and drug discovery: Artemisia annua and its anti-HIV activity. In: Aftab, T., Ferreira, J.F.S., Khan, M.M.A., Naeem, M. (org.), Artemisia annua - Pharmacology and Biotechnology. Berlin: Springer-Verlag Heidelberg, pp. 249–267.

    Google Scholar 

  27. Yang, B., Liu, X., Gao, Y.X., 2009. Extraction optimization of bioactive compounds (crocin, geniposide and total phenolic compounds) from Gardenia (Gardenia jas-minoides Ellis) fruits with response surface methodology. Innov. Food Sci. Emerg. Technol. 10, 610–615.

    CAS  Article  Google Scholar 

  28. WHO, 2012. Effectiveness of Non-Pharmaceutical Forms of Artemisia annua L. against malaria. Global Malaria Programme. WHO Position Statement, World Health Organization, Switzerland.

    Google Scholar 

  29. WHO, 2006. WHO monograph on good agricultural and collection practices (GACP) for Artemisia annua L. WHO Library, World Health Organization, Switzerland.

    Google Scholar 

  30. Wright, C.W., Linley, P.A., Brun, R., Wittlin, S., Hsu, E., 2010. Ancient Chinese methods are remarkably effective forthe preparation ofartemisinin-rich extracts of Qing Hao with potent antimalarial activity. Molecules 15, 804–812.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  31. Zhao, S.S., Zeng, M.Y., 1986. Determination of qinghaosu in Artemisia annua L. by high performance liquid chromatography. Anal. Chem. 58, 289–292.

    Article  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Maria Teresa F. Bara.

Additional information

Authors contributions

EOS: acquisition of the data, analysis and interpretation of the data, drafting of the article; LLB: statistical expertise, drafting of the article; ECC: conception and design, provision of study materials; MTFB: conception and design, provision of study materials, obtaining of funding, drafting of the article.

Rights and permissions

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

de Silva, E.O., Borges, L.L., da Conceição, E.C. et al. Box-Behnken experimental design for extraction of artemisinin from Artemisia annua and validation of the assay method. Rev. Bras. Farmacogn. 27, 519–524 (2017). https://doi.org/10.1016/j.bjp.2017.03.002

Download citation

Keywords

  • Medicinal plant
  • Artemisinin
  • Extraction process
  • Box-Behnken factorial design
  • Response surface methodology
  • HPLC