Abstract
A simple procedure has been described for simultaneous determination and improved yield of three pentacyclic triterpenoids—betulinic, oleanolic, and ursolic acids—from callus cultures of Lantana camara. Cell biomass was obtained from leaf disk explants cultured on Murashige and Skoog (Physiol Plant 15:473–497, 1962) medium supplemented with 5 μM 6-benzylaminopurine, 1 μM 2,4-dichlorophenoxyacetic acid, and 1 μM α-naphthaleneacetic acid. Optimum separation of the three compounds was achieved by reverse-phase high-pressure liquid chromatography on a C18 column with 80:20 (v/v) acetonitrile/water as mobile phase. With this route, a yield of 3.1% betulinic acid, 1.88% oleanolic acid, and 4.12% ursolic acid per gram dry weight was obtained from cultures. Leaves from the parent plant, used as control, showed total absence of betulinic acid, and the quantities of oleanolic and ursolic acids present in them were only marginally higher than that found in in vitro-raised cultures. Presence of the three compounds was further confirmed by electrospray ionization mass spectrometry.
Similar content being viewed by others
References
Cichewicz R. H.; Kouzi S. A. Chemistry, biological activity, and chemotherapeutic potential of betulinic acid for the prevention and treatment of cancer and HIV infection. Med Res Rev 24: 90–114; 2004.
Ghisalberti E. L. Lantana camara L. Verbenaceae. Fitoterapia 71: 467–486; 2000.
Liu J. Oleanolic acid and ursolic acid: research perspectives. J Ethnopharmacol 100: 92–94; 2005.
Murashige T.; Skoog F. A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15: 473–497; 1962.
Naik S. K.; Pattnaik S.; Chand P. K. In vitro propagation of pomegranate (Punica granatum L. cv. Ganesh) through axillary shoot proliferation from nodal segments of mature tree. Sci Hortic 79: 175–183; 1999.
Pasqua G.; Silvestrini A.; Monacelli B.; Mulinacci N.; Menendez P.; Botta B. Triterpenoids and ellagic acid derivatives from in vitro cultures of Camptotheca acuminata Decaisne. Plant Physiol Biochem 44: 220–225; 2006.
Razborsek M. I.; Voncina D. B.; Dolecek V.; Voncina E. Determination of oleanolic, betulinic and ursolic acid in Lamiaceae and mass spectral fragmentation of their trimethylsilylated derivatives. Chromatographia 67: 433–440; 2008.
Rout G. R.; Saxena C.; Samantaray S.; Das P. Rapid clonal propagation of Plumbago zeylanica Linn. Plant Growth Regul 28: 1–4; 1999.
Sharma O. P.; Singh A.; Sharma S. Levels of lantadenes, bioactive pentacyclic triterpenoids in young and mature leaves of Lantana camara var. aculeata. Fitoterapia 71: 487–491; 2000.
Skrzypek Z.; Wysokinska H. Sterols and triterpenes in cell culture of Hyssopus officinalis L. Z Naturforsch 58: 308–312; 2003.
Zhao G.; Yan W.; Cao D. Simultaneous determination of betulin and betulinic acid in white birch bark using RP-HPLC. J Pharma Biomed Anal 43: 959–962; 2007.
Author information
Authors and Affiliations
Corresponding author
Additional information
Editor: S. J. Murch
Rights and permissions
About this article
Cite this article
Srivastava, P., Chaturvedi, R. Simultaneous determination and quantification of three pentacyclic triterpenoids—betulinic acid, oleanolic acid, and ursolic acid—in cell cultures of Lantana camara L.. In Vitro Cell.Dev.Biol.-Plant 46, 549–557 (2010). https://doi.org/10.1007/s11627-010-9298-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-010-9298-3