Abstract
Betulin, oleanolic acid, and betulinic acid are naturally occurring pentacyclic triterpenoids that have significant medicinal value. Considerable amounts of these triterpenoids are available in the outer bark of white birch. In this study, we used ultrasound-assisted extraction (UAE) to extract triterpenoids from birch bark rapidly and with high efficiency. Using high performance liquid chromatography (HPLC), three types of triterpenoids were separated and detected. We examined the differences among triterpenoids extracted from diploid versus tetraploid white birch. Then, we used factor analysis to screen out tetraploid white birches with comprehensively excellent performance. The results indicate that the optimum conditions for extraction include the use of ethanol as an extraction solvent, a solid-to-liquid ratio of 0.1 g/10 ml, ultrasonic power set at 100 W, a temperature of 60 °C and an extraction time of 15 min. A reversed-phase C18 column (4.6 mm × 250 mm × 5 μm) with a column temperature of 30 °C and the mobile phase composed of A (acetonitrile) and B (0.1 % aqueous phosphoric acid, v/v) at a flow rate of 0.5 ml/min were used, and the detection wavelength was 195 nm. No significant difference was observed between diploid and tetraploid white birch in terms of the content of three types of triterpenoids (at a confidence level of 0.05). As triterpenoid content, height, and DBH (diameter at breast height) are strongly interrelated, we used factor analysis to evaluate all individuals, and we screened out six plus trees with excellent comprehensive characters.
Similar content being viewed by others
References
Alakurtti S, Mäkelä T, Koskimies S, Yli-Kauhaluoma J (2006) Pharmacological properties of the ubiquitous natural product betulin. Eur J Pharm Sci 29(1):1–13
Caruso I, Lepore L, De Tommasi N, Dal Piaz F, Frusciante L, Aversano R, Garramone R, Carputo D (2011) Secondary metabolite profile in induced tetraploids of wild Solanum commersonii Dun. Chem Biodivers 8(12):2226–2237
Chen Q, Liu J, Zhang H, He G, Fu M (2009) The betulinic acid production from betulin through biotransformation by fungi. Enzyme Microb Tech 45(3):175–180
Claude B, Morin P, Lafosse M, Andre P (2004) Evaluation of apparent formation constants of pentacyclic triterpene acids complexes with derivatized β- and γ-cyclodextrins by reversed phase liquid chromatography. J Chromatogr A 1049(1–2):37–42
De Jesus-Gonzalez L, Weathers PJ (2003) Tetraploid Artemisia annua hairy roots produce more artemisinin than diploids. Plant Cell Rep 21(8):809–813
Dhawan OP, Lavania UC (1996) Enhancing the productivity of secondary metabolites via induced polyploidy: a review. Euphytica 87(2):81–89
Dolan JW (2002) Peak tailing and resolution. LCGC North Am 20(5):430–436
Fu JY, Qian LB, Zhu LG, Liang HT, Tan YN, Lu HT, Lu JF, Wang HP, Xia Q (2011) Betulinic acid ameliorates endothelium-dependent relaxation in L-NAME-induced hypertensive rats by reducing oxidative stress. Eur J Pharm Sci 44(3):385–391
Fujioka T, Kashiwada Y, Kilkuskie RE, Cosentino LM, Ballas LM, Jiang JB, Janzen WP, Chen IS, Lee KH (1994) Anti-AIDS agents, 11. Betulinic acid and platanic acid as anti-HIV principles from Syzigium claviflorum, and the anti-HIV activity of structurally related triterpenoids. J Nat Prod 57(2):243–247
Guo S, Duan J, Tang Y, Su S, Shang E, Ni S, Qian D (2009) High-performance liquid chromatography—two wavelength detection of triterpenoid acids from the fruits of Ziziphus jujuba containing various cultivars in different regions and classification using chemometric analysis. J Pharmaceut Biomed 49(5):1296–1302
Huang L, Li J, Ye H, Li C, Wang H, Liu B, Zhang Y (2012) Molecular characterization of the pentacyclic triterpenoid biosynthetic pathway in Catharanthus roseus. Planta 236(5):1571–1581
Kim Y, Hahn E, Murthy HN, Paek K (2004) Effect of polyploidy induction on biomass and ginsenoside accumulations in adventitious roots of ginseng. Journal of Plant Biology 47(4):356–360
Krasutsky PA (2006) Birch bark research and development. Nat Prod Rep 23(6):919–942
Kromidas S, Kuss HJ (2009) Quantification in LC and GC: a practical guide to good chromatographic data. Wiley-VCH, Weinheim, p 8
Lavania UC, Srivastava S, Lavania S, Basu S, Misra NK, Mukai Y (2012) Autopolyploidy differentially influences body size in plants, but facilitates enhanced accumulation of secondary metabolites, causing increased cytosine methylation. Plant J 71(4):539–549
Liu J (1995) Pharmacology of oleanolic acid and ursolic acid. J Ethnopharmacol 49(2):57–68
Madlung A (2013) Polyploidy and its effect on evolutionary success: old questions revisited with new tools. Heredity (Edinb) 110(2):99–104
MobileReference (ed) (2008) The illustrated encyclopedia of trees and shrubs: an essential guide to trees and shrubs of the world. Boston, MobileReference
Ng DW, Zhang C, Miller M, Palmer G, Whiteley M, Tholl D, Chen ZJ (2011) cis- and trans-Regulation of miR163 and target genes confers natural variation of secondary metabolites in two Arabidopsis species and their allopolyploids. Plant Cell 23(5):1729–1740
Pai SR, Nimbalkar MS, Pawar NV, Dixit GB (2011) Optimization of extraction techniques and quantification of betulinic acid (BA) by RP-HPLC method from Ancistrocladus heyneanus Wall. Ex Grah. Ind Crop Prod 34(3):1458–1464
Patočka J (2003) Biologically active pentacyclic triterpenes and their current medicine signification. J Appl Biomed 1:7–12
Pisha E, Chai H, Lee IS, Chagwedera TE, Farnsworth NR, Cordell GA, Beecher CW, Fong HH, Kinghorn AD, Brown DM, Et A (1995) Discovery of betulinic acid as a selective inhibitor of human melanoma that functions by induction of apoptosis. Nat Med 1(10):1046–1051
Pollier J, Goossens A (2012) Oleanolic acid. Phytochemistry 77:10–15
Ranney TG (2006) Polyploidy: from evolution to new plant development. Combined proceedings international plant propagators’ society, vol 56, pp 137–142
Salin O, Alakurtti S, Pohjala L, Siiskonen A, Maass V, Maass M, Yli-Kauhaluoma J, Vuorela P (2010) Inhibitory effect of the natural product betulin and its derivatives against the intracellular bacterium Chlamydia pneumoniae. Biochem Pharmacol 80(8):1141–1151
Schinor EC, Salvador MJ, Turatti IC, Zucchi OL, Dias DA (2004) Comparison of classical and ultrasound-assisted extractions of steroids and triterpenoids from three Chresta spp. Ultrason Sonochem 11(6):415–421
Shizhen L (2011) Compendium of materia medica. Jiangsu People’s Publishing House, Nanjing, pp 519–520 (in Chinese)
Tang J, Li J, Qi W, Qiu W, Li P, Li B, Song B (2011) Inhibition of SREBP by a small molecule, Betulin, improves hyperlipidemia and insulin resistance and reduces atherosclerotic plaques. Cell Metab 13(1):44–56
Trivedi PC (2006) Medicinal plants: traditional knowledge. I.K.International Publishing House Pvt, New Delhi, p 240
Wood M (2008) The earthwise herbal: a complete guide to old world medicinal plants. North Atlantic Books, Berkeley, pp 139–142
Xu R, Fazio GC, Matsuda SPT (2004) On the origins of triterpenoid skeletal diversity. Phytochemistry 65(3):261–291
Yogeeswari P, Sriram D (2005) Betulinic acid and its derivatives: a review on their biological properties. Curr Med Chem 12(6):657–666
Yu S (2008) Triterpenoid chemistry. Chemical Industry Press, Beijing, pp 1–3, 119–128, (in Chinese)
Zhang Z (2008) Dendrology. China Forestry Publishing House, Beijing, pp 194–196 (in Chinese)
Acknowledgments
The authors thank ELIXIGEN (SHANGHAI) CO., LTD. for copyediting this manuscript to eliminate grammatical and spelling errors.
Author information
Authors and Affiliations
Corresponding author
Additional information
Project funding: This work was financially supported by National Forestry Department Public Benefit Research Foundation of China (201204302).
The online version is available at http://www.springerlink.com
Corresponding editor: Hu Yanbo
Rights and permissions
About this article
Cite this article
Wang, S., Zhao, H., Jiang, J. et al. Analysis of three types of triterpenoids in tetraploid white birches (Betula platyphylla Suk.) and selection of plus trees. J. For. Res. 26, 623–633 (2015). https://doi.org/10.1007/s11676-015-0096-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11676-015-0096-y