Quantitative Analysis of Triterpenoids Content in Centella asiatica (L.) Urban. by Ultra Performance Liquid Chromatography (UPLC)

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Jiranuch Mingmuang
Vilailak Chuennang-chee
Apirak Sakpetch
Somchit Niumsakul
Sakwichai Onthong

Abstract

Gotu Kola (Centella asiatica (L.) Urb., Apiaceae) is an herb on the National List of Essential Medicine. The major substances in Gotu Kola is a group of triterpenes and derivatives with various biological activities. The objective of this study is to develop analytical methods for quantification of asiaticoside, madecassoside, asiatic acid and madecassic acid in Gotu Kola raw material and extract using the Ultra Performance Liquid Chromatographic technique by extracting the powdered drug or dissolve the Gotu Kola extract with methanol. A 4 ml – portion of sample solution was injected to UPLC system by using AcquityTM UPLC column BEH C18, 2.1 x 50 mm, 1.7 mm and using distilled water and acetonitrile as mobile phase, flow rate 0.6 ml/min and measured with a PDA at a wavelength of 205 nm. Linearity was established for asiaticoside, asiatic acid, and madecassic acid concentration range of 0.01-0.1 mg/ml and 0.03-0.21 mg/ml for madecassoside with a coefficient of determination (R2) 0.999. The % recovery was in the range of 95-105%. HORRAT was in the range of 0.06-0.3. LOD and LOQ of asiaticoside, madecassoside, asiatic acid and madecassic acid were determined, the LOD were equals 0.0021, 0.0080, 0.0114 and 0.01309 mg/ml, respectively and LOQ were equals 0.0081, 0.0283, 0.0410 and 0.0473 mg/ml, respectively. It was shown that this developed method was suitable and could be used to analyze the content of triterpenoids content in Gotu Kola. The appropriate specification of triterpenoids content in Gotu Kola could be established.

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References

1. Smitinand T. Thai plant names. Bangkok: Funny publishing company limited; 1980. p. 118. (in Thai)
2. Boonyaprapat N, Chokchaicharoenporn O. Medicinal Plants indigenous to Thailand. Bangkok: Prachachon; 2000. p. 508-15. (in Thai)
3. Hashim P, Sidek H, Helan MHM, Sabery A, Palanisamy UD, Ilhan M. Triterpene composition and bioactivities of Centella asiatica. Molecules. 2011;16:1310-22.
4. Yu QL, Duan HQ, Takaishi Y, Gao WY. A novel triterpene from Centella asiatica. Molecules 2006;11:661-5.
5. Grimaldi R, Ponti FD, Angelo L. D, Caravaggi M, Guidi G, Lecchini S, Frigo G.M, Crema A. Pharmacokinetics of the total triterpenic fraction of Centella asiatica after single and multiple administrations to healthy volunteers. a new assay for asiatic acid. J Ethnopharmacol. 1990;28(2):235-41.
6. National List of Essential Medicines (No. 2) B.E. 2554 (2011). Published in Government Gazette on 28 June B.E. 2554 (2011). (in Thai)
7. Sondari D, Harmami SB, Ghozali M, Randy A, Amanda A.S, Irawan Y. Determination of the active asiaticoside content in Centella asiatica as anti-cellulite agent. Indonesian Journal of Cancer Chemoprevention 2011;2(2):221–6
8. Inamdar P.K., Yeole R.D., Ghogare A.B., Souza de N.J. Determination of biologically active constituents in Centella asiatica. Journal of Chromatography A. 1996;742(1-2):127-30.
9. Tiwari R.K, Chanda S, Deepak M, Murli B, Agarwal A. HPLC method validation for simultaneous estimation of madecassoside, asiaticoside and asiatic acid in Centella asiatica. J Chem Pharm Res. 2010;2(3):223-9.
10. Thongnopnua P. High-performance liquid chromatographic determination of asiatic acid in human plasma. Thai Journal of Pharmaceutical Sciences. 2008;32:10-6.
11. Rafamantanana M.H, Rozet E, Raoelison G.E, Cheuk K, Ratsimamanga S.U, Hubert Ph, Quetin-Leclercq J. An improved HPLC-UV method for the simultaneous quantification of triterpenic glycosides and aglycones in leaves of Centella asiatica (L.) Urb (Apiaceae). J Chromatogr B Analyt Technol Biomed Life Sci. 2009;877(23):2396-402.
12. Jain PK, Agrawal RK. High performance liquid chromatographic analysis of asiaticoside in Centella asiatica (L.) Urban. Chiang Mai Journal of Science. 2008;35(3):521-5.
13. Chamnan Patarapanich, Bungon Kongthong, and Suwanna Laungchonlatan. A HPLC method for determination of asiaticoside, madecassoside, asiatic acid and madecassic acid in the extract of Centella asiatica Linn. development, validation and application. Thai J. Pharm. Sci. 2004;28:42.
14. Laungchonlathan S, Kongthong B, Patarapanich C. A TLC method determination of active constituents, madecassoside and asiaticoside from Centella asiatica Linn. in various seasons in Thailand. Thai J. Pharm. Sci. 2004;28:43.
15. James JT, Dubery IA. Identification and quantification of triterpenoid centelloids in Centella asiatica (L.) Urban by densitometric TLC. Journal of Planar Chromatography-Modern TLC. 2011;24(1):82-7.
16. The United States Pharmacopoeial Convention. The United States Pharmacopoeia, The National Formulary 39th revision. Rockville, MD.: The United States Pharmacopoeial Convention, Inc.; 2016. p. 6541–7.
17. Department of Medical Sciences. Thai Herbal Pharmacopoeia 2019. Bangkok: Prachachon Co., Ltd.; 2019.
18. Department of Medical Sciences. A practical guide for single laboratory method validation of chemical methods. Nonthaburi: Department of Medical Sciences; 2006. (in Thai).