Abstract
Centella asiatica (L.) Urban is a traditional herbal medicine used in Asiatic countries, and is commonly used to treat various wounds, leprosy, tuberculosis and lupus diseases. In this work, a new method based on microwave assisted extraction followed by liquid chromatography–diode array detection–electrospray ionization multistage tandem mass spectrometry analysis has been developed for the identification and quantification of biologically active constituents in C. asiatica, including asiatic acid, asiaticoside and madecassoside. The separation was performed on an Agilent Eclipse C18 column (150 mm × 4.6 mm i.d., 5 μm) with gradient elution of acetonitrile and 0.1% aqueous acetic acid within 50 min. Detection was performed at 205 nm. The calibration curves showed good linearity (r 2 > 0.9992). The limits of detection ranged from 1.2 to 1.6 μg mL−1. The intra- and inter-day precision was less than 3% and the recovery of the assay was in the range of 95.4–106.8%. The method was successfully applied to the quantification of the three constituents in different samples of C. asiatica. The results indicated that the developed method could be considered to be a simple, rapid and reliable method for the quality evaluation of C. asiatica. The samples were also analyzed on a liquid chromatography–electrospray ionization–time-of-flight mass spectrometry system to confirm the identification results.
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References
Randriamampionona D, Diallo B, Rakotoniriana F, Rabemanantsoa C, Cheuk K, Corbisier AM, Mahillon J, Ratsimamanga S, Jaziri ME (2007) Fitoterapia 78:482–489. doi:10.1016/j.fitote.2007.03.016
Pan J, Kai GQ, Yuan CX, Jin RS (2007) Chromatographia 66:121–123. doi:10.1365/s10337-007-0243-5
Jia GT, Lu XY (2008) J Chromatogr A 1193:136–141. doi:10.1016/j.chroma.2008.04.024
Sampson JH, Raman A, Karlsen G, Navsaria H, Leigh IM (2001) Phytomedicine 8:230–235. doi:10.1078/0944-7113-00032
Macquart FX, Chastang F, Simeon A, Birembaut P, Gillery P, Wegrowski Y (1999) Eur J Dermatol 9:289–296
Shukla A, Rasik AM, Jain GK, Shankar R, Kulshrestha DK, Dhawan BN (1999) J Ethnopharmacol 65:1–11. doi:10.1016/S0378-8741(98)00141-X
Shukla A, Rasik AM, Dhawan BN (1999) Phytother Res 13:50–54. doi:10.1002/(SICI)1099-1573(199902)13:1<50::AID-PTR368>3.0.CO;2-V
Cheng CL, Koo MWL (2000) Life Sci 67:2647–2653. doi:10.1016/S0024-3205(00)00848-1
kumar MHV, Gupta YK (2002) J Ethnopharmacol 79:253–260
Zhang T, Lee TZ, Qi SH, Xu YB, Xie JL (2005) Chin J Exp Surg 22:43–45
Babu TD, Kuttan G, Padikkala J (1995) J Ethnopharmacol 48:53–57. doi:10.1016/0378-8741(95)01284-K
Chen SW, Wang WJ, Li WJ, Wang R, Li YL, Huang YN, Liang X (2006) Pharmacol Biochem Behav 85:339–344. doi:10.1016/j.pbb.2006.08.017
Gnanapragasam A, Ebenezar KK, Sathish V, Govindaraju P, Devaki T (2004) Life Sci 76:585–597. doi:10.1016/j.lfs.2004.09.009
Gnanapragasam A, Yogeeta S, Subhashini R, Ebenezar KK (2007) Mol Cell Biochem 294:55–63. doi:10.1007/s11010-006-9245-0
Inamdar EK, Yeole RD, Ghogare AB, de Souza NJ (1996) J Chromatogr A 742:127–130. doi:10.1016/0021-9673(96)00237-3
Shen Y, Han C, Liu JD, Liu AL, Liu CP (2008) Chromatographia 68:679–682. doi:10.1365/s10337-008-0731-2
Chemat F, Lucchesi ME, Smadja J, Faretto L, Colnadhi G, Visinoni F (2006) Anal Chim Acta 555:157–160. doi:10.1016/j.aca.2005.08.071
Morales S, Canosa P, Rubí E, Cela R (2005) J Chromatogr A 1082:128–135. doi:10.1016/j.chroma.2005.05.059
Lu Y, Yue XF, Zhang ZQ, Li XX, Wang K (2007) Chromatographia 66:443–446. doi:10.1365/s10337-007-0335-2
Zhang H, Chen ZL, Yang GS, Wang WZ (2008) Food Chem 108:322–328. doi:10.1016/j.foodchem.2007.10.041
Valerie C (2000) Trends Analyt Chem 19:229–248. doi:10.1016/S0165-9936(99)00185-5
Deng CH, Yao N, Wang B, Zhang XM (2006) J Chromatogr A 1103:15–21. doi:10.1016/j.chroma.2005.11.023
Pier LB, Loretta R, Jawahar LS (2002) J Chromatogr A 975:47–70. doi:10.1016/S0021-9673(02)01335-3
Chen JH, Wang FM, Liu J, Lee FSC, Wang XR (2008) Anal Chim Acta 613:184–195. doi:10.1016/j.aca.2008.02.060
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Financial support from the Natural Science Foundation of Zhejiang Province (Y2080331), and Wenzhou Municipal Science and Technology Bureau (H20080052) is gratefully acknowledged.
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Shen, Y., Liu, A., Ye, M. et al. Analysis of Biologically Active Constituents in Centella asiatica by Microwave-Assisted Extraction Combined with LC–MS. Chroma 70, 431–438 (2009). https://doi.org/10.1365/s10337-009-1152-6
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DOI: https://doi.org/10.1365/s10337-009-1152-6