Free accessResearch articleFirst published online 2013-12
Development and Validation of a Modified Ultrasound-Assisted Extraction Method and a HPLC Method for the Quantitative Determination of Two Triterpenic Acids in Hedyotis diffusa
In the present study, the oleanolic acid (OA) and ursolic acid (UA) contents of Hedyotis diffusa and H. corymbosa were determined by a rapid, selective and accurate method combining modified ultrasound-assisted extraction (MUAE) and HPLC. Compared with traditional extraction methods, MUAE reduced the extraction time, the extraction temperature and the solvent consumption and maximized the extraction yields of OA and UA. Furthermore, the combined MUAE–HPLC method was applied to quantitate OA and UA in plant samples and exhibited good repeatability, reproducibility and stability. The mean recovery studies (one extraction cycle) for OA and UA were between 91.3 and 91.7% with RSD values less than 4.5%. The pioneer method was further applied to quantitate OA and UA in six samples of H. diffusa and five samples of H. corymbosa. The results showed that the OA and UA content in the samples from different sources were significantly different. This report is valuable for the application of H. diffusa and H. corymbosa obtained from different regions in clinical research and pharmacology.
XuG.H., KimY.H., ChiS.W., ChooS.J., RyooI.J., AhnJ.S., YooI.D. (2010) Evaluation of human neutrophil elastase inhibitory effect of iridoid glycosides from Hedyotis diffusa. Bioorganic & Medicinal Chemistry Letters, 20, 513–515.
2.
WuP.K., TaiW.C.S., LiangZ.T., ZhaoZ.Z., HsiaoW.L.W. (2009) Oleanolic acid isolated from Oldenlandia diffusa exhibits a unique growth inhibitory effect against ras-transformed fibroblasts. Life Sciences, 85, 113–121.
3.
KimY., ParkE.J., KimJ., KimY., KimS.R., KimY.Y. (2001) Neuroprotective constituents from Hedyotis diffusa. Journal of Natural Products, 64, 75–78.
4.
AttardE., AttardH. (2006) The potential angiotensin-converting enzyme inhibitory activity of oleanolic acid in the hydroethanolic extract of Crataegus monogyna Jacq. Natural Product Communications, 1, 381–385.
5.
YanS.L., HuangC.Y., WuS.T., YinM.C. (2010) Oleanolic acid and ursolic acid induce apoptosis in four human liver cancer cell lines. Toxicology in Vitro, 24, 842–848.
6.
ZhangY.X., KongC.Z., WangH.Q., WangL.H., XuC.L., SunY.H. (2009) Phosphorylation of Bcl-2 and activation of caspase-3 via the c-Jun N-terminal kinase pathway in ursolic acid-induced DU145 cells apoptosis. Biochimie, 91, 1173–1179.
7.
RamosA.A., Pereira-WilsonC., CollinsA.R. (2010) Protective effects of ursolic acid and luteolin against oxidative DNA damage include enhancement of DNA repair in Caco-2 cells. Mutation Research, 692, 6–11.
8.
OvesnáZ., KozicsK., SlameňováD. (2006) Protective effects of ursolic acid and oleanolic acid in leukemic cells. Mutation Research, 600, 131–137.
9.
HuangM.T., HoC.T., WangZ.Y., FerraroT., LouY.R., StauberK., MaW., GeorgiadisC., LaskinJ.D., ConneA.H. (1994) Inhibition of skin tumorigenesis by rosemary and its constituents carnosol and ursolic acid. Cancer Research, 54, 701–708.
10.
YimE.K., LeeK.H., NamkoongS.E., UmS.J., ParkJ.S. (2006) Proteomic analysis of ursolic acid-induced apoptosis in cervical carcinoma cells. Cancer Letters, 235, 209–220.
11.
HsuY.L., KuoP.L., LinC.C. (2004) Proliferative inhibition, cell-cycle disregulation, and induction of apoptosis by ursolic acid in human non-small cell lung cancer A549 cells. Life Sciences, 75, 2303–2316.
12.
YangY.C., WeiM.C., HuangT.C. (2012) Optimisation of an ultrasound-assisted extraction followed by RP-HPLC separation for the simultaneous determination of oleanolic acid, ursolic acid and oridonin content in Rabdosia rubescens. Phytochemical Analysis, 23, 627–636.
13.
YangY.C., WeiM.C., HongS.J., HuangT.C., LeeS.Z., (2013) Development/optimization of a green procedure with ultrasound-assisted improved supercritical carbon dioxide to produce extracts enriched in oleanolic acid and ursolic acid from Scutellaria barbata D. DonIndustrial Crops and Products, 49, 542–553.
14.
FangX., WangJ., YuX., ZhangG., ZhaoJ. (2010) Optimization of microwave-assisted extraction followed by RP-HPLC for the simultaneous determination of oleanolic acid and ursolic acid in the fruits of Chaenomeles sinensis. Journal of Separation Science, 33, 1147–1155.
15.
ClaudeB., MorinP., LafosseM., AndreP. (2004) Evaluation of apparent formation constants of pentacyclic triterpene acids complexes with derivatized beta- and gamma-cyclodextrins by reversed phase liquid chromatography. Journal of Chromatography A, 1049, 37–42.
16.
DominguesR.M.A., SousaG.D.A., FreireC.S.R., SilvestreA.J.D., Pascoal NetoC. (2010) Eucalyptus globulus biomass residues from pulping industry as a source of high value triterpenic compounds. Industrial Crops and Products, 31, 65–70.
17.
XiaE.Q., YuY.Y., XuX.R., DengG.F., GuoY.J., LiH.B. (2012) Ultrasound-assisted extraction of oleanolic acid and ursolic acid from Ligustrum lucidum Ait. Ultrasonics Sonochemistry, 19, 772–776.
18.
YangY.C., WeiM.C., LianF.Y., HuangT.C. (2013) Simultaneous extraction and quantitation of oleanolic acid and ursolic acid from Scutellaria barbata D. Don by ultrasound-assisted extraction and high-performance liquid chromatography. Chemical Engineering Communications, 201DOI: 10.1080/ 00986445.2013.777901.
19.
WeiM.C., YangY.C., ChiuH.F., HongS.J. (2013) Development of a hyphenated procedure of heat-reflux and ultrasound-assisted extraction followed by RP-HPLC separation for the determination of three flavonoids content in Scutellaria barbata D. Don, Journal of Chromatography. B Analytical Technologies in the Biomedical and Life Sciences, 940, 126–134.
20.
YangY.C., WeiM.C., HuangT.C., LeeS.Z., LinS.S. (2013) Comparison of modified ultrasound-assisted and traditional extraction methods for the extraction of baicalin and baicalein from Radix Scutellariae. Industrial Crops and Products, 45, 182–190.
21.
ZhouC., ShengY., ZhaoD., WangZ., TaoJ. (2010) Variation of oleanolic and ursolic acid in the flesh of persimmon fruit among different cultivars. Molecules, 15, 6580–6587.
