A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of six sesquiterpene lactones: 3β-[3-(4-hydroxyphenyl)acetyl-β-D-glucopyranosyioxy] -8yβ-hydroxy-4(15),10(14)11(13)-guaiatrien-12,6-olide (AR1) 3β-[3-(4-hydroxyphenyl)acetyl-β-D-glucopyranosyloxy]-4(15),10(14) 11(13)-guaiatrien-12,6-olide (AR2), 3β-(β-D- glucopyranosyloxy)-8β-(4-hydroxyphenyl)acetoxy-4(15),10(14)11(13) -guaiatrien-12,6-olide (AR3), 3β-(β-D-glucopyranosyloxy)- 8β-(2-hydroxy-3-methylbutanoyloxy)acetoxy-4(15),10(14)11(13)-guaiatrien-12, 6-olide (AR4), 3β-(β-D-glucopyranosyloxy)-4(15),10(14) 11(13)-guaiatrien-12,6-olide (AR5), and 3β-hydroxy-4(15),10(14) 11(13)-guaiatrien-12,6-olide (AR6) in the whole plant of Ixeris chinensis. The separation by gradient elution was performed on Hypersil ODS-2 column (250 mm x 4.6 mm, 5 μm) at 30°C with acetonitrile and water as the mobile phase, and monitored by absorbance at 238 nm. The parameters of linearity, precision, accuracy, and specificity of the method were evaluated. The recovery of the method is 95.85-98.19%, and linearity (r > 0.9993) was obtained for all sesquiterpene lactones. A high degree of specificity as well as repeatability and reproducibility (relative standard deviation values less than 2.0%) were also achieved. This assay was successfully applied to the determination of six sesquiterpene lactones in ten samples. The results indicated that the developed assay method was rapid, accurate, reliable and could be readily utilized as a quantitative analysis method for I. chinensis.

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