RESUMO
Objective To investigate the spectrum-activity relationship between HPLC fingerprint of Ilex latifolia from 10 batches in different regions and its inhibition of xanthine oxidase (XOD) efficacy and to reveal the material basis of I. latifolia. Methods The samples from 10 batch different regions were determined with the HPLC method. The similarity of the representative standard fingerprint was evaluated using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (Version 2004A). The common pattern was established for the principal component analysis (PCA). The HPLC method was used to establish the enzyme-substrate reaction system, and the inhibitory activity of XOD of 10 batches of I. latifolia produced in different areas was studied. The SPSS analysis correlation was used to conduct correlation analysis based on spectrum-activity relationship. The model of mouse hyperuricemia were induced by potassium oxonate, and the levels of UC and XOD in plasma and liver were detected. Results Fifteen common peaks in HPLC-fingerprint of I. latifolia were obtained. It was tentatively concluded that peak 11 (ilekudinoside E) and peak 14 (kudinoside F) had the moderate correlation with the inhibitory effect of XOD in the fifteen characteristic peaks. Peaks 5, 6 (kaempferol-3-O-β-D-rutinoside), 7 (isorhamnetin-3-O-β-D-rutinoside), 10 (latifoloside C), and 13(latifoloside D) had the strongest correlation. The quality of I. latifolia was influenced by the concentration of latifoloside H (peak 9) obtained by PCA. In the vivo experiment of hyperuricemia model mice, the levels of plasma UC and XOD of the mice in different doses of I. latifolia extracts groups were reduced, and the activities of XOD in livers tissue were also reduced. Conclusion Ten batches of samples from different habitats had potent inhibitory effects on XOD. There may be a certain relationship between seven compounds showed in HPLC fingerprint and its inhibitory effect on XOD.
RESUMO
Objective: Based on the study of chemical composition research of Ilex latifolia, to develop an HPLC method for the simultaneous determination of latifoloside F, latifoloside H, latifoloside C, latifoloside D, kudinoside F and ilekudinoside E. The purity of the six saponins was confirmed by LC-MS. Methods: Chromatographic separation was performed on a Wonda Cract ODS-2 column (250 mm × 4.6 mm, 5 μm) with linear gradient elution of acetonitrile and water at a flow rate of 1 mL/min, and the detection wavelength was 210 nm. The mass spectrum was negative ion mode, the capillary temperature was 250℃, the solvent gas was nitrogen, the flow rate was 0.75 L/min, the spray voltage was 4.0 kV, and the lens voltage was 125V. Results: The six saponins were well separated and all calibration curves showed good linearity within the test ranges (r > 0.999 2). The average recoveries were between 99.52% and 100.96%, and RSD values were less than 2%. Conclusion: An accurate and simple method is first established for the qualitation and quantification of six saponins in I. latifolia. The results demonstrate that the method could be applied as a reliable quality control method for the seed of I. latifolia.