Chinese Medicine Clay Poria Quality Control Methods

Rhizoma Heterosmilacis Japonicae is the dried tuber of Heterosmilax japonica Kunth., Heterosmilax yunnanensis Gagnep. or Heterosmilax chinensis Wang.. Rhizoma Heterosmilacis Japonica is used in clinic as an important Traditional Chinese Medicine (TCM). 12 samples of Rhizoma Heterosmilacis Japonica were purchased from TCM shops or straightly collected from the plant habitats in different places of China. The present study provided the methods to isolate constituents, determined the general flavonoids in Rhizoma Heterosmilacis Japonicae by ultraviolet spectrophotometry and determined macrozamin in Rhizoma Heterosmilacis Japonicae by HPLC. HPLC fingerprint chromatogram was also established for Rhizoma Heterosmilacis Japonica. Subsequently, the systematic quality evaluation of Rhizoma Heterosmilacis Japonica was developed.The whole plants of Rhizoma Heterosmilacis Japonicae were extracted with 70% aqueous alcohol to give an alcoholic extract, which was extracted successively with the same volume of petroleum benzene, acetic ether and n-butanol. In order to isolate the constituents in these extracted parts of Rhizoma Heterosmilacis Japonicae, various chromatographic techniques were performed. Six compounds were isolated and identified with UV, IR, NMR and MS data and physical-chemical properties. They wereβ-sitosterol (â… ), daucosterol (â…¡), 3, 3', 5, 5'-tetrahydroxy-4'-methoxystilbene (â…¢), naringenin (â…£),α-L-butylsorbopyranoside (â…¤) and macrozamin (â…¥). The compoundsâ…¢,â…£,â…¤andâ…¥were isolated from this genus for the first time.The concentration of general flavonoids in Rhizoma Heterosmilacis Japonicae was determined with ultraviolet spectrophotometry. Eldrin was standard preparation. The absorbance of general flavonoids in Rhizoma Heterosmilacis Japonicae was determined at 510 nm. The calibration curves of eldrin was linear over the range of 8.80～52.80μg·mL^-1 (r=0.9999, n=6). The mean recovery were 98.3% (RSD=1.2%). The assay which was founded was simple, fast and accurate to measure the concentration of general flavonoids in Rhizoma Heterosmilacis Japonicae. So the assay can be used as one of the method to control the quality of Rhizoma Heterosmilacis Japonicae.The macrozamin in Rhizoma Heterosmilacis Japonicae was determined by reversed phase high performance liquid chromatography (RP-HPLC). The determination was performed on a Century C₁₈ AQ column(250 mm×4.6mm i.d., 5μm) using methanol-water(4: 96, v/v) as the mobile phase at a flow rate of 1.0 ml·min^-1, with a UV detector setting at 215 nm. The calibration curves of macrozamin was linear over the range of 19.12～382.4μg·mL^-1 (r=0.9998, n=6). The mean recovery was 99.5%(RSD=2.1%). The assay was found to be simple and accurate to measure the concentrations of macrozamin in Rhizoma Heterosmilacis Japonicae.The RP-HPLC fingerprint analysis for the quality control of Rhizoma Heterosmilacis Japonicae was established. Kromasil^TM C₁₈ column(250 mm×4.6 mm, 5μm) was used, with mixture of acetonitrile and water as mobile phase in a gradient mode. The concentration of acetonitrile increased from 2% to 3% in 10 minutes, increased to 35% during the later 85 minutes and lasted for 5 minutes. The flow rate was 1.0 mL·min^-1. The wavelength of measurement was 215 nm. 12 batches of Rhizoma Heterosmilacis Japonicae were determined under the condition described above. With the Within-groups linkage method and the Cosine used as the measurement index, the Hierarchical cluster was applied to the fingerprints of Rhizoma Heterosmilacis Japonicae and the 12 batches of Rhizoma Heterosmilacis Japonicae were divided into two grades. Among them the gradeâ… was commendatory. Mutual pattem was established from 5 batches in the gradeâ… and the similarity data were also obtained. These results of the commendatory should be all over 0.90.