Studies on quality specification of Zishen Yangyin Granules / 中草药

ABSTRACT

Objective: To establish methods of qualitative identification and quantitative determination for Zishen Yangyin Granules (ZYG). Methods: The TLC method was used to identify the herb by mixture of chloroform-methanol (31) as a developing solvent on high performance silica gel precoated plate (HSGF254) and using 5% vanillic aldehyde sulfuric acid as a chromogenic reagent for qualitative identification of Corni Fructus; TLC identification of Eclipta prostrata, Alismatis Rhizoma, and Moutan Cortex was performed on high performance silica gel precoated plate (HSGF254) with petroleum ether (60-90 ℃)-chloroform-ethyl acetate (312) as developing solvent. The same developing method was used to identify E. prostrata, A. Rhizoma, and paneol in M. Cortex of ZYG by different detected method at the same time. The contents of morroniside, loganin, hyperoside, specnuezhenide, and paeonol were analyzed by high performance liquid chromatography on C18 column (250 mm × 4.6 mm, 5 μm) with the mobile phase of acetonitrile-0.1% trifluoroacetic acid by gradient elution. The detection wavelength was set at 254 nm. Results: Morroniside and loganin were used to identify C. Fructus in ZYG, paeonol in M. Cortex can be identified at 254 nm; Some substances in E. prostrata can be identified at 366 nm; Some substances in A. Rhizoma can be detected in sunlight, with 5% phosphomolybdic acid in ethanol as a chromogenic reagent. The TLC separation was desirable with moderate Rf value and clear spot. The methodology validation for the assay of morroniside, loganin, hyperoside, specnuezhenide, and paeonol presented that they were in good linear correlation in the ranges of 4.432-110.8, 4.192-104.8, 4.040-101.0, 4.132-103.3, and 4.076-101.9 μg/mL, The correlation coefficients of indicator were over 0.999 7. The average recoveries were between 96.57% and 98.67%. The RSD value of intra-day precision was less than 2% and the RSD value of inter-day precision was less than 3%. The method has good stability and reproducibility. Conclusion: The methods of quality control are specific, reproducible, accurate, and suitable, which can be successfully applied to the quality control of ZYG.