[关键词]
[摘要]
目的 建立滋肾养阴颗粒(ZYG)的定性与定量方法。方法 山茱萸的薄层色谱(TLC)鉴别;以氯仿-甲醇(3:1)为展开剂,5%香草醛-硫酸为显色剂;墨旱莲、泽泻、牡丹皮的TLC鉴别:以石油醚(60~90℃)-三氯甲烷-醋酸乙酯(3:1:2)作为展开剂,采用不同的检视方法,对ZYG中多味药材同时鉴别。采用Boston C18色谱柱(250 mm×4.6 mm,5 μm),以乙腈-0.1%三氟乙酸水溶液为流动相,梯度洗脱,检测波长254 nm,对ZYG中莫诺苷、马钱苷、金丝桃苷、特女贞苷、丹皮酚的量进行测定。结果 可用莫诺苷和马钱苷对ZYG中的山茱萸进行定性鉴别;在同一展开条件下,分别在254 nm下对ZYG中的牡丹皮进行鉴别;在366 nm下对ZYG中的墨旱莲进行鉴别;用5%磷钼酸乙醇为显色剂,于日光下对ZYG中的泽泻进行鉴别;所建立的2个TLC方法,各斑点清晰,Rf值适中,分离度良好。以莫诺苷、马钱苷、金丝桃苷、特女贞苷和丹皮酚为指标成分进行方法学研究,其对应的线性范围分别为4.432~110.8、4.192~104.8、4.040~101.0、4.132~103.3、4.076~101.9 μg/mL,各指标成分的相关系数均大于0.999 7,平均加样回收率在96.57%~98.67%,方法的日内精密度小于2%,日间精密度小于3%。方法的稳定性与重复性良好。结论 该方法准确灵敏,易于操作,专属性强,重复性好,可用于ZYG的质量控制。
[Key word]
[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 (3:1) 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 (3:1:2) 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.
[中图分类号]
[基金项目]