铁皮石斛专属性成分的分离制备及质量标准研究

Separation and identification of specific components and quality standard of stem of Dendrobium officinale

采用色谱和光谱技术首次从铁皮石斛追踪分离并鉴定专属性成分佛莱心苷。对铁皮石斛药材粉末特征进行描述。采用薄层色谱法对铁皮石斛中佛莱心苷进行定性分析,硅胶SGF254薄层板,展开剂乙酸乙酯-丁酮-甲酸-水(4∶3∶1∶1),显色剂三氯化铝,结果铁皮石斛中佛莱心苷的薄层色谱鉴别特征明显,具有一定专属性,可实现铁皮石斛药材与易混淆的齿瓣石斛、杯鞘石斛、兜唇石斛等石斛的加工品区分鉴别。对铁皮石斛的粉末特征进行描述。采用高效液相色谱法对铁皮石斛药材中的活性成分柚皮素进行定量分析,Bischoff Chromatography HIPAK NC-04 ODS AB色谱柱(4.4 mm×250 mm,5μm),流动相乙腈-0.1%磷酸梯度洗脱,检测波长226 nm,柱温25℃;结果柚皮素的定量测定在3.90~250.00 mg·L^(-1)呈良好的线性关系,r=0.999 9;平均回收率99.20%(RSD 0.17%),20批铁皮石斛药材质量分数在0.190~0.498 mg·g^(-1)。所建立的定性和定量方法简单灵敏,准确可靠,重现性好,为提高铁皮石斛的质量控制方法提供了实验依据,对于保证其质量和临床疗效具有重要意义。

The violanthin,a specific component,was separated and identified from the stems of Dendrobium officinale by chromatographic technique and spectroscopic method for the first time. The microscopic characteristics of D. officinale powder were examined under a microscopy and described. Thin layer chromatography（ TLC） method was used for qualitative analysis of the violanthin from D. officinale stems with a mixture of ethyl acetate,butanone,formic acid and water（ 4∶ 3∶ 1∶ 1） as the developing solvent on high performance silica gel precoated plate（ SGF254） and using aluminium trichloride as a chromagenic agent. The results showed significant characteristics of violanthin from D. officinale stems on TLC,with certain specificity,and could be used to distinguish it from other easily confusing processed medicinal stems of D. devonianum,D. gratiosissimum and D. aphyllum. The content of naringenin,an active ingredient in D. officinale stems was determined by HPLC analysis on a Bischoff Chromatography HIPAK NC-04 ODS AB column（ 4. 4 mm × 250 mm,5 mm） with acetonitrile-0. 1% phosphoric acid solution as the mobile phase for gradient elution. The wavelength was set at 226 nm and column temperature was 25 ℃. The HPLC method showed good linearity within the range of 3. 90-250. 00 g·m L^-1（ r = 0. 999 9） for naringenin. The average recovery of naringenin was 99. 20% with 0. 17% of RSD. The mass fraction of 20 batches of D. officinale stems was between 0. 190 and 0. 498 mg·g- 1. The established qualitative and quantitative method was simple and rapid with good repeatability and accuracy,providing experimental basis for improving the quality standard of D. officinale,with a very important significance to ensure its quality and clinical effect.