液相層析法對綠云膏貼劑中七種成分之同時分析研究

Simultaneous Analysis of Seven Components in Patch Preparation of Rui-Yun-Gau by High Performance Liquid Chromatography

使用逆相HPLC 模式進行綠云膏貼劑之成分分析，開發出包含黃連中berberine 、coptisine ；黃柏中berberine ；大黃中emodin 、sennoside A ；玄參中harpagoside ；黃芩中baicalin 、baicalein 等七種指標成分之多成分同時定量分析方法。同時以水性基劑及油性基劑等不同製程，探討對綠云膏成分之影響。綠云膏之試料通過保持在30 °C 恒溫之HPLC 層析管（Inertsil ODS-2, 4.6 mm i.d. × 250mm ），移動相採用10% 及60% acetonitrile 之混合溶液（各以磷酸調整pH 值為2.8 ），進行梯度沖提法，以1.0 mL/ 分之流速沖提。使用UV 偵測器，偵測波長設定240 nm 。本法之回收率，同日間及異日間的變異係數均在5% 以下，回收率在90.65~112.22% 之間。這個分析法對於綠云膏製劑中七種指標成分是安定且值得信賴之定量法。A facile HPLC method for the resolution and quantitative measurement of seven marker substances, the active ingredients in patch preparation of Rui-Yun-Gau, was established under the gradient elution in the reversed-phase mode. These marker substances included berberine (Coptidis Rhizoma and Phellodendri Cortex), and coptisine (Coptidis Rhizoma), emodin, and sennoside A (Rhei Rhizoma), harpagoside (Scrophulariae Radix), baicalin, and baicalein (Scutellariae Radix). The ingredients in the water-based and oil-based patches of the formula from different manufactures were also analyzed for quality evaluation. Extracted samples were analyzed with a reversed-phase column (Inertsil 5 ODS-2, 4.6 i.d. × 250 mm) at 30 °C, eluted with a mixture of 10% acetonitrile and 60% acetonitrile aqueous solution (each adjusted to pH 2.8 with phosphoric acid) in gradient manner at a flow-rate of 1.0 mL/min, and detected at 240 nm. Relative coefficients of variations of intra- and inter-day analysis were less than 5%. All the recoveries were 90.65~112.22%, therefore this method could be applied for the simultaneous determination of seven marker substances in “Rui-Yun-Gau”.