白头翁汤中6种指标成分含量测定

Determination of Six Constituents in Pulsatilla Decoction

建立高效液相色谱法(HPLC)测定白头翁汤中4味中药(白头翁、黄连、黄柏及秦皮)的6种成分含量的方法。采用安捷伦色谱柱(4.6mm×150mm,5μm),以乙腈-1mg/mL磷酸水溶液为流动相进行线性梯度洗脱,检测波长205nm,流速0.8mL/min(白头翁皂苷B4)、检测波长335nm,流速1.0mL/min(盐酸小檗碱、盐酸药根碱、盐酸巴马汀、秦皮甲素和秦皮乙素)。秦皮甲素在1.04μg^6.24μg(r=0.9999)、秦皮乙素在0.92μg^5.52μg(r=0.9998)、盐酸药根碱在0.7μg^4.2μg(r=0.9987)、盐酸巴马汀在1.04μg^6.24μg(r=0.9978)、盐酸小檗碱在2.2μg^13.2μg(r=0.9979)、白头翁皂苷B4在5.5μg^33μg(r=0.9980)范围内线性关系良好。秦皮甲素、秦皮乙素、盐酸药根碱、盐酸巴马汀、盐酸小檗碱和白头翁皂苷B4的加样回收率分别98.74%、98.12%、97.90%、96.95%、97.43%和98.45%,RSD均在2.0%以下,秦皮甲素、秦皮乙素、盐酸药根碱、盐酸巴马汀、小檗碱和白头翁皂苷B4在白头翁汤供试品中的含量分别为0.7%、0.36%、0.12%、0.38%、8.26%和4.71%。所建立的方法重复性、线性及稳定性等良好,准确灵敏,可用于白头翁汤及相关制剂的质量控制。

In order to formulate more scientific and reasonable quality standard for the clinical use of Pulsatilla decoction,high performance liquid chromatography (HPLC) method was established for the determination of 6 components in 4 Chinese herbs ( Pulsatillae radix,Coptidis rhizome,Phellodendri cortex and Fraxini cortex ) in Pulsatilla decoction.Agilent ZORBAX SB C18 chromatographic columns ( 4.6 mm× 150 mm,5 μm) were used;The linear gradient elution was performed with acetonitrile-1 mg/mL phosphoric acid solution;The detection wavelength was 205 nm,the flow rate was 0.8 mL/min (anemoside B4),the detection wavelength was 335 nm,and the flow rate was 1.0 mL/min (aesculin,esculetin,jateorhizine hydrochloride,palmatine chloride,berberine).The results showed that aesculin was in the range of 1.04 -6.24 μg (r=0.999 9), esculetin in 0.92-5.52 μg (r=0.999 8), jateorhizine hydrochloride in 0.7- 4.2 μg (r=0.998 7), palmatine chloride in 1.04-6.24 μg (r=0.997 8),berberine in 2.2-13.2 μg (r= 0.997 9 ), and anemoside B4 in 5.5-33μg (r=0.998 0).The recoveries of aesculin, esculetin,jateorhizine hydrochloride, palmatine chloride,berberine,and anemoside B4 were 98.74%,98.12%,97.90%,96.95%,97.43%,and 98.45%,respectively.All RSD were less than 2.0%.The contents of aesculin,aesculetin,jateorhizine hydrochloride,palmatine chloride,berberine and Pulsatilla saponin B4 in Pulsatilla decoction were 0.7%,0.36%,0.12%,0.38%,8.26% and 4.71% respectively.The established method has good repeatability,linearity and stability,is accurate and sensitive,and can be used for the quality control of Pulsatilla decoction and related preparations.