白头翁汤中6种指标成分含量测定
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摘要
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(4.6 mm×150 mm,5μm),以乙腈-1 mg/mL磷酸水溶液为流动相进行线性梯度洗脱,检测波长205 nm,流速0.8 mL/min(白头翁皂苷B4)、检测波长335 nm,流速1.0 mL/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=0.999 9)、秦皮乙素在0.92μg～5.52μg(r=0.999 8)、盐酸药根碱在0.7μg～4.2μg(r=0.998 7)、盐酸巴马汀在1.04μg～6.24μg(r=0.997 8)、盐酸小檗碱在2.2μg～13.2μg(r=0.997 9)、白头翁皂苷B4在5.5μg～33μg(r=0.998 0)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 Pulsatilladecoction,high performance liquid chromatography(HPLC)method was established for the determination of 6 components in 4 Chinese herbs(Pulsatillae radix,Coptidis rhizome,Phellodendri cortexand Fraxini cortex)in Pulsatilladecoction.Agilent ZORBAX SB C18 chromatographic columns(4.6 mm×150 mm,5μm)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 1.04-6.24μg(r=0.999 9),esculetin in 0.92-5.52μg(r=0.999 8),jateorhizine hydrochloride in 0.7-4.2μg(r=0.998 7),palmatine chloride in 1.04-6.24μg(r=0.997 8),berberine in 2.2-13.2μg(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 Pulsatillasaponin B4 in Pulsatilladecoction 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 Pulsatilladecoction and related preparations.
In order to formulate more scientific and reasonable quality standard for the clinical use of Pulsatilladecoction,high performance liquid chromatography(HPLC)method was established for the determination of 6 components in 4 Chinese herbs(Pulsatillae radix,Coptidis rhizome,Phellodendri cortexand Fraxini cortex)in Pulsatilladecoction.Agilent ZORBAX SB C18 chromatographic columns(4.6 mm×150 mm,5μm)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 1.04-6.24μg(r=0.999 9),esculetin in 0.92-5.52μg(r=0.999 8),jateorhizine hydrochloride in 0.7-4.2μg(r=0.998 7),palmatine chloride in 1.04-6.24μg(r=0.997 8),berberine in 2.2-13.2μg(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 Pulsatillasaponin B4 in Pulsatilladecoction 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 Pulsatilladecoction and related preparations.
引文
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[2] 赵伟.白头翁汤联合理中汤治疗溃疡性结肠炎的临床疗效[J].中国实用医药,2017,12(9):149-150.
[3] 王忠,陈寅萤,张盈颖,等.多组分多靶点中药药理作用机制研究中的问题和解决策略[J].中国实验方剂学杂志,2018,24(5):1-6.
[4] 郝敏,陆兔林,毛春琴,等.基于中药质量标志物的饮片质量控制研究[J].中草药,2017,48(9):1699-1708.
[5] 胡屹屹,穆祥,胡元亮.白头翁汤及其主要成分对内皮细胞分泌细胞因子TNF-α,TXB2和6-keto-PGF1α的影响[J].吉林农业大学学报,2014(3):342-346.
[6] 梁勇满,赵容,许亮,等.中药白头翁本草考证与中国白头翁属植物分类[J].中国实验方剂学杂志,2017(5):203-209.
[7] 杨洪秀,尹志海.白头翁栽培管理[J].特种经济动植物,2012(4):41-41.
[8] 宋小红,唐建华.“白头翁汤”的作用机制及在兽医临床中的应用[J].现代畜牧科技,2012(1):232-234.
[9] 崔国宁,刘喜平,董俊刚,等.白头翁汤治疗溃疡性结肠炎研究进展[J].山东中医药大学学报,2018,42(4):370-373.
[10] 董兴美.白头翁汤内服与灌肠对溃疡性结肠炎肠黏膜愈合影响[J].中医药临床杂志,2017(9):1491-1493.
[11] 张大鸿,肖生顺.白头翁汤合芍药汤加减治疗细菌性痢疾疗效观察[J].实用中医药杂志,2016,32(12):1160-1161.
[12] 陈振华,管咏梅,杨世林,等.白头翁研究进展[J].中成药,2014,36(11):2380-2383
[13] 刘颖,李世颂,刘娟.HPLC测定关黄柏中生物碱的含量[J].中国实验方剂学杂志,2013,19(16):88-91.
[14] 杨炳友,闫明宇,潘娟,等.秦皮化学成分及药理作用研究进展[J].中医药信息,2016,33(6):116-119.