一测多评法同时测定竭红跌打酊中5种成分的含量
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摘要
目的建立一测多评法同时测定竭红跌打酊中羟基红花黄色素A、儿茶素、表儿茶素、巴西苏木素和(±)原苏木素B的含量。方法采用Agilent Zorbax SB-C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相:乙腈-0.2%磷酸溶液,梯度洗脱;检测波长:403 nm(检测羟基红花黄色素A)和280 nm[检测儿茶素、表儿茶素、巴西苏木素和(±)原苏木素B];流速:0.9 ml/min;柱温:35℃。以儿茶素为内标,建立竭红跌打酊中该成分与羟基红花黄色素A、表儿茶素、巴西苏木素和(±)原苏木素B的相对校正因子,计算其含量。通过外标法测定结果和一测多评法计算结果进行对比,验证所建立方法的可行性。结果羟基红花黄色素A、儿茶素、表儿茶素、巴西苏木素和(±)原苏木素B分别在6.98～139.60、10.12～202.40、5.76～115.2、3.82～76.40、2.18～43.60μg/ml范围内线性关系良好,平均加样回收率分别为98.26%、100.09%、97.79%、97.25%、96.96%,RSD分别为0.88%、0.71%、1.29%、0.85%、1.43%。一测多评法计算结果与外标法测定结果无显著差异。结论一测多评法可以用于竭红跌打酊中5个成分含量的同时测定及质量控制。
Objective To establish a quantitative analysis of multi-components by single-marker(QAMS) method for simultaneous determination of hydroxysafflor ywellow A,catechin,epicatechin,brazilin and(±)protosappanin B in Jiehong Dieda tincture.Methods The analysis was performed on an Agilent Zorbax SB-C_(18) column(4.6 mm×250 mm,5 μm),with the mobile phase comprised of acetonitrile-0.2% phosphoric acid solution flowing at 0.9 ml/min in gradient elution,the column temperature was set at 35 ℃,and the detection wavelengths were set at 403 nm and 280 nm.Catechin was selected as the internal reference substance,and the relative correlation factors of hydroxysafflor yellow A,epicatechin,brazilin and(±)protosappanin B were determined by HPLC.The accuracy and feasibility of the method were validated by comparing the results of QAMS method with those of external standard method.Results The standard curves of hydroxysafflor yellow A,catechin,epicatechin,brazilin and(±)protosappanin B showed a good linearity in 6.98～139.60,10.12～202.40,5.76～115.2,3.82～76.40 and 2.1/8～43.60 μg/ml,respectively.The average recoveries and RSDs were 98.26%(0.88%),100.09%(0.71%),97.79%(1.29%),97.25%(0.85%) and 96.96%(1.43%),respectively.The results obtained by QAMS were similar to those obtained by external standard method.Conclusion The QAMS method can be used for simultaneous the content determination and quality control of 5 components in Jiehong Dieda tincture.
Objective To establish a quantitative analysis of multi-components by single-marker(QAMS) method for simultaneous determination of hydroxysafflor ywellow A,catechin,epicatechin,brazilin and(±)protosappanin B in Jiehong Dieda tincture.Methods The analysis was performed on an Agilent Zorbax SB-C_(18) column(4.6 mm×250 mm,5 μm),with the mobile phase comprised of acetonitrile-0.2% phosphoric acid solution flowing at 0.9 ml/min in gradient elution,the column temperature was set at 35 ℃,and the detection wavelengths were set at 403 nm and 280 nm.Catechin was selected as the internal reference substance,and the relative correlation factors of hydroxysafflor yellow A,epicatechin,brazilin and(±)protosappanin B were determined by HPLC.The accuracy and feasibility of the method were validated by comparing the results of QAMS method with those of external standard method.Results The standard curves of hydroxysafflor yellow A,catechin,epicatechin,brazilin and(±)protosappanin B showed a good linearity in 6.98～139.60,10.12～202.40,5.76～115.2,3.82～76.40 and 2.1/8～43.60 μg/ml,respectively.The average recoveries and RSDs were 98.26%(0.88%),100.09%(0.71%),97.79%(1.29%),97.25%(0.85%) and 96.96%(1.43%),respectively.The results obtained by QAMS were similar to those obtained by external standard method.Conclusion The QAMS method can be used for simultaneous the content determination and quality control of 5 components in Jiehong Dieda tincture.
引文
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[8] 宋粉云,毋福海,钟兆健.HPLC测定万应胶囊中儿茶素、表儿茶素和盐酸小檗碱的含量[J].中成药,2006,28(6):806-810.
[9] 赵焕新,白虹,王元书,等.HPLC测定苏木中的巴西苏木素和原苏木素B[J].华西药学杂志,2010,25(3):363-364.
[10] 陈玉平,毕丹,屠鹏飞.苏木的质量标准研究[J].中国中药杂志,2010,35(16):2068-2071.
[11] 陈雪敏,胡永钢,李美萍,等.高效液相色谱法同时测定苏木中的巴西苏木素和原苏木素 B[J].分析试验室,2012,31(4):98-101.
[2] 国家药典委员会.中华人民共和国药典(一部)[S].北京:中国医药科技出版社,2015:10,151,164.
[3] 周思多,王晓,杨洪军,等.红花药材中四种化学成分的高效液相色谱双波长法同时测定[J].时珍国医国药,2014,25(11):2595-2598.
[4] 姚苗苗,任爱农,董仲才.RP-HPLC法同时测定红花中羟基红花黄色素A与红花黄色素 A的含量[J].药物分析杂志,2010,30(2):263-265.
[5] 张荣,王涛,刘启德,等.高效液相色谱法测定桃仁红花配伍药对中羟基红花黄色素A含量的变化[J].广州中医药大学学报,2009,26(4):381-383.
[6] 张特利,李军山,李振江.HPLC法测定儿茶配方颗粒中儿茶素和表儿茶素的含量[J].中国药房,2015,26(15):2114-2116.
[7] 王淑敏,李惠琳,刘志强,等.儿茶药材中儿茶素和表儿茶素的高效液相色谱法分析研究[J].时珍国医国药,2006,17(4):490-491.
[8] 宋粉云,毋福海,钟兆健.HPLC测定万应胶囊中儿茶素、表儿茶素和盐酸小檗碱的含量[J].中成药,2006,28(6):806-810.
[9] 赵焕新,白虹,王元书,等.HPLC测定苏木中的巴西苏木素和原苏木素B[J].华西药学杂志,2010,25(3):363-364.
[10] 陈玉平,毕丹,屠鹏飞.苏木的质量标准研究[J].中国中药杂志,2010,35(16):2068-2071.
[11] 陈雪敏,胡永钢,李美萍,等.高效液相色谱法同时测定苏木中的巴西苏木素和原苏木素 B[J].分析试验室,2012,31(4):98-101.