一测多评法测定复方黄柏液涂剂中8种活性成分
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摘要
目的:建立一测多评法同时测定复方黄柏液涂剂中8种活性成分的含量。方法:采用HPLC法,以连翘酯苷A为内参,分别计算其与绿原酸、隐绿原酸、盐酸黄柏碱、芦丁、异绿原酸C、盐酸小檗碱、连翘苷的相对校正因子(RCF),通过RCF计算复方黄柏液涂剂中上述7种成分的含量(计算值),同时采用外标法测定8种成分的含量(实测值),比较计算值与实测值的差异。色谱柱为Agilene TC-C_(18)(250 mm×4. 6 mm,5μm)柱,流动相为乙腈-0. 5%磷酸溶液(梯度洗脱),流速为0. 8 ml·min~(-1),检测波长为277 nm(盐酸小檗碱、连翘苷、盐酸黄柏碱)和325 nm(绿原酸、隐绿原酸、芦丁、异绿原酸C、连翘酯苷A),柱温为30℃,进样量为10μl。结果:绿原酸、隐绿原酸、盐酸黄柏碱、芦丁、异绿原酸C、盐酸小檗碱、连翘酯苷A、连翘苷的线性范围分别为9. 115～182. 300,3. 823～76. 450,0. 258～5. 161,1. 645～32. 910,1. 244～24. 870,2. 002～40. 050,12. 630～252. 600,4. 598～91. 960μg·ml~(-1),r为0. 999 4～0. 999 8;平均回收率分别为97. 1%,97. 7%,96. 9%,98. 3%,97. 5%,98. 1%,98. 3%,96. 9%,RSD为0. 8%～1. 7%(n=6);绿原酸、隐绿原酸、盐酸黄柏碱、芦丁、异绿原酸C、盐酸小檗碱、连翘苷的RCF分别为0. 651,0. 991,1. 384,1. 185,1. 814,0. 632,1. 577,其计算值和实测值之间无明显差异。结论:该方法简单、有效、结果准确、节约成本,可用于复方黄柏液涂剂中上述8种活性成分的同时测定。
Objective: To establish quantitative analysis of multi-components by single marker( QAMS) for the simultaneous determination of 8 active constituents in Fufang Huangbaiye paint. Methods: Using forsythoside A as the internal standard,an HPLC method was used to calculate the relative correction factor( RCF) of forsythoside A to chlorogenic acid,cryptochlorogenic acid,phellodendrine,rutin,isochlorogenic acid C,berberine hydrochloride and phillyrin. The contents of above-mentioned 7 components in Fufang Huangbaiye Paint were calculated by using RCF( calculation values). An external standard method( ESM) was used to determine the contents of 8 constituents( measured values). The differences between the calculation values and the measured values were compared. The determination was performed on an Agilene TC-C_(18)( 250 mm × 4. 6 mm,5 μm) column with the mobile phase consisting of acetonitrile-0. 5% phosphoric acid with gradient elution at the flow rate of 0. 8 ml·min~(-1). The detection wavelength was set at 277 nm for phillyrin,berberinehydrochloride and phellodendrine,and 325 nm for chlorogenic acid,cryptochlorogenic acid,rutin,isochlorogenic acid C and forsythoside A. The column temperature was set at 30℃ and the injection volume was 10 μl. Results: The linear range was 9. 115-182. 300 μg·ml~(-1) for chlorogenic acid,3. 823-76. 450 μg·ml~(-1) for cryptochlorogenic acid,0. 258-5. 161 μg·ml~(-1) for phellodendrine,1. 645-32. 910 μg·ml~(-1) for rutin,1. 244-24. 870 μg·ml~(-1) for isochlorogenic acid C,2. 002-40. 050 μg·ml~(-1) for berberine hydrochloride,12. 630-252. 600 μg · ml~(-1) for forsythoside A and 4. 598-91. 960 μg · ml~(-1) for phillyrin( r =0. 999 4-0. 999 8). The average recovery was 97. 1%,97. 7%,96. 9%,98. 3%,97. 5%,98. 1%,98. 3% and 96. 9%( RSD =0. 8%-1. 7%,n = 6),respectively. RCFs of forsythoside A to chlorogenic acid,cryptochlorogenic acid,phellodendrine,rutin,isochlorogenic acid C,berberine hydrochloride and phillyrin were 0. 651,0. 991,1. 384,1. 185,1. 814,0. 632 and 1. 577,respectively.There were no significant differences between the calculated values and the measured values. Conclusion: The method is simple,effective,accurate and economical,which can be used for the simultaneous determination of 8 active constituents in Fufang Huangbaiye paint.
Objective: To establish quantitative analysis of multi-components by single marker( QAMS) for the simultaneous determination of 8 active constituents in Fufang Huangbaiye paint. Methods: Using forsythoside A as the internal standard,an HPLC method was used to calculate the relative correction factor( RCF) of forsythoside A to chlorogenic acid,cryptochlorogenic acid,phellodendrine,rutin,isochlorogenic acid C,berberine hydrochloride and phillyrin. The contents of above-mentioned 7 components in Fufang Huangbaiye Paint were calculated by using RCF( calculation values). An external standard method( ESM) was used to determine the contents of 8 constituents( measured values). The differences between the calculation values and the measured values were compared. The determination was performed on an Agilene TC-C_(18)( 250 mm × 4. 6 mm,5 μm) column with the mobile phase consisting of acetonitrile-0. 5% phosphoric acid with gradient elution at the flow rate of 0. 8 ml·min~(-1). The detection wavelength was set at 277 nm for phillyrin,berberinehydrochloride and phellodendrine,and 325 nm for chlorogenic acid,cryptochlorogenic acid,rutin,isochlorogenic acid C and forsythoside A. The column temperature was set at 30℃ and the injection volume was 10 μl. Results: The linear range was 9. 115-182. 300 μg·ml~(-1) for chlorogenic acid,3. 823-76. 450 μg·ml~(-1) for cryptochlorogenic acid,0. 258-5. 161 μg·ml~(-1) for phellodendrine,1. 645-32. 910 μg·ml~(-1) for rutin,1. 244-24. 870 μg·ml~(-1) for isochlorogenic acid C,2. 002-40. 050 μg·ml~(-1) for berberine hydrochloride,12. 630-252. 600 μg · ml~(-1) for forsythoside A and 4. 598-91. 960 μg · ml~(-1) for phillyrin( r =0. 999 4-0. 999 8). The average recovery was 97. 1%,97. 7%,96. 9%,98. 3%,97. 5%,98. 1%,98. 3% and 96. 9%( RSD =0. 8%-1. 7%,n = 6),respectively. RCFs of forsythoside A to chlorogenic acid,cryptochlorogenic acid,phellodendrine,rutin,isochlorogenic acid C,berberine hydrochloride and phillyrin were 0. 651,0. 991,1. 384,1. 185,1. 814,0. 632 and 1. 577,respectively.There were no significant differences between the calculated values and the measured values. Conclusion: The method is simple,effective,accurate and economical,which can be used for the simultaneous determination of 8 active constituents in Fufang Huangbaiye paint.
引文
1姚俊,赵霞.复方黄柏液最新临床应用进展[J].中国新药杂志,2014,23(3):308-312,337
2魏晋宝,杨光义,陈欢,等.连翘苷的提取方法、药理毒理及药动学研究进展[J].中国药师,2015,18(12):2144-2148
3陈阳峰,钟晓红.黄柏的药理作用及其活性成分提取[J].作物研究,2015,29(5):564-568
4宋亚玲,王红梅,倪付勇,等.金银花中酚酸类成分及其抗炎活性研究[J].中草药,2015,46(4):490-495
5中国药典[S].2015年版.一部.1244
6王玉团,许丽丽,徐丽华.RP-HPLC-DAD法在复方黄柏液涂剂质量控制方面的研究[J].中国药品标准,2016,17(3):174-178
7潘军.RP-HPLC法同时测定复方黄柏液中盐酸小檗碱、绿原酸和连翘苷的含量[J].齐鲁药事,2009,28(9):524-526
8王智民,钱忠直,张启伟,等.一测多评法建立的技术指南[J].中国中药杂志,2011,36(6):657-658
9胡丹.HPLC-QAMS同时测定复方黄芩片中6种活性成分的含量[J].中国药房,2018,29(11):1510-1514
10陈芳,邓雁如,郭利平.HPLC法同时测定金银花及金芪降糖片中9种成分的含量[J].天津中医药,2014,31(3):168-172
11陈建琴.UPLC法测定抗感解毒颗粒中欧前胡素、连翘苷、连翘酯苷A的含量[J].中国药师,2015,18(11):1876-1878
2魏晋宝,杨光义,陈欢,等.连翘苷的提取方法、药理毒理及药动学研究进展[J].中国药师,2015,18(12):2144-2148
3陈阳峰,钟晓红.黄柏的药理作用及其活性成分提取[J].作物研究,2015,29(5):564-568
4宋亚玲,王红梅,倪付勇,等.金银花中酚酸类成分及其抗炎活性研究[J].中草药,2015,46(4):490-495
5中国药典[S].2015年版.一部.1244
6王玉团,许丽丽,徐丽华.RP-HPLC-DAD法在复方黄柏液涂剂质量控制方面的研究[J].中国药品标准,2016,17(3):174-178
7潘军.RP-HPLC法同时测定复方黄柏液中盐酸小檗碱、绿原酸和连翘苷的含量[J].齐鲁药事,2009,28(9):524-526
8王智民,钱忠直,张启伟,等.一测多评法建立的技术指南[J].中国中药杂志,2011,36(6):657-658
9胡丹.HPLC-QAMS同时测定复方黄芩片中6种活性成分的含量[J].中国药房,2018,29(11):1510-1514
10陈芳,邓雁如,郭利平.HPLC法同时测定金银花及金芪降糖片中9种成分的含量[J].天津中医药,2014,31(3):168-172
11陈建琴.UPLC法测定抗感解毒颗粒中欧前胡素、连翘苷、连翘酯苷A的含量[J].中国药师,2015,18(11):1876-1878