HPLC法同时测定苏叶黄连汤中5种成分的含量分析
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摘要
目的:建立同时测定苏叶黄连汤中木犀草素、芹菜素、迷迭香酸、盐酸小檗碱和盐酸巴马汀5种成分的HPLC方法。方法:采用Agilent Hypersil BDS-C_(18)色谱柱(4.6 mm×250 mm, 5μm);流动相为乙腈(C)-0.2%磷酸水溶液(三乙胺调节pH=4)(A),梯度洗脱;流速1.0 mL·min~(-1);检测波长345 nm;柱温35℃。结果:5种成分分离度良好,阴性无干扰,木犀草素、芹菜素、迷迭香酸、盐酸小檗碱和盐酸巴马汀的质量浓度分别在0.13～4.20μg·mL~(-1)(r~2=0.999 8)、0.05～1.50μg·mL~(-1)(r~2=0.999 7)、8.75～280.00μg·mL~(-1)(r~2=0.999 5)、28.75～920.00μg·mL~(-1)(r~2=0.999 7)和1.63～52.00μg·mL~(-1)(r~2=0.999 5)范围内与峰面积呈良好的线性关系;精密度、重复性良好,RSD均小于3%;在室温条件下24 h内稳定;平均加样回收率在99.41%～102.81%之间(RSD为0.27%～2.88%)。结论:所建立的方法操作简单,重复性好,可用于苏叶黄连汤中5种成分的同时测定。
Objective: To develop an HPLC method for the simultaneous content determination of luteolin, apigenin, rosmarinic acid, berberine hydrochloride and palmatine hydrochloride in suye huanglian decoction. Method: The chromatography conditions were defined as Agilent Hypersil BDS-C_(18) column(4.6 mm×250 mm,5 μm), with the mobile phase comprising of acetonitrile(C)-0.2% phosphatic acid(pH=4)(A) flowing at 1.0 mL·min~(-1) in a gradient elution manner; the detection wavelengths were set at 345 nm, The temperature was 35 ℃. Results: Five components were well seperated with good linearity in their own ranges without disturbing peaks from negative samples, luteolin is 0.13～4.20μg·mL~(-1)(r~2=0.999 8), apigenin is 0.05～1.50 μg·mL~(-1)(r~2=0.999 7), rosmarinic acid is 8.75～280.00 μg·mL~(-1)(r~2=0.999 5), berberine hydrochloride is 28.75～920.00 μg·mL~(-1)(r~2=0.999 7) and palmatine hydrochloride is 1.63～52.00 μg·mL~(-1)(r~2=0.999 5), respectively. The accuracy and repeatability were satisfactory with RSD values less than 3%, the stability was good in 24 h. The average recoveries fell between 99.41%～102.81%(RSD with 0.27%～2.88%). Conclusion: The method is simple, rapid and reproducible, and can be used for the simultaneous content determination.
Objective: To develop an HPLC method for the simultaneous content determination of luteolin, apigenin, rosmarinic acid, berberine hydrochloride and palmatine hydrochloride in suye huanglian decoction. Method: The chromatography conditions were defined as Agilent Hypersil BDS-C_(18) column(4.6 mm×250 mm,5 μm), with the mobile phase comprising of acetonitrile(C)-0.2% phosphatic acid(pH=4)(A) flowing at 1.0 mL·min~(-1) in a gradient elution manner; the detection wavelengths were set at 345 nm, The temperature was 35 ℃. Results: Five components were well seperated with good linearity in their own ranges without disturbing peaks from negative samples, luteolin is 0.13～4.20μg·mL~(-1)(r~2=0.999 8), apigenin is 0.05～1.50 μg·mL~(-1)(r~2=0.999 7), rosmarinic acid is 8.75～280.00 μg·mL~(-1)(r~2=0.999 5), berberine hydrochloride is 28.75～920.00 μg·mL~(-1)(r~2=0.999 7) and palmatine hydrochloride is 1.63～52.00 μg·mL~(-1)(r~2=0.999 5), respectively. The accuracy and repeatability were satisfactory with RSD values less than 3%, the stability was good in 24 h. The average recoveries fell between 99.41%～102.81%(RSD with 0.27%～2.88%). Conclusion: The method is simple, rapid and reproducible, and can be used for the simultaneous content determination.
引文
[1] 李京津, 宋平. 从脾胃升降理论探讨苏叶黄连汤加味治疗慢性肾功能衰竭氮质血症新思路[J]. 西部中医药, 2016, 29(1):54-57.
[2] 刘岩, 吕美. 加味苏叶黄连汤治疗妊娠恶阻26例[J]. 湖南中医杂志, 2012, 28(3):56.
[3] 姬李岩, 张建伟. 浅析苏叶黄连汤在妊娠恶阻中的应用[J]. 江西中医药, 2017, 48(10):78-80.
[4] 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,2015:318.
[5] 王维琪. 黄连有效成分的药理研究与进展[J]. 中医临床研究, 2016, 8(26):147-148.
[6] 王利红, 唐文照, 辛义周. 黄连中生物碱成分及药理作用研究进展[J]. 山东中医药大学学报, 2015(4):389-392.
[7] Zhang Y, Li X, Zhang Q, et al. Berberine hydrochloride prevents postsurgery intestinal adhesion and inflammation in rats[J]. Journal of Pharmacology & Experimental Therapeutics, 2014, 349(3):417-426.
[8] Ji C, Zhang DF, Li AH, et al. Effect of berberine hydrochloride on grass carp Ctenopharyngodon idella, serum bactericidal activity against Edwardsiella ictaluri[J]. Fish Shellfish Immunol, 2012, 33(1):143-145.
[9] 杨慧, 马培, 林明宝,等. 紫苏叶化学成分、抗炎作用及其作用机制研究进展[J]. 中国药理学与毒理学杂志, 2017, 31(3):279-286.
[10] Yu H, Feng QJ, Juan ML, et al. Phytochemical and phytopharmacological review of Perilla frutescens L. (Labiatae), a traditional edible-medicinal herb in China[M]. Food & Chemical Toxicology An International Journal Published for the British Industrial Biological Research Association, 2016.
[11] 刘浏. 紫苏叶的研究进展[J]. 中国医学创新, 2012, 9(6):162-164.
[12] Kim DH, Kim YC, Choi UK. Optimization of antibacterial activity of Perilla frutescens var. acuta leaf against Staphylococcus aureus using evolutionary operation factorial design technique[J]. International Journal of Molecular Sciences, 2011, 12(4):2395-2407.
[13] Zhu F, Asada T, Sato A, et al. Rosmarinic acid extract for antioxidant, antiallergic, and α-glucosidase inhibitory activities, isolated by supramolecular technique and solvent extraction from Perilla leaves[J]. Journal of Agricultural & Food Chemistry, 2014, 62(4):885-892.
[14] 魏科, 李玲, 宁毅,等. 中药材对流感病毒神经氨酸酶活性影响的统计分析[C]. 医药学会免疫学分会第二次学术交流会暨中华中医药学会免疫学分会青年委员会成立大会, 2016.
[15] 孙响波, 于妮娜, 张法荣. 苏叶黄连汤治疗慢性肾衰竭作用机制探讨[J]. 现代中西医结合杂志, 2014, 23(8):895-896.
[16] 喻樊. HPLC法同时测定濒危植物——中华补血草中异槲皮苷、桑色素、槲皮素、木犀草素和芹菜素的含量[J]. 药物分析杂志, 2014, 857(4):632-635.
[17] 侯晓军, 唐菱, 袁敏,等. HPLC法同时测定止吐软膏中小檗碱和巴马汀的含量[J]. 应用化工, 2016, 45(4):781-783.
[18] 张超. 关黄柏不同炮制品中盐酸小檗碱与盐酸巴马汀含量测定[J]. 山东中医药大学学报, 2013(1):73-76.
[19] 李翔, 刘皈阳, 马建丽,等. HPLC法同时测定扁咽口服液中盐酸巴马汀和盐酸小檗碱的含量[J]. 解放军药学学报, 2013(5):458-460.
[20] 丘振文, 钟瑜. HPLC法测定腹达康微丸中盐酸小檗碱的含量[J]. 中药新药与临床药理, 2013, 24(6):616-618.
[21] 安杉杉, 陈燕忠, 林世源,等. HPLC测定脱脂紫苏子中迷迭香酸和木犀草素的含量[J]. 中国实验方剂学杂志, 2014, 20(22):66-68.
[22] 李彩虹, 彭罡, 周克元. RP-HPLC定量三乙胺法测定中药黄连中盐酸小檗碱的含量[J]. 东南大学学报(医学版), 2012, 31(1):8-11.
[23] 张玲容. HPLC测定白带丸中盐酸小檗碱的含量[J]. 中华中医药学刊, 2011,29(4):905-906.
[24] 赵曦, 杨东生, 张瑶,等. HPLC法同时测定清热治痢丸中6种成分[J]. 中成药, 2017, 39(5):956-959.
[25] 刘吉成. HPLC测定黑草中木犀草素和芹菜素的含量[J]. 中国实验方剂学杂志, 2012, 18(20):72-74.
[2] 刘岩, 吕美. 加味苏叶黄连汤治疗妊娠恶阻26例[J]. 湖南中医杂志, 2012, 28(3):56.
[3] 姬李岩, 张建伟. 浅析苏叶黄连汤在妊娠恶阻中的应用[J]. 江西中医药, 2017, 48(10):78-80.
[4] 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,2015:318.
[5] 王维琪. 黄连有效成分的药理研究与进展[J]. 中医临床研究, 2016, 8(26):147-148.
[6] 王利红, 唐文照, 辛义周. 黄连中生物碱成分及药理作用研究进展[J]. 山东中医药大学学报, 2015(4):389-392.
[7] Zhang Y, Li X, Zhang Q, et al. Berberine hydrochloride prevents postsurgery intestinal adhesion and inflammation in rats[J]. Journal of Pharmacology & Experimental Therapeutics, 2014, 349(3):417-426.
[8] Ji C, Zhang DF, Li AH, et al. Effect of berberine hydrochloride on grass carp Ctenopharyngodon idella, serum bactericidal activity against Edwardsiella ictaluri[J]. Fish Shellfish Immunol, 2012, 33(1):143-145.
[9] 杨慧, 马培, 林明宝,等. 紫苏叶化学成分、抗炎作用及其作用机制研究进展[J]. 中国药理学与毒理学杂志, 2017, 31(3):279-286.
[10] Yu H, Feng QJ, Juan ML, et al. Phytochemical and phytopharmacological review of Perilla frutescens L. (Labiatae), a traditional edible-medicinal herb in China[M]. Food & Chemical Toxicology An International Journal Published for the British Industrial Biological Research Association, 2016.
[11] 刘浏. 紫苏叶的研究进展[J]. 中国医学创新, 2012, 9(6):162-164.
[12] Kim DH, Kim YC, Choi UK. Optimization of antibacterial activity of Perilla frutescens var. acuta leaf against Staphylococcus aureus using evolutionary operation factorial design technique[J]. International Journal of Molecular Sciences, 2011, 12(4):2395-2407.
[13] Zhu F, Asada T, Sato A, et al. Rosmarinic acid extract for antioxidant, antiallergic, and α-glucosidase inhibitory activities, isolated by supramolecular technique and solvent extraction from Perilla leaves[J]. Journal of Agricultural & Food Chemistry, 2014, 62(4):885-892.
[14] 魏科, 李玲, 宁毅,等. 中药材对流感病毒神经氨酸酶活性影响的统计分析[C]. 医药学会免疫学分会第二次学术交流会暨中华中医药学会免疫学分会青年委员会成立大会, 2016.
[15] 孙响波, 于妮娜, 张法荣. 苏叶黄连汤治疗慢性肾衰竭作用机制探讨[J]. 现代中西医结合杂志, 2014, 23(8):895-896.
[16] 喻樊. HPLC法同时测定濒危植物——中华补血草中异槲皮苷、桑色素、槲皮素、木犀草素和芹菜素的含量[J]. 药物分析杂志, 2014, 857(4):632-635.
[17] 侯晓军, 唐菱, 袁敏,等. HPLC法同时测定止吐软膏中小檗碱和巴马汀的含量[J]. 应用化工, 2016, 45(4):781-783.
[18] 张超. 关黄柏不同炮制品中盐酸小檗碱与盐酸巴马汀含量测定[J]. 山东中医药大学学报, 2013(1):73-76.
[19] 李翔, 刘皈阳, 马建丽,等. HPLC法同时测定扁咽口服液中盐酸巴马汀和盐酸小檗碱的含量[J]. 解放军药学学报, 2013(5):458-460.
[20] 丘振文, 钟瑜. HPLC法测定腹达康微丸中盐酸小檗碱的含量[J]. 中药新药与临床药理, 2013, 24(6):616-618.
[21] 安杉杉, 陈燕忠, 林世源,等. HPLC测定脱脂紫苏子中迷迭香酸和木犀草素的含量[J]. 中国实验方剂学杂志, 2014, 20(22):66-68.
[22] 李彩虹, 彭罡, 周克元. RP-HPLC定量三乙胺法测定中药黄连中盐酸小檗碱的含量[J]. 东南大学学报(医学版), 2012, 31(1):8-11.
[23] 张玲容. HPLC测定白带丸中盐酸小檗碱的含量[J]. 中华中医药学刊, 2011,29(4):905-906.
[24] 赵曦, 杨东生, 张瑶,等. HPLC法同时测定清热治痢丸中6种成分[J]. 中成药, 2017, 39(5):956-959.
[25] 刘吉成. HPLC测定黑草中木犀草素和芹菜素的含量[J]. 中国实验方剂学杂志, 2012, 18(20):72-74.