HPLC-PDA法同时测定金嗓子喉片中没食子酸和绿原酸的含量
|
摘要
目的:建立了一种同时测定金嗓子喉片中没食子酸和绿原酸含量的二极管阵列检测器色谱(HPLC-PDA)方法。方法:采用资生堂C18色谱柱(4. 6 mm×250 mm,5μm),以乙腈-0. 1%磷酸水为流动相进行梯度洗脱,流速为1. 0 mL·min~(-1),柱温为25℃,检测波长设为270 nm(没食子酸)和330 nm(绿原酸)。结果:没食子酸与绿原酸浓度分别在10. 02～80. 16μg·mL~(-1)(r=0. 999 8)和5. 973～47. 78μg·mL~(-1)(r=0. 999 7)范围内与峰面积呈良好的线性关系,平均回收率分别为100. 3%(RSD=0. 59%)和99. 6%(RSD=0. 85%),待测液在室温的条件下,60 h之内保持稳定。结论:本方法专属性强,重复性好,结果准确,可用于金嗓子喉片中没食子酸、绿原酸的含量测定。
Objective: High performance liquid chromatography-photodiode array( HPLC-PDA) method was developed for the determination of gallic acid and chlorogenic acid in Golden Throat Lozenge. Methods: The separation of gallic acid and chlorogenic acid was performed on a SHISEIDO HPLC PACKED C_(18) column( 4. 6 mm × 250 mm,5 μm) with the mobile phase consisting of acetonitrile-0. 1% phosphoric acid in a gradient elution mode at a flow rate of 1. 0 mL·min~(-1). Ultraviolet absorption detector was set at 270 nm for gallic acid and 330 nm for chlorogenic acid. The column temperature was set at 25 ℃. Results: Good linearity was obtained in the ranges of 10. 02-80. 16 μg·mL~(-1)( r = 0. 999 8) for gallic acid and 5. 973-47. 78μg·m L~(-1)( r = 0. 999 7) for chlorogenic acid,respectively.The spiked recoveries were 100. 3% and 99. 6% with the relative standard derivations( RSDs) of 0. 59% and 0. 85% for gallic acid and chlorogenic acid,respectively. The limits of quantification( LOQs)( S/N = 10) were 0. 1 μg·mL~(-1) and 0. 06 μg·mL~(-1),and the limits of detection( LODs)( S/N = 3) were 0. 06 μg · mL~(-1) and 0. 01 μg · mL~(-1) for gallic acid and chlorogenic acid,respectively.Conclusion: The results demonstrated that the proposal method is simple,accurate,reproducible and suitable for quality control of Golden Throat Lozenge.
Objective: High performance liquid chromatography-photodiode array( HPLC-PDA) method was developed for the determination of gallic acid and chlorogenic acid in Golden Throat Lozenge. Methods: The separation of gallic acid and chlorogenic acid was performed on a SHISEIDO HPLC PACKED C_(18) column( 4. 6 mm × 250 mm,5 μm) with the mobile phase consisting of acetonitrile-0. 1% phosphoric acid in a gradient elution mode at a flow rate of 1. 0 mL·min~(-1). Ultraviolet absorption detector was set at 270 nm for gallic acid and 330 nm for chlorogenic acid. The column temperature was set at 25 ℃. Results: Good linearity was obtained in the ranges of 10. 02-80. 16 μg·mL~(-1)( r = 0. 999 8) for gallic acid and 5. 973-47. 78μg·m L~(-1)( r = 0. 999 7) for chlorogenic acid,respectively.The spiked recoveries were 100. 3% and 99. 6% with the relative standard derivations( RSDs) of 0. 59% and 0. 85% for gallic acid and chlorogenic acid,respectively. The limits of quantification( LOQs)( S/N = 10) were 0. 1 μg·mL~(-1) and 0. 06 μg·mL~(-1),and the limits of detection( LODs)( S/N = 3) were 0. 06 μg · mL~(-1) and 0. 01 μg · mL~(-1) for gallic acid and chlorogenic acid,respectively.Conclusion: The results demonstrated that the proposal method is simple,accurate,reproducible and suitable for quality control of Golden Throat Lozenge.
引文
[1] Inoue M,Sakaguchi N,Isuzugawa K,et al. Role of reactive oxygen species in gallic acid-induced apoptosis[J]. Biol Pharm Bull,2000,23(10):1153-1157.
[2]吴艳林,蔡政,张明智,等.没食子酸通过活化线粒体通路诱导CNE-2鼻咽癌移植瘤细胞的凋亡[J].临床与病理杂志,2017,37(6):1124-1129.
[3]张庭秀,马李杰,范贤明,等.没食子酸诱导非小细胞肺癌A549细胞凋亡的机制研究[J].中国现代医学杂志,2018,22(19):16-22.
[4]钟振国,梁红,钟意宁,等.余甘子叶提取成分没食子酸的体外抗肿瘤实验研究[J].时珍国医国药,2009,20(8):1954-1955.
[5] Wang R,Ma L,Weng D,et al. Gallic acid induces apoptosis and enhances the anticancer effects of cisplatin in human small cell lung cancer H446 cell line via the ROS-dependent mitochondrial apoptotic pathway[J]. Oncol Rep,2016,35(5):3075-3083.
[6]刘国卿,王秋娟,杨宏健,等.野葡萄成分没食子酸的药理研究[J].南京药学院学报,1983,22(2):43-47.
[7]谢晓艳,刘洪涛,张吉,等.没食子酸体外抗氧化作用研究[J].重庆医科大学学报,2011,36(3):319-322.
[8]张雅丽,李建科,刘柳.没食子酸的体外抑菌作用研究[J].食品工业科技,2013,34(11):81-84.
[9]李秀丽,廖万清.真菌生物膜研究进展[J].中国真菌学杂志,2006,1(5):318-320.
[10]张浩楠,杨玉梅,邬国栋,等.蒙药广枣抗心律失常作用有效成分的分离[J].包头医学院学报,2013,29(3):1-4.
[11]高雅,李骅,王四旺,等.没食子酸的药理作用及其代谢动力学研究进展[J].西北药学杂志,2014,29(4):435-438.
[12]周志娥,罗秋水,熊建华,等.绿原酸、异绿原酸A对大肠杆菌的抑菌机制[J].食品科技,2014,39(3):228-231.
[13] Li Y,Shi W,Zhou Y,et al. Neuroprotective effects of chlorogenic acid against apoptosis of PC12 cells induced by methylmercury[J]. Environ Toxicol Pharmacol,2008,26(1):13-21.
[14] Jiang Y,Kusama K,Satoh K,et al. Induction of cytotoxicity by chlorogenic acid in human oral tumor cell lines[J]. Phytomedicine,2000,7(6):483-491.
[15]庞美蓉,刘零怡,高汪磊,等.绿原酸调节糖脂代谢的作用机制研究进展[J].中草药,2015,46(2):305-312.
[16]金玲,杜鸿灵,张旭. HPLC波长切换法同时测定正本清茶中没食子酸、绿原酸和栀子苷的含量[J].亚太传统医药,2018,14(1):27-31.
[17]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,2015:197,221.
[18]张莲秀. HPLC法同时测定速效止泻胶囊中没食子酸和绿原酸含量[J].海峡药学,2012,24(10):98-100.
[2]吴艳林,蔡政,张明智,等.没食子酸通过活化线粒体通路诱导CNE-2鼻咽癌移植瘤细胞的凋亡[J].临床与病理杂志,2017,37(6):1124-1129.
[3]张庭秀,马李杰,范贤明,等.没食子酸诱导非小细胞肺癌A549细胞凋亡的机制研究[J].中国现代医学杂志,2018,22(19):16-22.
[4]钟振国,梁红,钟意宁,等.余甘子叶提取成分没食子酸的体外抗肿瘤实验研究[J].时珍国医国药,2009,20(8):1954-1955.
[5] Wang R,Ma L,Weng D,et al. Gallic acid induces apoptosis and enhances the anticancer effects of cisplatin in human small cell lung cancer H446 cell line via the ROS-dependent mitochondrial apoptotic pathway[J]. Oncol Rep,2016,35(5):3075-3083.
[6]刘国卿,王秋娟,杨宏健,等.野葡萄成分没食子酸的药理研究[J].南京药学院学报,1983,22(2):43-47.
[7]谢晓艳,刘洪涛,张吉,等.没食子酸体外抗氧化作用研究[J].重庆医科大学学报,2011,36(3):319-322.
[8]张雅丽,李建科,刘柳.没食子酸的体外抑菌作用研究[J].食品工业科技,2013,34(11):81-84.
[9]李秀丽,廖万清.真菌生物膜研究进展[J].中国真菌学杂志,2006,1(5):318-320.
[10]张浩楠,杨玉梅,邬国栋,等.蒙药广枣抗心律失常作用有效成分的分离[J].包头医学院学报,2013,29(3):1-4.
[11]高雅,李骅,王四旺,等.没食子酸的药理作用及其代谢动力学研究进展[J].西北药学杂志,2014,29(4):435-438.
[12]周志娥,罗秋水,熊建华,等.绿原酸、异绿原酸A对大肠杆菌的抑菌机制[J].食品科技,2014,39(3):228-231.
[13] Li Y,Shi W,Zhou Y,et al. Neuroprotective effects of chlorogenic acid against apoptosis of PC12 cells induced by methylmercury[J]. Environ Toxicol Pharmacol,2008,26(1):13-21.
[14] Jiang Y,Kusama K,Satoh K,et al. Induction of cytotoxicity by chlorogenic acid in human oral tumor cell lines[J]. Phytomedicine,2000,7(6):483-491.
[15]庞美蓉,刘零怡,高汪磊,等.绿原酸调节糖脂代谢的作用机制研究进展[J].中草药,2015,46(2):305-312.
[16]金玲,杜鸿灵,张旭. HPLC波长切换法同时测定正本清茶中没食子酸、绿原酸和栀子苷的含量[J].亚太传统医药,2018,14(1):27-31.
[17]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,2015:197,221.
[18]张莲秀. HPLC法同时测定速效止泻胶囊中没食子酸和绿原酸含量[J].海峡药学,2012,24(10):98-100.