高效毛细管电泳法测定热毒宁注射液中阴阳离子和小分子有机酸成分的含量  

Determination of Zwitterion and Low-Molecular-Weight Acid in Re-Du-Ning Injection by HPCE

作　　者：葛雯 杨彪[2] 于洋[4] 康经武 李海波[2] 王振中[2] Ge Wen;Yang Biao;Yu Yang;Kang Jingwu;Li Haibo;Wang Zhenzhong(College of Traditional Chinese Medicine,Zhejiang Pharmaceutical College,Ningbo 315100,China;State Key Laboratory of New-tech for Chinese Medicine Pharmaceutical Process,Kanion Pharmaceutical Co.,Ltd.,Lianyungang 222001,China;State Key Laboratory of Bioorganic and Natural Products Chemistry,Shanghai institute of Organic Chemistry,Chinese Academy of Sciences,Shanghai 200032,China;Institute of Traditional Chinese Medicine and Natural Products,Jinan University,Guangzhou 510632,China)

机构地区：[1]浙江医药高等专科学校中药学院,宁波315100 [2]江苏康缘药业股份有限公司中药制药过程新技术国家重点实验室,连云港222001 [3]中国科学院上海有机化学研究所生命有机化学国家重点实验室,上海200032 [4]暨南大学中药及天然药物研究所,广州510632

出　　处：《世界科学技术-中医药现代化》2020年第11期3913-3918,共6页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家中医药管理局中药标准化专项(ZYBZH-C-JS-31):热毒宁注射液标准化建设,负责人:萧伟。

摘　　要：目的建立高效毛细管电泳法(High Performance Capillary Electrophoresis,HPCE)同时测定热毒宁注射液中阴阳离子和小分子有机酸成分的含量测定方法。方法以弹性石英毛细管柱(50μm×60cm,有效长度49.5 cm)为分离通道,0.25 mmol·L^(-1)十六烷基三甲基氯化铵(CTAC)-20 mmol·L^(-1)邻苯二甲酸氢钾溶液(pH 5.8)为缓冲溶液,分离电压-12 kV,柱温为25℃,0.6 psi压力进样6 s,于232 nm波长下检测,12种阴离子及小分子有机酸10 min内能够完全分离;6 mmol·L^(-1)α-羟基异丁酸-12 mmol·L^(-1)咪唑(pH 4.0)为缓冲溶液,分离电压15 kV,柱温为25℃,0.6 psi压力进样8 s,于214 nm波长下检测,6种阳离子10 min内能够完全分离。结果热毒宁注射液中阴、阳离子和小分子有机酸在测定的质量浓度范围内均具有良好的线性关系(r≥0.9900)。K^(+)、Na^(+)、Cl^(-)和L^(-)苹果酸的平均回收率分别为89.1%、106.8%、120.1%、120.7%,RSD值分别为1.57%、7.19%、4.1%、2.8%。10批次热毒宁注射液中K+、Na^(+)、Cl^(-)和L-苹果酸的平均含量分别为387.68、3941.25、1595.33、690.58μg·mL^(-1)。结论该方法简单准确,具有良好的重复性和稳定性,可为热毒宁注射液质量标准的完善提供科学依据。Objective To establish a high-performance capillary electrophoresis method for the simultaneous determination of inorganic anions, inorganic cations, and low-molecular-weight acid in Re-Du-Ning injection.Methods The Chromatographic separation was performed on an elastic quartz capillary column(50 μm × 60 cm) with an effective length of 50 cm. Twelve inorganic anions and low-molecular-weight acids in Re-Du-Ning injection were completely separated within 10 minutes with the electrolyte solution of 0.25 mmol·L^(-1) CTAC and 20 mmol·L^(-1) Potassium biphthalate(p H 5.8), the separation voltage of-12 k V, the column temperature of 25℃, the detection wavelength of 232 nm,the injection pressure of 0.6 psi, and the injection time of 6 seconds. Six inorganic cations in Re-Du-Ning injection were completely separated within 10 minutes with the electrolyte solution of 6 mmol·L^(-1)α-hydroxyisobutric acid and12 mmol·L-1 Imidazole(pH 4.0), the separation voltage of 15 kV, the column temperature of 25℃, the detection wavelength of 214 nm, the injection pressure of 0.6 psi and the injection time of 8 seconds.Results All calibration curves of the ions and low-molecular-weight acids showed excellent linear regressions(r ≥ 0.9900) within the test range. The average recoveries and RSD of K^(+), Na^(+), Cl^(-)and L-Malate were 89.1%, 106.8%, 120.1%, 120.7% and1.57%, 7.19%, 4.1%, 2.8%. The average contents of them in 10 batches of Re-Du-Ning injection were 387.68 μg·mL^(-1),3941.25 μg·mL^(-1), 1595.33 μg·mL^(-1), and 690.58 μg·mL^(-1).Conclusion The established method is fast, accurate, and has high repeatability, which could provide references for the quality control of Re-Du-Ning injection.

关 键 词：热毒宁注射液 阴阳离子 小分子有机酸 高效毛细管电泳法 含量测定 

分 类 号：R284.1[医药卫生—中药学]