摘要
目的通过建立HPLC分析法测定血必净注射液中氨基酸,为提高中药注射液质量标准提供依据。方法通过OPA-FMOC柱前衍生化法,建立同时检测血必净注射液中18种常见氨基酸的液相色谱法。色谱柱为AgilentSB-C18柱(250mm×4.6mm,5μm);流动相A为0.02mol/L醋酸钠溶液(含200μL/L三乙胺和3.5mL/L四氢呋喃,2%醋酸溶液调pH7.2±0.05),流动相B为0.02mol/L醋酸钠溶液-甲醇-乙腈(200:400:400)(pH7.2±0.05),梯度洗脱;体积流量:0～38min为1.0mL/min;柱温40℃;检测波长0～32.5min为338nm,32.5min以后为262nm。结果所测18种氨基酸在各自的高、低质量浓度范围内线性关系良好;3个批次血必净注射液中,游离氨基酸测定平均值为1.320mg/mL,总氨基酸测定平均值为2.027mg/mL。结论所建立的氨基酸定量方法结果准确可靠,重现性好,有助于血必净注射液的质量控制,对中药注射剂安全评价标准的提高具有指导意义。
Objective To enhance the standards of injection of Chinese materia medica （CMM） by establishing a RP-HPLC memod for simultaneous quantitative determination of amino acids in Xuebijing Injection. Methods Liquid chromatographic fingerprint method for simultaneous determination of 18 kinds of common amino acids was established by o-phthalaldenyde-9-fluorenylmethyl chloroformate （OPA-FMOC） pre-column derivatization. The column was Agilent SB-C18 （250mm×4.6mm,5μm）, mobile phase consisted of 0.02 mol/L sodium acetate （A）, including 200 μL/L triethylamine and 3.5 mL/L tetrahydrofuran with pH value （7.2±0.05） regulated by 2% acetic acid solution, and 0.02 mol/L sodium acetate- methanol-acetonitrile （200 ： 400 ： 400） with pH value （7.2 ± 0.05） （B） for gradient elution at the flow rate of 1.0 mL/min within 0--38 rain. The detection wavelength was 338 nm within 0--32.5 min and 262 nm,after 32.5 min, while the column temperature was 40 ℃. Results There wsa a good linear relationship of 18 kinds of amino acids with a range of 0.001 2--0.195 4 mg/mL in both high and low concentration. In three batches of Xuebijing Injections, the mean value was 1.320 mg/mL for free amino acids and 2.027 mg/mL for total amino acids. Conclusion The optimized method is accurate and reproducible, and is suitable for the determination of amino acids in Xuebijing Injection. It contributes to the quality control of the injection and guides the improvement of safety evaluation standard of CMM injections.
出处
《现代药物与临床》
CAS
2011年第1期58-62,共5页
Drugs & Clinic
