反相高效液相色谱法测定射干抗病毒注射液中绿原酸含量

Determination of chlorogenic acid in Shegan Kangbingdu injection by reversed high performance liquid chromatography

目的 建立射干抗病毒注射液中绿原酸的反相高效液相色谱分析方法。方法 色谱柱:Kromasil ODS(5μm,4.6 mm×200 mm),水-甲醇-乙腈-冰醋酸(95∶10∶5∶2,v/v)为流动相,流速1.0 mL·min-1,柱温(23.5±0.1)℃,于326nm测定射干抗病毒注射液中绿原酸含量。结果 线性范围:0.0074～3.7 μg(r=0.9999),绿原酸对照品进样精密度RSD=0.60％,在16 h内峰面积RSD=0.54％,检测限为2.4 ng,定量限为7.4 ng,回收率100.1％。结论 该法准确、无干扰,可用于控制射干抗病毒注射液的质量。

OBJECTIVE To develope reversed-phase high performance liquid chromatographic method for the quantitative determinations of chlorogenic acid in Shegan Kangbingdu injection. METHODS The operation was carried out in the Kromasil ODS (5 μm, 4. 6 mm×200 mm) column made in Johnson Co. of Dalian with the mobile phase consisting of a mixture of water-methanol-acetonitril-acetic acid (95: 10: 5: 2, v/v), in which the flow rate of 1. 0 mL·min-1 and UV detection wavelength of 326 nm were set to determine the contents of the chlorogenic acid, at (23. 5± 0. 1)℃. RESULTS The linear range was 0. 0074 - 3. 7 μg (r=0. 9999), the relative standard deviations of the peak area for chlorogenic acid standard was 0. 60% and the standard was stable within 16 hours (RSD = 0. 54%). The average recovery was 100. 1%, and the detectable limit and quantitative limit were 2. 4 ng and 7. 4 ng. respectively. CONCLUSIONS The method is sensitive and accurate, and can be used for the quality control of the Shegan Kangbingdu injection.