HPLC法同时测定双金连合剂中11种成分的含量

目的: 建立反相高效液相色谱法同时测定双金连合剂中新绿原酸、绿原酸、隐绿原酸、连翘苷、芦丁、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、牡荆素和黄芩素的含量。方法: 采用Phenomenex ODS C₁₈ 色谱柱(4.6 mm×250 mm,5 μm),以乙腈(A)-0.4%磷酸(B)为流动相,梯度洗脱(0~10 min,5%A→9%A;10~30 min,9%A;30~60 min,9%A→30%A;60~90 min,30%A→50%A),流速1.0 mL · min^-1,最大吸收波长下检测(新绿原酸、绿原酸和隐绿原酸:327 nm;异绿原酸B、A、C:330 nm;芦丁、黄芩苷和黄芩素:280 nm;连翘苷:277 nm;牡荆素:350 nm)。结果: 11种待测成分质量浓度与峰面积在测定范围内均呈良好线性关系(r≥0.999 0),平均回收率为96.7%~101.6%(RSD≤2.8%)。结论: 该方法经方法学验证可为更好地控制双金连合剂的质量提供参考依据。

Objective: To establish an HPLC method for simultaneous determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, forsythin, rutin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, baicalin, vitexin, and baicalein in Shuangjinlian mixture. Methods: The chromatographic separations were obtained on a Phenomenex ODS C₁₈ column(4.6 mm×250 mm, 5 μm), with the mobile phase consisting of acetonitrile(A)-0.4% phosphoric acid(B)in gradient elution mode(0-10 min, 5%A→9%A;10-30 min, 9%A;30-60 min, 9%A→3 0%A;60-90 min, 30%A→50%A), at the flow rate of 1.0 mL · min^-1.The UV detection wavelength was set at the maximum absorption wavelengths, 327 nm for neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid, 330 nm for isochlorogenic acids B, A, and C, 280 nm for rutin, baicalin and baicalein, 277 nm for forsythin, and 350 nm for vitexin. Results: The good linearity between the concentration and peak area was obtained for the 11 components in the determined ranges(r≥0.999 0)with the average recovery of 96.7%-101.6%(RSD≤2.8%). Conclusion: Methodology validation showed that the method can be used for the quality control of Shuangjinlian mixture.