高效液相色谱法测定缓解视疲劳片中原花青素的含量

Determination of Proanthocyanidins in Asthenopia-relieving Tablets by HPLC

目的建立缓解视疲劳片中原花青素的含量测定方法。方法原花青素经铁盐催化降解为花青素离子后进行HPLC分析。通过正交试验优化降解条件为:1.0 mL供试品溶液加10 mg/mL硫酸高铁铵溶液0.2 mL和正丁醇-盐酸混合液(97:3,v/v)8.8 mL,100℃水浴加热1 h。色谱条件:Ultimate XB-C18色谱柱(4.6 mm×150 mm,5μm),流动相为甲醇-0.1%磷酸(45:55),流速0.5 mL/min,柱温30℃,检测波长530 nm。结果原花青素在考察的浓度范围内线性良好(r=0.9994),检出限9.33μg/mL,定量限31.12μg/mL,平均回收率95.56%。结论此方法操作简单,重现性好,可为该产品的质量控制提供参考。

Objective To establish a method for the determination ofproanthocyanidins in Asthenopia-relieving Tablets by HPLC.Methods The content of proanthocyanidins was determined after degradation in n-butanol-HC1 reaction medium using iron salt as catalyst.The degradation condition was optimized through single factor and orthogonal experiments,which was that taking 0.2 mL 10 g/L ferric ammoniun sulfate solution and 8.8 mL n-butanol-HCl（93∶7,v/v） mixture into 1 mL sample solution and reacting at 100 ℃ for 1 h.The analysis was achieved on the Ultimate XB-C18 （4.6 mm× 150 mm,5 μm） column,with mobile phase consisted of methanol and 0.1％ phosphoric acid（45∶55,v/v） at a flow-rate of 0,5 mL/min.The column temperature was set at 30 ℃ and wavelength of detector was 530nm.Results The calibration curve showed good linear regression,the detection limit was 9.33 μg/mL and the limit of quantitation was 31.12 μg/mL.The average recovery of assay was 95.56 ％.Conclusion The method is full of simplicity,reproducibility and accuracy.It can offer a reference for the quality evaluation and control of this product.