HPLC-DAD同时测定复方尿维氨滴眼液中3种活性成分的含量

Simultaneous Determination of Three Active Ingredients in Compound Allantoin VitaminB6-E and Aminoethylsulfonic Acid Eye Drops by HPLC-DAD

目的应用高效液相色谱-二极管阵列检测器（HPLC-DAD）,建立复方尿维氨滴眼液中硫酸软骨素钠,尿囊素和维生素B6含量的同时测定方法。方法采用AlltimaC18色谱柱（4.6mm×250mm,5μm）,以25mmol·L-1磷酸二氢铵（含0.01%辛烷磺酸钠）-乙睛（95∶5）为流动相,流速0.5mL˙min-1,室温测定,硫酸软骨素钠,尿囊素和维生素B6的检测波长分别为195,215和291nm。结果硫酸软骨素钠,尿囊素和维生素B6的线性关系良好,线性相关系数（r）都大于0.9996;专属性强,3种成分和药品中其他成分以及强制降解产物之间都有良好的分离度;精密度和稳定性良好,峰面积的相对标准偏差（RSD）都小于2.0%;加样回收率良好,在99.01%~101.92%之间。结论该方法准确、简单、快速,可作为复方尿维氨滴眼液中硫酸软骨素钠,尿囊素和维生素B6的同时测定。

OBJECTIVE To establish a high performance liquid chromatography-diode array detection （HPLC-DAD） method for the simultaneous determination of chondroitin sulfate sodium, allantoin and vitamin B6 in Compound Allantoin VitaminB6-E and Aminoethylsulfonic Acid Eye Drops. METHODS An Alltima C18 column （4.6 mm×250 mm, 5 μm particle size） was used for the separation at room temperature, with 25 mmol·L^-1 ammonium dihydrogen phosphate （containing 0.01% beptanesulfonic acid sodium salt）-acetonitrile （95 ： 5） as the mobile phase at the flow rate of 0.5 mL·min^-1. The detection wavelengthes for chondroitin sulfate sodium, allantoin and vitamin B6 were 195, 215 and 291 nm, respectively. RESULTS The method showed good linearity for chondroitin sulfate sodium, allantoin and vitamin B6, with all correlation coefficients （r） greater than 0.999 6. The specificity study showed satisfactory resolutions between chondroitin sulfate sodium, allantoin, vitamin B6, other ingredients and forced degradation products. The precisions and stability were satisfactory with all relative standard deviations （RSD） of peak areas lower than 2.0%, and the spiked recovery of three ingredients ranged from 99.01% to 101.92%. CONCLUSION This method is an accurate, fast and simple method for the simultaneous determination of chondroitin sulfate sodium, allantoin and vitamin B6 in Compound Allantoin VitaminB6-E and Aminoethylsulfonic Acid Eye Drops.