RP-HPLC法测定羟苯苄酯有关物质的含量

Determination of the related substances of benzyl hydroxybenzoate by RP-HPLC

为了建立用于分析羟苯苄酯有关物质的RP-HPLC方法,分离条件为:Agilent ZORBAX Eclipse Plus Phenyl-Hexyl色谱柱,以甲醇-1%冰醋酸为流动相梯度洗脱,检测波长为254 nm,对其方法学进行了研究。结果显示,羟苯苄酯和对羟基苯甲酸的质量浓度分别在0.051~101.88μg/mL(r=1.00)和0.050~99.48μg/mL(r=0.999 8)范围内与峰面积呈良好线性关系,对羟基苯甲酸的平均加样回收率(%)为100.3,RSD(%)为0.95,对羟基苯甲酸定量限为0.24 ng。研究中还采用UPLC-Q-TOF对检出杂质结构进行了推测与鉴定。该研究为羟苯苄酯质量控制提供了理论依据。

An RP-HPLC method was established to separate the related substances of benzyl hydroxybenzoate. The separation was carried out on a Agilent ZORBAX Eclipse Plus Phenyl-Hexyl column. The mobile phase was methonal-0. 1% glacial acetic acid, using linear gradient elution, and the detection wavelength was 254 nm. There was a good linear relationship between 0. 051-101. 88 μg/mL（r=1. 00）and 0. 050-99. 48 μg/mL（r=0. 999 8）for benzyl hydroxybenzoate and p-hydroxybenzoic acid, repectively. The average recovery of p-hydroxybenzoic acid was 100. 3% and the RSD was 0. 95%. The LOQ of p-hydroxybenzoic acid was 0. 24 ng. The detected impurities were also identified by UPLC-Q-TOF. The established method is accurate and reproducible, and could be used for the quality control of benzyl hydroxybenzoate.