HPLC法测定复方醋酸棉酚片中棉酚异构体含量

目的: 建立测定复方醋酸棉酚片中(-)-棉酚及(+)-棉酚含量的高效液相色谱方法。方法: 以N,N-二甲基甲酰胺为溶剂,(R)-(-)-2-胺基-1-丙醇为衍生化试剂,90 ℃下水浴加热30 min进行衍生化反应。含量测定使用Agilent XDB-C₁₈色谱柱(250 mm×4.6 mm,5 μm),流动相为10 mmol·L^-1 磷酸二氢钾缓冲液(pH=3.0)-乙腈(20:80),流速1.0 mL·min^-1,检测波长247 nm,柱温40 ℃。结果: (-)-棉酚和(+)-棉酚浓度分别在1.88~37.6 μg·mL^-1和1.98~30.7 μg·mL^-1范围内呈良好线性关系;(-)-棉酚及(+)-棉酚的平均回收率分别为101.0%和101.2%,RSD分别为2.3%和2.0%。结论: 该方法可以准确测定复方醋酸棉酚片中(-)-棉酚及(+)-棉酚含量,有效提高其现有标准的质量控制水平。

Objective: To establish an HPLC method for quantitative determination of (-)-gossypol and (+)-gossypol in compound acetic acid gossypol tablets. Methods: Derivatization was conducted with (R)-(-)-2-Amino-1-propanol as the derivatization reagent and the sample was heated in a water bath at 90 ℃ for 30 min with N,N-dimethylformamide as the solvent.The content of (-)-gossypol and (+)-gossypol was detected by HPLC in an Agilent XDB-C₁₈ column(250 mm×4.6 mm,5 μm)with 10 mmol·L^-1 KH₂PO₄ buffer solution(pH 3.0)-acetonitrile(20:80)as the mobile phase at a flow rate of 1.0 mL·min^-1.The detection wavelength was 247 nm and the column temperature was maintained at 40 ℃. Results: There were good linear relationships for (-)-gossypol and (+)-gossypol in the ranges of 1.88-37.6 μg·mL^-1 and 1.98-30.7 μg·mL^-1,respectively.The average recoveries were 101.0% and 101.2%,and RSDs were 2.3% and 2.0%,respectively. Conclusion: The method is reliable for the determination of(-)-gossypol and(+)-gossypol in compound acetic acid gossypol tablets and can effectively improve the quality control level of the existing standard.