HPLC法测定复方醋酸棉酚片中棉酚异构体含量

Determination of gossypol isomer in compound acetic acid gossypol tablets by HPLC

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 XDBC18色谱柱（250 mm×4.6 mm,5μm）,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1.88~37.6μg·mL-1和1.98~30.7μg·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 rain 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 KH2 pO4 buffer solution （ pH 3.0） -acetonitrile（20： 80）as the mobile phase at a flow rate of 1.0 mL · rain-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- t and 1.98 - 30. 7 μg · mL- l, respectively. The average recoveries were 101.0% and 101.2%, and RSDs were 2. 3 % and 2. 0% , respectively. Conclusion： The meth- od 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.