Abstract: To establish an optimized method for the determination of berberine hydrochloride in the crystals of Phellodendron chinensis by high performance liquid chromatography (HPLC), and to evaluate the inhibitory activity of P. chinense against drug-resistant bacteria, a new method was determined by optimizing the mobile phase and the detection conditions, and the antibacterial activity of the drug-resistant strain was evaluated by the antibacterial test in vitro. The results showed that the mobile phase was consisted of acetonitrile-0.1% phosphoric acid solution (50∶50) (0.1 g sodium dodecyl sulfate per 100 mL phosphoric acid solution), the detection wavelength was 265 nm, the flow rate was 1 mL·min-1, column temperature was 30 ℃. When the concentration of berberine hydrochloride was between 103 mg·L-1 and 515 mg·L-1, there was a good linear relationship between berberine and their corresponding peak area integral values (R2 ≤ 0.999 7). Under these conditions, the precision, stability and repeatability of the determination process were tested with RSD of 0.49%, 1.23% and 0.29%, respectively, which indicated that the test method was accurate. The average recovery rate was 97.84%～98.06%, and the RSD was 0.44%～1.00%, which indicated that the method was accurate and effective. The bacteriostatic activity test showed that the crystals of P. chinense had a good inhibitory effect on drug-resistant Staphylococcus aureus and Pseudomonas aeruginosa. The optimized method was safe, rapid, accurate and effective, which could be used for analysis and determination of berberine hydrochloride in P. chinensis, and berberine hydrochloride had a good bacteriostatic effect. The research broadened the types of natural products of resistant bacteria and provided a basis for clinical and research of berberine hydrochloride.

Key words: Phellodendron chinensis crystal, berberine hydrochloride, high performance liquid chromatography, bacteriostasis activity

摘要： 为建立高效液相色谱（HPLC）测定川黄柏结晶物中盐酸小檗碱含量的优化方法，并评价川黄柏对耐药性细菌的抑制活性，对川黄柏提取物进行结晶处理后，通过对流动相及检测条件的优化确定最优检测方法；用结晶物对耐药性菌株进行体外抑菌试验评价抑制活性。结果表明，检测最佳条件：流动相为乙腈-0.1%磷酸溶液（50∶50）（0.1 g十二烷基硫酸钠每100 mL磷酸溶液），检测波长为265 nm，流速1 mL·min-1，柱温30 ℃；当盐酸小檗碱浓度在103~515 mg·L-1之间时，与各自对应峰面积积分值呈较好的线性关系（R2=0.999 7）；对该条件下测定过程进行精密度、稳定性、重复性试验检测，RSD分别为0.49%、1.23%、0.29%，表明试验方法准确；平均加样回收率为97.84%~98.06%，RSD为0.44%~1.00%，说明检测方法准确有效。抑菌活性试验发现川黄柏结晶物对耐药性金黄色葡萄球菌(Staphylococcus aureus)和铜绿假单胞菌(Pseudomonas aeruginosa)均有很好的抑制效果。优化方法安全、快速、精确、有效，可用于黄柏药材中盐酸小檗碱的分析测定；川黄柏中盐酸小檗碱具有很好地抑菌作用。研究拓宽了对抗耐药性细菌的天然产物种类，为其临床应用和科学研究奠定基础。

关键词: 黄柏结晶物, 盐酸小檗碱, 高效液相色谱, 抑菌活性