海娜花抗真菌有效成分的提取工艺优化与抑菌效力研究

目的 研究海娜花抗真菌有效成分的提取工艺优化与抑菌效力。方法 分别采用回流提取法与超声提取法提取海娜花。采用高效液相色谱作定量分析,流动相A为磷酸二氢铵30 mmol/L、B为甲醇（用磷酸调pH=3）,梯度洗脱,温度为30℃,进样量为20μL,色谱柱为Agilent Zorbax SB-C18柱（250 mm×4.6 mm,5μm）。考察溶剂、温度、时间对有效成分提取效果的影响,确定最佳提取工艺。采用美国临床实验室标准化委员会（NCCLS）产孢丝状真菌抗真菌药敏试验方案M38-A,以白色念珠菌为模型菌,对海娜花提取物的抗真菌效力进行初步研究。结果与结论 回流提取法的最佳工艺条件为10倍量70%乙醇溶液,100℃下回流提取1.0 h,共提取3次。超声提取法的最佳工艺条件为10倍量70%乙醇溶液,50℃下超声提取1.5 h,共提取3次。海娜花提取物对白色念珠菌的最低抑菌浓度（MIC）为320～640μg/mL。

Research of Extraction Technology Optimization of Antifungal Effective Constituents from Lawsonia Inermis L. and Its Antifungal Potency

Objective To study the extraction technology optimization of antifungal constituents from Lawsonia inermis L. and its antifungal potency. Methods The reflux extraction method and the ultrasonic extraction method were adopted to extract the Lawsonia inermis L. The high performance liquid chromalography（HPLC） method was adopted to perform the quantitative analysis. The mobile phase A was 30 mmol/L ammonium dihydrogen phosphate and the mobile B phase was methanol（adjusting to pH =3 with phosphoric acid） with the gradient elution. The temperature was 30 ℃, the injection volume was 20 μL. The chromatographic column was the Agilent Zorbax SB-Cts column （250 mm ×4.6 mm,5μm）. The influence of solvent,temperature and time on the extraction effect of effective ingredients was investigated. The optimum extraction process was determined. The spores- producing filamentous fungi antifungal susceptibility testing scheme M38-A proposed by the American Committee for clinical laboratory standards（NCCLS） was adopted. With Candida albicans as the model organism,the antifungal potency of Lawsonia inermis L. extract was preliminarily studied. Results and Conclusion The optimal technological condition of the reflux extract method was 10 times the amount of 70% ethanol solution,reflux extraction for 1 h at 100 ℃ and extracting for 3 times. The optimal technological condition of the ultrasonic extraction method was 10 times the amount of 70% ethanol solution, ultrasonic extraction for 1.5 h at 50 ℃ and extracting for 3 times. The minimal inhibitory concentration（MIC） of Lawsonia inermis L. extract on Candida albicans was 520 -640 μg/mL.