黑曲霉固态发酵对淫羊藿苷含量的影响及检测

Effect of Solid State Fermentation of Herba Epimedii with Spergillus Niger on the Content of Icariin and Assay

目的:采用黑曲霉(Aspergillus niger)孢子悬液发酵淫羊藿,观察其对淫羊藿苷的影响,建立淫羊藿苷的检测方法。方法:淫羊藿在30℃、孢子浓度1.8×104/mL条件下固态预发酵7d。用水回流法、乙醇回流法、超声水提法、超声醇提法从淫羊藿中提取淫羊藿苷,HPLC法测定其含量。结论:发酵淫羊藿的色谱条件为:DiamonsilC18(250×4.6mm,5μm);流动相:甲醇-水-冰醋酸(60:40:0.25);检测波长:270nm;柱温:41℃;流速:1.0min/mL。发酵前后超声醇提法均优于其它方法,淫羊藿苷含量最高(5.06%、2.38%),水回流法(0.87、0.87%)、乙醇回流法(4.44、2.13%)、超声水提法(1.11、1.34%)发酵后淫羊藿苷含量发生变化,本研究为淫羊藿储藏期质量评价奠定基础。

Objective Herba Epimedii was fermented with spore suspension of Aspergillus niger and the method for detecting icariin during fermentation was established.Methods The solid-state fermentation condition was 30℃ for 7 days with spore density of 1.8×104/mL.The water reflux,ethanol reflux and ultrasonic water,ultrasonic ethanol extraction were applied for the extraction of icariin from Herba Epimedii.High performance liquid chromatography（HPLC） was employed to determine the icariin.Conclusion The suitable HPLC conditions for fermentation Herba Epimedii were the follows：column,diamonsil C18（250×4.6mm,5μm）;mobile phase,V（methanol）：V（water）：V（Acetic acid） = 60：40：0.25;detection wave length,270nm;temperature,41℃;flow rate of 1.0ml/min.For both unfermented and fermented Herba Epimedii,ultrasonic ethanol was the best among the above mentioned methods with high icariin content（5.06%,2.38%）,meanwhile,the water reflux,ethanol reflux and ultrasonic water extraction methods were（0.87,0.87%）,（4.44,2.13%）,and（1.11,1.34%）,respectively.This study has provided basis for the quality evaluation of Herba Epimedii storage.