灰树花发酵过程天麻成分变化的HPLC检测方法研究

建立HPLC方法对天麻醇提取物成分及灰树花发酵液进行分析.色谱柱为Agilent TC-C18(4.6mm×250mm,5μm),流动相为A:0.1％磷酸水和B:乙腈,以梯度洗脱:0min～35min,B:3％～30％(V/V)；35min～45min,B:30％～70％ (Ⅴ/Ⅴ).流速1mL/min,柱温30℃,检测波长221nm.在此条件下,天麻提取液及灰树花发酵液中3种己确定的天麻成分具有良好的分离度,且线性关系良好；精密度高(RSD＜5％)；回收率在96％～102％之间,同时检测到经过6d发酵后发酵液中天麻素、对羟基苯甲醇和对羟基苯甲醛质量浓度与发酵第0d相比分别下降了46.38％、11.85％和36.75％.该方法可以用于灰树花发酵过程中天麻成分变化情况的跟踪检测,为进一步探明天麻成分与灰树花代谢相互作用打下基础,同时也初步探明了灰树花对于天麻成分的利用情况.

Determination of extracts from Gastrodia tuber fermented with Grifola frondosa by HPLC method

To establish a method for determination of gastrodin,p-hydroxybenzyl alcohol and p-hydroxybenzaldehyde in ethanol extracts fromGastrodia tuber and culture medium of G.frondosa.HPLC was used in the study.The compounds were eluted using a Agilent TC-C18（4.6mm×250mm,5μm） column,and detected at 221 nm with a flow rate of 1ml/min.The column tem-perature was set at 30℃.Gradient solvent A was 0.1% phosphoric acid;solvent B was acetonitrile,starting from 3% B to 30% B for 0~35min,30%~70% B for 35min~45min.Under the above condition,good linearity was obtained for three substances with a correlation coefficient 0.9999,recoveries ranged between 96% and 102%,while the precision of the method was 5%（RSD）.The content of three substances in culture medium of Grifola frondosa can be accurately determine simultaneously by this method.Lay the foundation for the further exploration of interaction ofG.frondosa metabolism and the ethanol extracts fromG.tuber.