高效液相色谱法同时测定发酵前后白参菌茶中六种活性成分

Simultaneous determination of changes in six active ingredients in Schizophyllum communeh fermented tea by HPLC

以绿茶和绿茶与白参菌共发酵茶为研究对象,建立快速同时检测绿茶与白参菌共发酵前后茶叶中的6种活性成分含量变化的高效液相色谱检测方法。将白参菌与绿茶共发酵,制备白参菌发酵茶。采用WondaSil C 18柱(4.6 mm×250 mm,5μm),乙腈(A)-0.2%H 3PO 4溶液(B)为流动相,梯度洗脱(0~15 min,5%A→15%A;15~40 min,15%A→25%A;40~45 min,25%A→5%A),流速1 mL/min,柱温30℃,检测波长为280 nm检测茶叶发酵前后的活性成分含量。结果表明,绿茶经过与白参菌共发酵后,咖啡碱和可可碱含量几乎没有变化,表没食子儿茶素、表儿茶素、表没食子儿茶素没食子酸酯、表儿茶素没食子酸酯的含量均大幅下降。6种活性成分在6.25~2000μg/mL下线性关系良好,相关系数(R)在0.9955~0.9999,检出限在0.3125~1.870μg/mL,绿茶的加标回收率在95.06%~111.06%,白参菌茶的加标回收率在85.06%~118.88%。白参菌与绿茶共发酵前后化学成分的含量发生变化,该液相检测方法快速、准确、重现性好,能够满足白参菌茶中多种活性成分同时定性、定量检测需求,具有一定的应用价值。

Using green tea and green tea co-fermented with Schizophyllum communeh(S.communeh fermented tea)as raw materials,we established a high-performance liquid chromatography(HPLC)-based method for rapid and simultaneous determination of changes in the contents of six active ingredients in green tea leaves before and after co-fermentation with S.communeh.Using a WondaSil C 18 column(4.6 mm×250 mm×5μm)with acetonitrile(A)and 0.2%phosphoric acid(B)as the mobile phase,we performed separation by gradient elution(0-15 min,5%A→15%A;15-40 min,15%A→25%A;40-45 min,25%A→5%A)with a flow rate of 1 mL/min and column temperature of 30℃.The contents of active ingredients in the tea leaves before and after fermentation were measured using a detection wavelength of 280 nm.The caffeine and theobromine contents in green tea barely changed after co-fermentation,whereas the epicatechin,epigallocatechin,epicatechin gallate and epigallocatechin gallate contents significantly decreased.All six active ingredients exhibited good linear relationships in the range of 6.25-2000μg/mL,with correlation coefficients in the range of 0.9955-0.9999.Detection limits were in the range of 0.3125-1.870μg/mL and the recovery rates obtained using the standard addition method were in the range of 95.06%-111.06%for green tea and 85.06%-118.88%for S.communeh fermented tea.The HPLC-based method developed in this study has potential applications in the testing of active components of tea products,as it is rapid and accurate,with good reproducibility,and has been proven suitable for simultaneous qualitative and quantitative determination of multiple active ingredients in S.communeh fermented tea.