高效液相色谱法结合一测多评法对隔山消发酵前后3种苯乙酮类化合物含量变化的分析

采用高效液相色谱法结合一测多评法测定了隔山消发酵前后2,4-二羟基苯乙酮、2,5-二羟基苯乙酮和白首乌二苯酮等3种苯乙酮类化合物的含量。分别制备发酵前后的隔山消样品溶液,色谱分析中采用Thermo C18色谱柱作为固定相,用不同比例的水(A)、乙腈(B)和甲醇(C)的混合液作为流动相进行梯度洗脱,达到上述3种组分的良好分离。采用二极管阵列检测器在相同的检测波长(265nm处)分别测定3种组分的峰面积。分别用一测多评法(QAMS)和外标法计算所测定组分的含量,结果表明两种计算方法所得结果相似性极高,据此认为在隔山消发酵前后的样品溶液中各组分的质量评价中采用QAMS是可行的。按上述方法分析了同一批隔山消样品12份,取其中6份经发酵处理,对所得结果的平均值进行了比较,未经发酵的样品溶液中2,4-二羟基苯乙酮的质量分数为0.208mg·g^-1、2,5-二羟基苯乙酮的质量分数为0.152mg·g^-1,与经发酵的样品溶液中的质量分数(2,4-二羟基苯乙酮的质量分数为0.235mg·g^-1、2,5-二羟基苯乙酮的质量分数为0.180mg·g^-1)相比呈升高趋势,而白首乌二苯酮的质量分数由未经发酵样品溶液中的0.678mg·g^-1下降为发酵样品溶液中的0.543mg·g^-1。此外,研究结果还表明,不同含量的2,5-二羟基苯乙酮和白首乌二苯酮相对于2,4-二羟基苯乙酮(参照物)的相对校正因子和相对保留时间基本相同。

Analysis of Change of Contents of 3 Acetophenone Compounds in Cynanchum Bungei Decne Before and After Fermentation by HPLC with QAMS

HPLC with QAMS was applied to the determination of 3 acetophenone compounds,i.e.,2,4-dihydroxyacetophenone,2,5-dihydroxyacetophenone and baishouwu benzophenone in sample solutions of Cynanchum bungei Decne before and after fermentation.Sample solutions of Cynanchum bungei Decne before and after fermentation were prepared.In HPLC analysis,Thermo C18 chromatographic column was used as stationary phase and mixtures with various ratios of water(A),CH3 CN(B)and CH3 OH(C)were used as mobile phase in the gradient elution,and the 3 analytes mentioned above were well separated by this process,with their peak areas measured by DAD at the same wavelength of 265 nm.Contents of the 3 analytes were calculated separately by QAMS and external standard method.It was shown that results found by the 2 method were in high consistency and it was concluded that the use of QAMS in quality evaluation of the analytes in the sample solutions before and after fermentation of Cynanchum bungei Decne was feasible.12 portions of the same of Cynanchum bungei Decne from same batch were collected and testing solutions of 6 portions without fermentation and other 6 portions with fermentation were prepared and analyzed by the procedure described above.As shown by the results,average values of contents of 2,4-dihydroxyacetophenone and 2,5-dihydroxyacetophenone in solutions of the fermented samples were higher than those found in solutions of non-fermented samples(i.e.2,4-dihydroxyacetophenone raised from0.208 mg·g^-1 to 0.235 mg·g^-1,and 2,5-dihydroxyacetophenone from 0.152 mg·g^-1 to 0.180 mg·g^-1)and average content of baishouwu benzophenone in fermented samples was found lower than that in non-fermented samples(i.e.from 0.678 mg·g^-1 lowered to 0.543 mg·g^-1).In addition,it was shown that the values of relative correction factor and relative retention time found for 2,5-dihydroxyacetophenone and baishouwu benzophenone with different contents were same on the whole,taking 2,4-dihydroxyacetophenone as reference.