四种原花青素含量测定方法比较

本文对四种原花青素(PC)定量分析方法包括:硫酸-香草醛法、钼酸铵法、正丁醇-盐酸-HPLC法和硫解-HPLC法进行了评价。结果表明,线性相关系数均大于≥0.999,LOD分别为:2.21、5.47 μg/mL、7.3 ng和≤2.0 ng,RSD分别为:2.52%、2.42%、1.85%和≤0.70%,加标回收率分别为:94.24%、96.20%、98.10%和≥40.81%。硫酸-香草醛法的显色反应液不够稳定,其余3种方法10 h内较稳定。用这4种方法对市售的葡萄籽、葡萄皮、花生衣及松树皮的PC提取物共计21份样品进行了含量分析,各方法测定结果分别为:38.70%、18.80%、40.24%和8.39%。硫酸-香草醛法与HPLC法检测结果间的相关系数最高,为0.5311,其次HPLC法与硫解-HPLC法之间的相关系数为0.4001,其余相关系数均小于0.2。采用硫解-HPLC法分析21份样品PC的平均聚合度在1.21~5.04之间。通过方差分析,可得知方法、样品以及两者的交互作用都对原花青素含量的影响达到了极显著的水平。

Comparison of Four Methods for Quantitation of Proanthocyanidin

Four methods for quantitation of proanthocyanidins(PC)including:sulfuric acid-vanillin method,ammonium molybdate method,N-butanol hydrochloric acid-HPLC method and thiolysis-HPLC method were evaluated in this paper. The results showed that linear correlation coefficients of the four methods were greater than 0.999,and the minimum detection limits were:2.21,5.47 μg/mL,7.3 ng and ≤2.0 ng,RSD were:2.52%,2.42%,1.85% and ≤0.70%,and the recoveries were:94.24%,96.20%,98.10% and ≥40.81%.The chromogenic reaction solution of the sulphuric acid vanillin method was not stable enough,and the other three methods were relatively stable within 10 h. A total of 21 samples of PC extract from commercially available grape seed,grape skin,peanut clothing and pine bark were analyzed by the methods. The results were 38.70%,18.80%,40.24% and 8.39%,respectively. The correlation coefficient between the results of sulfuric acid-vanillin method and HPLC method was the highest,0.5311,and the correlation coefficient between HPLC method and thiolysis-HPLC method was 0.4001,and the other correlation coefficients were less than 0.2.The average degree of polymerization of 21 samples of PC was analyzed by thiolysis-HPLC method between 1.21 and 5.04.Through analysis of variance,it could be seen that method,sample and the interaction between method and sample had a very significant effect on the proanthocyanidin content.