Development, validation and evaluation of an analytical method for the determination of monomeric and oligomeric procyanidins in apple extracts
Development, validation and evaluation of an analytical method for the determination of monomeric and oligomeric procyanidins in apple extracts
复制标题
DOI：
10.1016/j.chroma.2017.03.030
复制
发表时间：
2017-04-28
影响因子：
4.1
通讯作者：
Kroon, Paul A.
中科院分区：
文献类型：
--
作者：
Hollands, Wendy J.;Voorspoels, Stefan;Kroon, Paul A.
苹果和苹果提取物中个别低聚原花青素的数据缺乏。我们的目的是开发，验证和评估的分析方法的分离，鉴定和定量的单体和低聚黄烷醇在苹果提取物。为了实现这一点，我们从冻干苹果中制备了两种类型的黄烷醇提取物;一种是富含表儿茶素的提取物，其含有类似于30%（w/w）单体（-）-表儿茶素，其还含有低聚原花青素（提取物A），第二种是富含低聚原花青素的提取物，其耗尽了表儿茶素（提取物B）。方法优化考虑的参数为HPLC色谱柱和条件、样品加热、提取物质量和稀释体积。方法验证考虑的性能特征包括标准品线性、方法灵敏度、精密度和准确度。8个实验室参与了方法评价。分析物的色谱分离最好利用Hilic柱与二元移动的相组成的酸性乙腈和酸性含水甲醇。最终方法显示表儿茶素在5-100 μ g/mL范围内呈线性，相关系数>0.999。分析物的日内和日间精密度范围分别为2 - 6%和2 - 13%。直到dp 3，该方法的正确率>95%，但随着dp的增加而下降。实验室内精密度显示RSD值
There is a lack of data for individual oligomeric procyanidins in apples and apple extracts. Our aim was to develop, validate and evaluate an analytical method for the separation, identification and quantification of monomeric and oligomeric fiavanols in apple extracts. To achieve this, we prepared two types of flavanol extracts from freeze-dried apples; one was an epicatechin-rich extract containing similar to 30% (w/w) monomeric (-)-epicatechin which also contained oligomeric procyanidins (Extract A), the second was an oligomeric procyanidin-rich extract depleted of epicatechin (Extract B). The parameters considered for method optimisation were HPLC columns and conditions, sample heating, mass of extract and dilution volumes. The performance characteristics considered for method validation included standard linearity, method sensitivity, precision and trueness. Eight laboratories participated in the method evaluation. Chromatographic separation of the analytes was best achieved utilizing a Hilic column with a binary mobile phase consisting of acidic acetonitrile and acidic aqueous methanol. The final method showed linearity for epicatechin in the range 5-100 mu g/mL with a correlation co-efficient >0.999. Intra-day and inter-day precision of the analytes ranged from 2 to 6% and 2 to 13% respectively. Up to dp3, trueness of the method was >95% but decreased with increasing dp. Within laboratory precision showed RSD values