Nitao J K, Birr B A, Nair M G, Herms D A, Mattson W J
Department of Entomology, Michigan State University, East Lansing, Michigan 48824, USA.
J Agric Food Chem. 2001 May;49(5):2207-14. doi: 10.1021/jf001183b.
Proanthocyanidins (condensed tannins) frequently need to be quantified in large numbers of samples in food, plant, and environmental studies. An automated colorimetric method to quantify proanthocyanidins with sulfuric acid (H(2)SO(4)) was therefore developed for use in a continuous flow analyzer. Assay conditions were optimized using 50% methanol extracts of paper birch, sugar maple, and quaking aspen leaves. Short extraction times and centrifugation of samples prevented proanthocyanidin degradation that otherwise occurred in 50% methanol extracts of aspen leaves. Extraction of birch and maple proanthocyanidins with 50% methanol was comparable to or better than that with 70% acetone. Proanthocyanidin levels in aspen were lower when extracted with aqueous methanol, but relative differences among samples were consistent with those found in aqueous acetone extracts. Results from the automated sulfuric acid assay were highly correlated with those of the conventional BuOH-HCl method for proanthocyanidins and, except for birch, with the Folin--Denis assay for total phenolics. This new technique significantly improves assay processing rate and repeatability compared to conventional colorimetric proanthocyanidin assays.
在食品、植物和环境研究中,常常需要对大量样品中的原花青素(缩合单宁)进行定量分析。因此,开发了一种用硫酸(H₂SO₄)对原花青素进行定量的自动化比色法,用于连续流动分析仪。使用白桦、糖枫和颤杨树叶的50%甲醇提取物对测定条件进行了优化。较短的提取时间和样品离心操作可防止原花青素降解,否则颤杨树叶的50%甲醇提取物中会发生这种降解。用50%甲醇提取白桦和枫香的原花青素与用70%丙酮提取相当或更好。用水性甲醇提取时,颤杨中的原花青素含量较低,但样品之间的相对差异与水性丙酮提取物中的一致。自动化硫酸测定法的结果与传统的原花青素丁醇 - 盐酸法高度相关,除白桦外,与总酚的福林 - 丹尼斯测定法也高度相关。与传统的比色原花青素测定法相比,这项新技术显著提高了测定处理速度和重复性。