Dzakovich Michael P, Gas-Pascual Elisabet, Orchard Caleb J, Sari Eka N, Riedl Ken M, Schwartz Steven J, Francis David M, Cooperstone Jessica L
The Ohio State University, Department of Horticulture and Crop Science, 2001 Fyffe Court, Columbus, OH 43210.
The Ohio State University, Ohio Agricultural Research and Development Center, Department of Horticulture and Crop Science, 1680 Madison Ave, Wooster, OH 44691.
J AOAC Int. 2019 Jul 1;102(4):1069-1079. doi: 10.5740/jaoacint.19-0017. Epub 2019 Feb 20.
Tomatoes () are an economically and nutritionally important crop colored by carotenoids such as lycopene and β-carotene. Market diversification and interest in the health benefits of carotenoids has created the desire in plant, food, and nutritional scientists for improved extraction and quantification protocols that avoid the analytical bottlenecks caused by current methods. Our objective was to compare standard and rapid extraction as well as chromatographic separation methods for tomato carotenoids. Comparison was based on accuracy and the ability to discriminate between alleles and genetic backgrounds. Estimates of the contribution to variance in the presence of genetic and environmental effects were further used for comparison. Selections of cherry and processing tomatoes with varying carotenoid profiles were assessed using both established extraction and HPLC-diode array detector (HPLC-DAD) methods and rapid extraction and ultra-HPLC-DAD (UHPLC-DAD) protocols. Discrimination of alleles in samples extracted rapidly (<5 min/sample) was similar to samples extracted using a standard method (10 min/sample), although carotenoid concentrations were lower due to reduced extraction efficiency. Quantification by HPLC-DAD (21.5 min/sample) and UHPLC-DAD (4.2 min/sample) were comparable, but the UHPLC-DAD method could not separate all carotenoids and isomers of tangerine tomatoes. Random effects modeling indicated that extraction and chromatographic methods explained a small proportion of variance compared with genetic and environmental sources. The rapid extraction and UHPLC-DAD methods could enhance throughput for some applications compared with standard protocols.
番茄（）是一种在经济和营养方面都很重要的作物，其颜色由类胡萝卜素如番茄红素和β-胡萝卜素赋予。市场的多样化以及对类胡萝卜素健康益处的关注，使得植物、食品和营养科学家们渴望改进提取和定量方案，以避免当前方法造成的分析瓶颈。我们的目标是比较番茄类胡萝卜素的标准提取法和快速提取法以及色谱分离方法。比较基于准确性以及区分等位基因和遗传背景的能力。在存在遗传和环境效应的情况下，对变异贡献的估计进一步用于比较。使用既定的提取方法和高效液相色谱 - 二极管阵列检测器（HPLC - DAD）方法以及快速提取法和超高效液相色谱 - 二极管阵列检测器（UHPLC - DAD）方案，对具有不同类胡萝卜素谱的樱桃番茄和加工番茄进行了评估。快速提取（<5分钟/样品）的样品中等位基因的区分与使用标准方法（10分钟/样品）提取的样品相似，尽管由于提取效率降低，类胡萝卜素浓度较低。HPLC - DAD（21.5分钟/样品）和UHPLC - DAD（4.2分钟/样品）的定量结果相当，但UHPLC - DAD方法无法分离所有类胡萝卜素和橘黄色番茄的异构体。随机效应模型表明，与遗传和环境来源相比，提取和色谱方法解释的变异比例较小。与标准方案相比，快速提取和UHPLC - DAD方法在某些应用中可以提高通量。
