该研究开发了一种基于HPLC自动进样系统的化学反应过程在线分析方法,以研究不同酸度的模拟柑橘汁中6’,7’-环氧佛手柑素(6’,7’-epoxybergamottin, 6’,7’-EB)的酸诱导反应产物和速率。编辑样品自动处理程序,使反应物在pH 2.0、3.5和5.0的25 ℃模拟果汁中充分反应,并优化色谱条件使反应物和主要产物完全分离,实现反应体系的周期性进样和分析;利用二极管阵列检测器和外标法对各物质进行定性和定量检测。6’,7’-EB在25 ℃酸催化下主要发生侧链环氧基开环水合反应,生成6’,7’-二羟佛手柑素,同时产生少量异欧前胡素,两反应均为一级反应;6’,7’-EB在pH 2.0和3.5条件下的半寿期分别为9.71和119.7 min,在pH 5.0时基本稳定。该方法实现了对6’,7’-EB水合反应速率的自动化在线分析,具有精度高和省时省力的特点,所得pH 3.5反应速率常数能较准确地预测葡萄柚汁中6’,7’-EB及其产物的浓度。
An on-line analysis method determining chemical reaction process based on an HPLC automatic sampling system was developed to study the products and reaction rates of 6′,7′-epoxybergamottin (6′,7′-EB) acid-induced reaction in simulated citrus juices with different acidity. The automatic pretreatment of imitated juice was programmed to ensure complete reaction at 25 ℃ and pH 2.0, 3.5 and 5.0, respectively; chromatographic conditions were optimized to separate the reactant and main products fully. Therefore, periodic sampling and analysis of these compounds in the reaction system was realized. Qualitative and quantitative detection of each substance was carried out by a diode array detector using the external standard method. At 25 ℃ by acid catalysis, 6′,7′-EB mainly involved in side-chain epoxide ring-opening and hydration reactions to generate 6′,7′-dihydroxybergamottin, with a few amounts of isoimperatorin produced too; both reactions followed the first-order kinetics. At pH 2.0 and 3.5, 6′,7′-EB showed a half-life of 9.71 and 119.7 min, respectively, whereas at pH 5.0 it was almost stable. This developed method achieved the automatic on-line analysis of 6′,7′-EB reaction rate, exhibiting the characteristics of both high precision and time/effort saving. The obtained reaction rate constant at pH 3.5 could be used to accurately predict the concentration of 6′,7′-EB and its products in real grapefruit juice.
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