食品中叶黄素、玉米黄质和β-隐黄质同时检测方法的建立

The Establishment of a Simultaneous Detection Method for Lutein,Zeaxanthin and β-Cryptoxanthin in Food

本文采用高效液相色谱法建立了食品中叶黄素、玉米黄质、β-隐黄质同时检测的方法,最终确定最优前处理条件为:试样中加入30 mL含0.1%的2,6-二叔丁基-4-甲基苯酚(BHT)乙醇溶液和10 mL 60%氢氧化钾水溶液,置于恒温水浴箱内50℃振荡皂化15 min,处理样液用100 mL正己烷∶乙醚∶环己烷(2∶2∶1)混合液萃取。结果显示,实验建立的方法提取效果优于国标方法和超声提取方法,可以实现叶黄素的快速温和提取,提取效果较佳。以小米和大米为样品进行方法学验证,结果显示,叶黄素、玉米黄质、β-隐黄质分别在0.2342~9.3689、0.3004~12.0167、0.3734~14.9371μg/mL范围内线性关系良好,平行测定6次,叶黄素、玉米黄质的相对标准偏差(RSD)分别为1.41%、4.98%,小米中未检测到β-隐黄质,加标回收率均在85.6%~98.8%,表明该方法重现行良好、准确可行。该方法的建立可以弥补相关标准的不足,解决实际工作中存在的问题,为产品开发,食物评价和监管提供技术方法。

In this study,a method for the simultaneous analysis and detection of lutein,zeaxanthin and β-cryptoxanthin in food was developed using high-performance liquid chromatography(HPLC). The optimal pretreatment conditions were adding with 30 mL 0.1% 2,6-di-tertbutyl-4-methyl phenol(BHT)ethanol solution and 10 mL 60% potassium hydroxide solution for samples. Then the samples were saponificated in a water bath for 15 min at a constant temperature of 50 ℃ by thermostatic waterbath,followed by extraction using 100 mL solvent mixture of n-hexane,ethyl ether and cyclohexane(2∶2∶1). The results showed that this method was superior to the national standard and ultrasonic extraction,because it could be applied rapidly and the mild extraction of lutein provided an enhanced extraction effect. A millet and rice sample was used for methodological verification and the results showed that the linear relations for lutein,zeaxanthin and β-cryptoxanthin were good in the range of 0.2342~9.3689,0.3004~12.0167 and 0.3734~14.9371 μg/mL,respectively. Six parallel experiments were conducted and the relative standard deviations of lutein and zeaxanthin were 1.41% and 4.98%,respectively,and their spiked recoveries were between 85.6% and 98.8%,thus indicating that this method had sound repeatability and reliability characteristics. The establishment of this method can make up for the current deficiency of relevant standards. Furthermore,it provides solutions to various problems in practical work,as well as technical assistant for product development,food evaluation and supervision in the food production industry.