高效液相色谱法分析沙棘中类胡萝卜素的含量

Analysis of carotenoids content in sea buckthorn by high performance liquid chromatography

中文关键词:  沙棘  β-胡萝卜素  玉米黄质  β-隐黄质  叶黄素  高效液相色谱法

英文关键词:sea buckthorn  β-carotene  zeaxanthin  β-cryptoxanthin  lutein  high performance liquid chromatography

中文摘要:

      目的 建立沙棘中β-胡萝卜素、玉米黄质、β-隐黄质、叶黄素等4种类胡萝卜素的高效液相色谱分析方法, 并分析我国不同产地和不同品种沙棘中4种类胡萝卜素的赋存状态及含量差异, 以及不同采摘时期类胡萝卜素的变化规律。方法 沙棘鲜果经冷冻干燥预处理, 取适量沙棘粉进行皂化, 再用正己烷提取, 二氯甲烷-乙腈-甲醇(2:3:5, V:V:V)复溶, 经YMC Carotenoid 色谱柱(150 mm×4.6 mm, 3.0 μm)分离, 以二氯甲烷-乙腈-甲醇(2:3:5, V:V:V)为流动相, 等度洗脱, DAD检测器检测, 外标法定量。采集新疆、四川、吉林、山西和河北等地的19个沙棘样本, 并以新疆阿勒泰地区深秋红品种为代表, 从9月初至12月底分9期采样, 对样本中4种类胡萝卜素的含量进行分析。结果 β-胡萝卜素、玉米黄质、β-隐黄质、叶黄素等4种类胡萝卜素在1.0~200 mg/mL的浓度范围内具有良好的线性关系, 相关系数(r2)大于0.999, 方法检出限为2.0 mg/kg, 定量限为6.0 mg/kg。加标回收率为88.1%~92.7%, RSD为3.44%~6.45% (n=6)。19个沙棘样品中类胡萝卜素的总含量介于1468.4~6999.1 mg/kg之间, 平均含量为3287.9 mg/kg。β-胡萝卜素、玉米黄质、β-隐黄质、叶黄素的平均含量分别为581.3、1789.4、102.0、815.3 mg/kg。结论 本研究建立的分析方法准确、可靠, 对19个沙棘样本进行分析, 结果表明不同品种沙棘中4种类胡萝卜素的含量差异较大, 对不同采摘时期的样本进行分析, 结果表明11月中旬采摘的沙棘果类胡萝卜素的含量最高。

英文摘要:

      Objective To establish a high performance liquid chromatography (HPLC) method for simultaneous determination of 4 kinds of carotenoids (β-carotene, zeaxanthin, β-cryptoxanthin and lutein) in sea buckthorn, and to analyze the content difference of 4 kinds of carotenoids in different producing areas and different varieties of sea buckthorn in China, and to analyze the change rule of carotenoids in sea buckthorn at different picking periods. Methods The fresh berries of sea buckthorn were freeze-dried firstly. And then the sea buckthorn powder was extracted with hexane and reconstituted with dichloromethane acetonitrile methanol (2:3:5, V:V:V) after saponification. Then the sample was separated on a YMC Carotenoid column (150 mm×4.6 mm, 3.0 mm) with dichloromethane acetonitrile methanol (2:3:5, V:V:V) as mobile phase by isocratic elution. 4 kinds of carotenoids were detected by DAD detector and quantificated by external standard method. 19 samples were collected from Xinjiang, Sichuan, Jilin, Shanxi and Hebei Provinces. 9 samples of Shenqiuhong (from Xinjiang Autonomous Region) were collected from early September to the end of December. 4 carotenoids in these samples were analyzed by HPLC. Results β-carotene, zeaxanthin, β-cryptoxanthin and lutein had good linear relationships in the concentration range of 1.0-200 mg/mL, with correlation coefficients (r2) above 0.999. The limits of detection and limits of quantitation were 2.0 mg/kg and 6.0 mg/kg, respectively. The recoveries of 4 carotenoids ranged from 88.1%-92.7%, with relative standard deviations (RSD, n=6) of 3.44%?6.45%. The content of total carotenoids in 19 sea buckthorn samples were ranged from 1468.4 mg/kg to 6999.1 mg/kg, with an average content of 3287.9 mg/kg. The average content of β-carotene, zeaxanthin, β-cryptoxanthin and lutein was 581.3, 1789.4, 102.0, 815.3 mg/kg, respectively. Conclusion The established method of this study is accurate and reliable, it is found that the content of carotenoids in different subspecies of sea buckthorn is significantly different and the content of carotenoids in Shenqiuhong is highest in the mid of November.