反相高效液相色谱法测定果蔬中的β-胡萝卜素

Determination of β-carotene in fruits and vegetables by reversed-phase high performance liquid chromatography

从检测波长、流动相、柱温等方面优化了反相高效液相色谱测定果蔬中的β-胡萝卜素的色谱条件,并对方法的应用性进行了评价。用单因子实验方法优化得到的反相高效液相色谱分析果蔬中β-胡萝卜素的条件为:采用1:1 (V/V)丙酮-石油醚混合液直接提取,提取液减压蒸干后,用丙酮溶解,经NOVA-PAK(150mm×3.9mm,4μ, Waters)C18色谱柱,以乙腈-甲醇(20:80)为流动相进行洗脱,用2487紫外检测器在450nm下进行检测。在此条件下,β-胡萝卜素标准曲线的回归方程为Y=45739X- 19784,相关系数大于0.999,线性范围为0-50mg/L,最小检出限为0.10mg/L,加标样品回收率为91.7%-99.7%,变异系数CV(%)为4.4%-5.0%。该方法能简便、快速、准确地测定果蔬样品中的β-胡萝卜素。

Wavelength, mobile phase and column temperature were optimized for developing a reversed -phase high performance liquid chromatographic method for determination of β-carotene in fruits and vegetables. Also, the validity of the proposed method was studied. Carotenoids were extracted with acetone and petroleum ether(1:1) ,the extract was dried under vacuum condition and further dissolved in acetone, then,separated on a NOVA-PAK reverse phase C18 column (150mm×3.9mm, 4μ, Waters Inc) with the mobile phase composed of acetonitrile and methanol at ratio of 20 to 80. The separation was monitored at 450nm using a 2487 UV Detector. Under the optimal condition, when concentration of β -carotene range from 0mg/L to 50mg/L,the regressive equation of the calibration graph was y= 45739X-19784, R square value was higher than 0.999, indicating the equation has good linearity.The detection limit was 0.10mg/L, recovery rate was 91.7% -99.7% , coefficients of variation were 4.4%-5.0%, which all suggested the proposed method was simple,rapid and accurate to be used in determination of β-carotene in fruits and vegetables.