Simultaneous Determination of Carotenoids and Chlorophylls in Algae by High Performance Liquid Chromatography
高效液相色谱法测定藻类中的类胡萝卜素和叶绿素

An isocratic reversed-phase HPLC method for the simultaneous determination of carotenoids and chlorophylls in algae is presented. Both carotenoids and chlorophylls were separated by using a C18 column and a mobile phase consisting of dichloromethane/acetonitrile/methanol/water (22.5:9.5:67.5:0.5). The flow rate was 1.0mL/min. These pigments were detected by a UV-VIS absorbance detector set at 450nm. The average recoveries of these pigments by standard addition method were 99.1% for lutein, 98.5% for alpha-carotene, 99.4% for beta-carotene, 100.6% for chlorophyll a and 99.9% for chlorophyll b. The relative standard deviations of these pigments were 2.4% for lutein, 5.6% for alpha-carotene, 6.0% for beta-carotene, 4.1% for chlorophyll a and 4.0% for chlorophyll b. The calibration curves for these pigments gave good linearity over a wide range of 0-50mg/L (r = 0.9992-0.9999). The retention times were 3.0 min for lutein, 4.7 min for chlorophyll b, 6.3 min for chlorophyll a, 14.1 min for alpha-carotene, and 14.7 min for beta-carotene. Using this method, carotenoids and chlorophylls were determined in Spirulina platensis, Chlorella vulgaris and Haematococcus lacustris. Chlorella vulgaris and Haematococcus lacustris samples were ground in a B. Braun homogenizer. Spirulina platensis sample was smashed by supersonic wave. Acetone was used as extractant.