摘要
建立了一种分离和测定雨生红球藻中虾青素和其它色素的反相高效液相色谱法 ;含有雨生红球藻细胞的培养液经离心分离 ,匀浆破碎 ,用甲醇 -二氯甲烷 (体积比3∶1)提取色素 ,提取液用HPLC分离测定 ;分析柱为Nova_PakC18(300mm×3.9mmID ,5μm)柱 ,流动相为水、甲醇和丙酮 ,流速1.0mL/min ,用二极管阵列检测器在250～700nm波长范围扫描 ,在476nm处进行检测 ;虾青素、多种类胡萝卜素和叶绿素在30min内得到较好分离 ,其中游离虾青素、斑蝥黄质和 β_胡萝卜素的检出限分别为3.56、1.36和29.4μg/L,这3种类胡萝卜素分别在质量浓度为1.99～31.7mg/L、2.44～39.0mg/L、2.25～36.0mg/L范围内与峰面积线性关系良好 ,相关系数分别为0.9993、0.9987和0.9778;该法的精密度 (RSD为0.09 %～2.97% )和回收率(96%～102 % )均符合方法学要求 。
A gradient reversed_phase high performance liquid chromatography(RP_HPLC)was developed for the separation of astaxanthin and other pigments from the pigment extracts of the microalga Haematococcus pluvialis.Algal cells were collected by centrifuging the culture fluid at3000g for15min,homogenized and extracted using the solvent mixture of methanol and dichloromethane(3∶1by volume).Chromatographic separation was performed in Nova_Pak C 18 column(300mm×3.9mm ID,5μm)at25℃with an eluent consisting of water,methanol and acetone at a flow rate of1.0mL/min,the eluate was monitored by a photodiode array detector at476nm.Sepˉaration and identification of chlorophylls and multiple carotenoids were completed in30min.The detection limits of astaxanthin,canthaxanthin andβ_carotene were3.56,1.36and29.4μg/L,respectively.The linear ranges of astaxanthin,canthaxanthin andβ_carotene were1.99～31.7mg/L(r=0.9993),2.44～39.0mg/L(r=0.9987)and2.25～36.0mg/L(r=0.9778),respectively.This method is rapid,simple,precise and accuˉrate(RSD0.09%～2.97%,average recoveries96%～102%),and can be used for the quantification of astaxˉanthin and other pigments in Haematococcus Pluvialis.
出处
《分析测试学报》
CAS
CSCD
北大核心
2003年第4期28-31,共4页
Journal of Instrumental Analysis
基金
"十五"国家科技攻关项目 (2002BA514A -10)
