采用尼罗红荧光探针对微藻中油脂的定量测定  

Quantification of lipid content in microalgae using Nile red as fluorescent probe

作　　者：梁文艳[1] 张元春[1] 曹敬灿[1] 汪丽[1] 于建[1] 

机构地区：[1]北京林业大学环境科学与工程学院,北京100083

出　　处：《环境化学》2013年第8期1491-1495,共5页Environmental Chemistry

基　　金：中央高校基本科研业务费专项(BLYX200909);国家水体污染控制与治理科技重大专项(2012ZX07307-001-006)资助

摘　　要：以尼罗红为荧光探针,对小球藻油脂含量的定量分析条件进行了研究.测定中,使用48孔酶标仪作为荧光检测器.研究显示,当荧光激发光波长为485 nm,发射光波长为580 nm时,适用于测定微藻中的中性油脂含量.当二甲基亚砜终浓度为20%,尼罗红终浓度为0.5μg.mL-1,藻液与尼罗红置于40℃暗处反应5 min后,能够获得最大的相对荧光测定值.当藻液光密度OD680在0.61至1.73时,藻液油脂含量与尼罗红荧光测定值呈现良好的线性关系,相关系数R2为0.9836,可以通过尼罗红荧光值的测定推算油脂的含量.This paper studied the quantification of the lipid content of Chlorella vulgaris by Nile red assay.Fluorescence could measured on a 48-well plate spectrofluorometer.The results showed that the excitation wavelength of 485 nm and the emission wavelength of 580 nm were optimal for the detection of neutral lipid.The maximum fluorescence could be reached when the final concentration of dimethyl sulfoxide was 20%,and the final concentration of Nile red was 0.5 μg.mL-1,after 5 min reaction of microalgae with NR at 40 ℃.When the optical density OD680 of algal solution was between 0.61 and 1.73,the fluorescence of Nile red assay had a linear relationship with the lipid content of algae with the R2 of 0.9836.It is concluded that the nile red assay can be used to detect the lipid content of Chlorella vulgaris.

关 键 词：尼罗红 油脂 微藻 定量 

分 类 号：Q949.93[生物学—植物学] O657.3[理学—分析化学]