快速、可靠、简便的总脂含量测定方法在富油藻种筛选、功能性藻油评估及微藻生物学研究中至关重要。本研究以三种微藻（微绿球藻Nannochloris sp.、普通小球藻Chlorella vulgaris、绿色巴夫藻Pavlova viridis）藻粉为原料，系统比较了四种方法（Bligh-Dyer法、改良Bligh-Dyer法、ASTM标准法和皂化法）用于测定微藻中总脂含量的差异，并以原位转酯法为对照，进一步分析了各方法总脂提取物中的脂肪酸含量及组成。改良Bligh-Dyer法和ASTM标准法总脂测定结果稳定，脂肪酸回收率最高，三种藻粉中分别为93.65%和93.74%、85.02%和85.77%、88.62%和88.84%；两者总脂含量及提取物中脂肪酸的组成和含量均无显著差异（p>0.05）。显然，改良Bligh-Dyer法操作简便、分析时间短（1 h）、所需样品量少（0.25 g），可替代ASTM标准法，作为实验室快速准确测定微藻总脂含量的首选方法。

The rapid, reliable and easy manipulated method for determination of total lipid content in microalgae is crucial important for screening oleaginous strains, evaluation of functional oil from microalgae and microalgal biological research. Using dry powder of three microalgae (Nannochloris sp., Chlorella vulgaris and Pavlova viridis) as feedstock, the difference of four methods (Bligh-Dyer, modified Bligh-Dyer, ASTM standard and direct saponification methods) was systematically compared in terms of determining total lipid content in microalgae. The fatty acid contents and compositions in crude extracts from different methods were further profiled by GC/MS with in suit transesterification method as control. The results showed that the modified Bligh-Dyer and ASTM standard methods had steady extraction outcomes with the highest fatty acid recoveries at 93.65% vs 93.74%, 85.02% vs 85.77%, and 88.62% vs 88.84% in three powders, respectively. Moreover, no significant difference (p>0.05) appeared in total lipid and fatty acid contents and compositions in extracts by the two methods. Obviously, the modified Bligh-Dyer method is a simple and preferred method with less amount demand of sample (0.25 g) and short time-consuming (1.0 h) for the rapid determination of total lipid content in microalgae, which is an alternative of ASTM standard method in laboratories.

国家863计划项目（2013AA065802）;国家海洋局海洋可再生能源专项资金资助项目（GHME2011SW04）；广东省教育部产学研结合重点项目（2011A090200073）