微藻总脂含量测定方法概述

A Review of the Methods for Determining Total Lipid Content in Microalgae

概述微藻总脂含量测定方法的研究现状并比较各方法的优缺点:(1)有机溶剂提取法使用较普遍、易操作、成本低,缺点是所需样品量大、所得总脂含量较低、溶剂有毒易污染环境、操作耗时费力;(2)索氏提取法测定微藻总脂含量较准确,缺点是所需试剂和样品量大,提取时间长;(3)荧光染料测定法、傅里叶变换红外光谱法、磷酸香草醛法、铜试剂法、苏丹黑染色法以及核磁共振法所需样品量少、所用时间短,但测定的总脂含量均为相对值。其中,荧光染料测定法不需要提取油脂且灵敏度高,但其易受各种因子影响;傅里叶变换红外光谱法和核磁共振法对样品无损伤,无需或仅需很少的预处理,后者重复性很好;磷酸香草醛法较稳定;铜试剂法成本低,但对其报道极少;苏丹黑染色法操作简单,不需要对藻细胞破碎及化学抽提,但不适用于不均匀样品的测定;(4)超临界CO2提取法和离子液体提取法提取效率高,所得总脂含量较高,但测定成本较高。

The different methods for determining total lipid content in microalgae are reviewed in this paper,and the advantages and methodological limitations of those methods are compared： 1） Organic solvents extraction method is widely used because of easy operation and low cost.But it is inefficient and labor-consuming,requiring more samples,obtaining lower amount of extracted oil and using toxic solvents which could cause environment pollution.2） Soxhlet extraction method is more accurate for determining total lipid in microalgae.However,this method requires more reagents and samples,and long extraction time.3） Fluorescent dye method,FTIR spectroscopy method,Sulfo-Phospho-Vanillin（SPV） method,copper reagent method,Sudan black method and TD-NMR method all require only small volumes of sample,could be measured rapidly,but the total lipid content of all these methods are relative values.Among them,fluorescent dye method is sensitive and does not require lipid extraction,but vulnerable to be influenced by a variety of factors.FTIR spectroscopy method and TD-NMR method（which has good repeatability） are non-destructive,requiring little or no sample pretreatment.SPV method is stable.Copper reagent method is low cost,but there are very few reports about it until now.Sudan black method is simple,not requiring cell disruption and extraction,but not suitable for determination of uneven sample.4） Supercritical CO2 and ionic liquid methods are efficient in total lipid extraction,but the determination cost is high.