Kolarovic L, Fournier N C
Anal Biochem. 1986 Jul;156(1):244-50. doi: 10.1016/0003-2697(86)90179-x.
Four classical methods, as well as a method presented in this paper, were compared as to their efficiency in extracting phospholipids from animal tissue. After the extractions, total lipids were separated quantitatively by DEAE-Sephadex chromatography into their acidic and nonacidic fractions. The two fractions were then further analyzed by gradient saturation high-performance thin-layer chromatography (HPTLC) combined with scanning photodensitometry after coloration with copper acetate. Of the five methods compared, the present and Christiansen's methods based upon single-phase solvent systems proved to be more efficient than biphasic extraction procedures. The undesirable discriminatory effect exhibited by biphasic solvent systems toward acidic phospholipids which were partly retained in the aqueous phase was confirmed by statistical evaluation of the HPTLC results. Total chromogenic response of acidic phospholipids extracted using biphasic solvent systems was shown to be lower by 10-35% in comparison to the single-phase method of Christiansen. The suitability of the present method for studies involving phospholipid synthesis was confirmed by monitoring the elimination of water-soluble compounds from the single-phase extracts using a classical phospholipid precursor, 2-[3H]glycerol-3-phosphate. The labeled compound was eliminated (99.3-100%) from the single-phase postcentrifugation supernatant, followed by DEAE-Sephadex chromatography.
对四种经典方法以及本文提出的一种方法在从动物组织中提取磷脂的效率方面进行了比较。提取后，通过DEAE-葡聚糖凝胶色谱法将总脂质定量分离为酸性和非酸性组分。然后，在用醋酸铜染色后，通过梯度饱和高效薄层色谱法（HPTLC）结合扫描光密度法对这两个组分进行进一步分析。在所比较的五种方法中，基于单相溶剂体系的本文方法和克里斯蒂安森方法被证明比双相萃取方法更有效。通过对HPTLC结果的统计评估证实了双相溶剂体系对部分保留在水相中的酸性磷脂表现出的不良歧视效应。与克里斯蒂安森的单相方法相比，使用双相溶剂体系提取的酸性磷脂的总显色响应低10 - 35%。通过使用经典磷脂前体2-[³H]甘油-3-磷酸监测单相提取物中水溶性化合物的去除情况，证实了本文方法适用于涉及磷脂合成的研究。标记化合物在单相离心后上清液中被去除（99.3 - 100%），随后进行DEAE-葡聚糖凝胶色谱法。
