Yan Kun-Ping, Hao Jing, Dan Ning, Chen Chao
College of Life Science, Northwest University, Xi'an, PR China.
Artif Cells Blood Substit Immobil Biotechnol. 2008;36(1):19-33. doi: 10.1080/10731190701857751.
This study introduced a liquid-liquid extraction method designed for complete recovery of phospholipids from protein-rich samples, such as hemoglobin (Hb) solutions. In order to minimize protein denaturation and maximize lipid extraction from protein-rich samples, isopropanol was used as the major organic extraction solvent. In a wide range of the volume ratio of isopropanol to Hb solution at low temperatures, such extraction system resulted in limited protein precipitation and did not cause heme pigment contamination. The efficiency of phospholipid extraction from Hb solutions was pH-dependent, with the lowest extraction yield around neural pH and the highest extraction yield around pH 5.0. At pH 5.0, salt with concentrations up to 400 mmol/L in the samples did not increase extraction efficiency. Compared to other available methods, this method is simpler, needs significantly less organic solvent and, most importantly, consumed much less Hb, which is very expensive to make for the purpose of production of therapeutic products, hemoglobin-based oxygen carriers (HBOCs). This method is suitable not only for Hb solutions but also for other protein-rich biological samples, such as erythrocytes, plasma and liver.
本研究介绍了一种液-液萃取方法，该方法旨在从富含蛋白质的样品（如血红蛋白（Hb）溶液）中完全回收磷脂。为了使蛋白质变性最小化并使从富含蛋白质的样品中提取的脂质最大化，异丙醇被用作主要的有机萃取溶剂。在低温下，在异丙醇与Hb溶液的广泛体积比范围内，这种萃取系统导致有限的蛋白质沉淀，并且不会引起血红素色素污染。从Hb溶液中提取磷脂的效率取决于pH值，在接近中性pH时提取产率最低，在pH 5.0左右时提取产率最高。在pH 5.0时，样品中浓度高达400 mmol/L的盐不会提高提取效率。与其他现有方法相比，该方法更简单，所需有机溶剂显著更少，最重要的是，消耗的Hb少得多，而Hb用于生产治疗产品、基于血红蛋白的氧载体（HBOCs）时制备成本非常高。该方法不仅适用于Hb溶液，也适用于其他富含蛋白质的生物样品，如红细胞、血浆和肝脏。
