Horne T, Holt-Larkin S
Hemosol Inc., Etobicoke, Ont., Canada.
J Chromatogr B Biomed Sci Appl. 1997 Aug 1;695(2):259-67. doi: 10.1016/s0378-4347(97)00204-1.
Styrene-divinylbenzene Empore disks were investigated for the extraction of phospholipids from red blood cells or aqueous solutions of hemoglobin as a means to reduce the time and solvent use required in sample preparation. Red blood cells are the source for hemoglobin used in the preparation of a hemoglobin-based oxygen carrier which is being developed to replace blood in transfusion therapy. Phospholipids are a major component of the membrane of red blood cells, and are toxic when administered directly into the vasculature. Sensitive analytical methods are required to detect phospholipids to ensure that concentrations in purified hemoglobin are well below toxic levels. This requires isolation from large volumes of purified hemoglobin solutions. The method described utilizes Empore disks to extract phospholipids from 30 ml of stroma free Hb preparations. Phosphatidylethanolamine, phosphatidylinositol, phosphatidylcholine and sphingomyelin were recovered with an average of 92% yield. The recovery of phosphatidylserine was 65%. The use of solvent and time required for sample preparation were reduced by an average of 80% relative to liquid-liquid extraction. The capacity of the 47-mm disk for the total of five phospholipids exceeds 0.3 mg. The method has been used for quantitation of phospholipids in red blood cells and stroma free hemoglobin solutions.
研究了苯乙烯-二乙烯基苯Empore盘从红细胞或血红蛋白水溶液中提取磷脂的方法，以此作为减少样品制备所需时间和溶剂用量的手段。红细胞是制备基于血红蛋白的氧载体所用血红蛋白的来源，该氧载体正被开发用于替代输血治疗中的血液。磷脂是红细胞膜的主要成分，直接注入血管时具有毒性。需要灵敏的分析方法来检测磷脂，以确保纯化血红蛋白中的浓度远低于毒性水平。这需要从大量纯化血红蛋白溶液中分离出来。所述方法利用Empore盘从30毫升无基质血红蛋白制剂中提取磷脂。磷脂酰乙醇胺、磷脂酰肌醇、磷脂酰胆碱和鞘磷脂的回收率平均为92%。磷脂酰丝氨酸的回收率为65%。相对于液-液萃取，样品制备所需的溶剂用量和时间平均减少了80%。47毫米盘对五种磷脂的总容量超过0.3毫克。该方法已用于定量红细胞和无基质血红蛋白溶液中的磷脂。
