Hradec J, Dufek P, Matucha M
Department of Cellular Biochemistry, Institute of Hematology and Blood Transfusion, Academy of Sciences of the Czech Republic, Prague.
J Chromatogr B Biomed Sci Appl. 1998 Sep 4;714(2):145-51. doi: 10.1016/s0378-4347(98)00189-3.
The multiplicity of phosphatidylcholines is caused by the presence of different pairs of fatty acids in their individual molecular species and at least 27 miscellaneous fatty acids were identified in phosphatidylcholines in the serum of healthy individuals by combined gas-liquid chromatography and mass spectrometry in our present experiments. A method is described for the separation and quantitation of molecular species of phosphatidylcholine in human serum. Total phosphatidylcholine is isolated from lipids extracted from the serum with chloroform-methanol (2:1) by reversed-phase liquid-liquid extraction and subjected to reversed-phase high-performance liquid chromatography with a discontinuous descending gradient of water. Separation is monitored by fluorometry (340/460 nm) and absorption at 205 nm, if required. Up to 25 different molecular species of phosphatidylcholine may be quantified with a satisfactory reproducibility (+/-5-8%). Data on the distribution of individual molecular species in phosphatidylcholine of 53 normal serums are presented. The method may be used for quantitation of these phospholipids also in other biological materials (cell lines, leukemic cells from patients), and on a micropreparative scale to isolate individual compounds. The speed of separation as well as a satisfactory reproducibility are its principal advantages.
磷脂酰胆碱的多样性是由其各个分子种类中存在不同的脂肪酸对所致，在我们目前的实验中，通过气相色谱 - 质谱联用技术在健康个体血清中的磷脂酰胆碱中鉴定出了至少27种不同的脂肪酸。本文描述了一种分离和定量人血清中磷脂酰胆碱分子种类的方法。总磷脂酰胆碱通过反相液 - 液萃取从用氯仿 - 甲醇（2:1）从血清中提取的脂质中分离出来，并采用水的不连续递减梯度进行反相高效液相色谱分析。如有需要，通过荧光法（340/460 nm）和205 nm处的吸光度监测分离情况。多达25种不同的磷脂酰胆碱分子种类可被定量，重现性良好（±5 - 8%）。给出了53份正常血清中磷脂酰胆碱各个分子种类的分布数据。该方法也可用于定量其他生物材料（细胞系、患者的白血病细胞）中的这些磷脂，并且可在微量制备规模上分离单个化合物。分离速度以及良好的重现性是其主要优点。
