Day R S, de Salamanca R E, Eales L
Clin Chim Acta. 1978 Oct 2;89(1):25-33. doi: 10.1016/0009-8981(78)90358-3.
This paper reports a new method for the quantitative determination of erythrocyte and plasma porphyrins by fluoroscanning of the methyl esters separated by thin-layer chromatography (TLC). After esterification overnight in the dark the polycarboxylated porphyrin methyl esters were extracted into chloroform and aliquots applied to the TLC plates within the range of quantities shown in a preliminary study to be directly proportional to fluorescence intensity. The accuracy and reliability of the technique was tested by comparison of RBC protoporphyrin values obtained by TLC with an established quantitative porphyrin solvent extraction method and with a rapid method presently in common use. Good correlation was demonstrated between the solvent extraction and the TLC techniques. The high sensitivity and adaptability of the TLC technique and its ability to clearly separate all the porphyrins present in the samples are discussed, along with the fluorescence mechanisms involved and the effects of instrumentation.
本文报道了一种通过对经薄层色谱（TLC）分离的甲酯进行荧光扫描来定量测定红细胞和血浆卟啉的新方法。在黑暗中过夜酯化后，将多羧化卟啉甲酯萃取到氯仿中，并将等分试样点样到TLC板上，其数量范围在初步研究中显示与荧光强度成正比。通过将TLC法获得的红细胞原卟啉值与既定的定量卟啉溶剂萃取法以及目前常用的快速方法进行比较，对该技术的准确性和可靠性进行了测试。结果表明溶剂萃取法与TLC技术之间具有良好的相关性。文中讨论了TLC技术的高灵敏度和适应性，以及它清晰分离样品中所有卟啉的能力，同时还讨论了相关的荧光机制和仪器的影响。
