一种用于测定组织中磷酸多萜醇的高效液相色谱法。

A high-performance liquid chromatographic method for the determination of dolichyl phosphates in tissues.

作者信息

Yamada K, Abe S, Suzuki T, Katayama K, Sato T

出版信息

Anal Biochem. 1986 Aug 1;156(2):380-5. doi: 10.1016/0003-2697(86)90268-x.

Abstract

A sensitive high-performance liquid chromatographic method has been developed for the determination of dolichyl phosphate homologs in tissues. Dolichyl phosphates were methylated and derivatized with 3-(9-anthryl)-diazo-2-propene to yield methyl 3-(9-anthryl)-2-propenyl dolichyl phosphates. As an internal standard, 2,2-didecaprenylethyl phosphate was used. The derivatives were separated on a reversed-phase column using isocratic elution, and detected fluorometrically. The method with fluorescence detection was sufficiently sensitive to measure the concentration of dolichyl phosphate homologs in tissues.

摘要

已开发出一种灵敏的高效液相色谱法用于测定组织中的磷酸多萜醇同系物。磷酸多萜醇经甲基化并用3-(9-蒽基)-重氮-2-丙烯进行衍生化，生成3-(9-蒽基)-2-丙烯基磷酸多萜醇甲酯。使用2,2-二癸戊烯基乙基磷酸酯作为内标。衍生物在反相柱上采用等度洗脱进行分离，并用荧光法检测。这种荧光检测方法灵敏度足够高，能够测定组织中磷酸多萜醇同系物的浓度。