Quantitation of Total Cholesterol and HDL-Cholesterol by the HPLC Method: Comparison with the Automated Enzymatic Method and the Precipitation Method

Sasamoto Keiko, Okazaki Mitsuyo, Muramatsu Toshio, Yanagisawa Tsutomu, Ando Yuriko, Kamei Sachiko, Hosaki Seijin

doi:10.14921/jscc1971b.25.1_28

Total cholesterol (TC) and HDL-cholesterol (C) of human sera (n=134) from subjects with serum triglyceride (TG) levels from 40mg/dl to 5378mg/dl were measured by an improved HPLC system using a gel filtration column (TSK gel Lipopropak, 7.5mm×600mm/TOSOH Co. Tokyo) and an eluent (TSKeluent LP-1). The values were compared with TC measured by the automated enzymatic method (HITACHI 7070) and HDL measured by the precipitation method. By the HPLC method, TC and HDL-C levels as well as the HDL sizes could be simultaneously and accurately obtained from the HPLC patterns monitored by cholesterol for all subjects including severe hypertriglyceridemia (n=7), in which HDL could not be separated by the precipitation method. Although the TC levels obtained by the HPLC method were consistent with those obtained by the automated enzymatic method, HDL-C levels were slightly higher than those obtained by the precipitation method. The HDL of all the samples with discrepant values between the two methods were revealed to be large in size and rich in cholesterol. Therefore, HDL subclasses with a larger size of particle and higher amount of cholesterol than normal HDL were suggested to be precipitated selectively by the precipitation method. A negative correlation was obtained between the HDL-C level by the two methods and serum TG level, but correlations were improved by excluding the samples with discrepant HDL-C levels between the two methods.

Quantitation of Total Cholesterol and HDL-Cholesterol by the HPLC Method: Comparison with the Automated Enzymatic Method and the Precipitation Method

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