Animal cholesterol synthesis shows a marked diurnal variation, a phenomenon, at the moment, not known to occur in man. Since cholesterol precursors in serum reflect overall cholesterol synthesis in many conditions, a 24-hr profile of squalene and methyl sterols was studied in plasma lipoproteins in order to demonstrate whether these cholesterol precursors could exhibit a diurnal cycling in healthy human subjects. During the 24-hr period, lipoproteins of density < 1.006 g/ml transported 30-50% of the plasma squalene. Free methyl sterols were found mainly in low density lipoproteins (LDL) and esterified methyl sterols in LDL and high density lipoproteins (HDL). Postprandial hyperlipidemia at noon was associated with an inconsistent increase of the squalene and free methyl sterol concentrations in the lipoproteins of density < 1.006 g/ml. In terms of micro g per mg of cholesterol, the precursor contents were, however, low in each lipoprotein during the daytime. During the night and early morning, the values were several times higher. Thus the peak plasma squalene and methyl sterol contents occurred at midnight and 4 am. The highest variation was found for squalene in the density class < 1.006 g/ml and for lanosterol and diunsaturated dimethyl sterol in LDL and HDL. For different methyl sterols, the mean diurnal variation was 3.5- to 6.9-fold in LDL, 2.0- to 4.5-fold in HDL, and 2.6- to 3.6-fold in the density class < 1.006 g/ml. The respective values for squalene were 2.2, 1.4, and 2.9. Esterified methyl sterols varied slightly in the density class < 1.006 g/ml only, and the percentage esterification exhibited a diurnal fluctuation that was the reciprocal of that of free methyl sterol levels. The rapid and marked diurnal fluctuation of squalene and free methyl sterols in plasma lipoproteins suggests that these precursors are metabolized on and off lipoproteins. The variation is most likely caused by changes in cholesterol synthesis, inferring that circadian rhythm also regulates human cholesterol production.-Miettinen, T. A. Diurnal variation of cholesterol precursors squalene and methyl sterols in human plasma lipoproteins.