We prepared several serum batches with various cholesterol concentrations, to be used as calibrators and controls in a proficiency testing program of an organization in The Netherlands that is in charge of the standardization of cholesterol determinations for epidemiological purposes. The sera were of human origin, to avoid abnormal matrix effects. To decrease the cholesterol content in some samples, we adsorbed them onto colloidal silicic acid. To increase it, we added lipoproteins that had been precipitated from human serum with heparin and calcium ions. The precipitation method we used allowed us to dilute the serum as little as possible and to keep the final concentrations of calcium and heparin as low as possible, while maintaining a high cholesterol content. By mixing these sera having high and low cholesterol concentrations, we could prepare batches with any desired concentration. The stabilities of these sera were excellent. We used the sera to calibrate enzymic determinations of cholesterol.