Density gradient ultracentrifugation in the swing out rotor has been successfully applied to fractionation of the major lipoprotein species and their subspecies; however, it requires a minimum of 24–36 h to complete the separation of the major lipoprotein fractions in plasma. This chapter describes 13 separate vertical spin procedures for the preparative separation of a number of different plasma lipoproteins and a method for the quantitative analysis of lipoprotein cholesterol from plasma separated by single vertical spin (SVS) density gradient ultracentrifugation. The vertical rotor works on the principle that compression of the gradient geometry by the vertical rotor when at speed shortens spin time over more conventional ultracentrifugation techniques without loss of resolution. The single vertical spin procedure has a number of advantages over the more conventional methods including decreased spin time, decreased lipoprotein degradation, and good resolution. But there are certain disadvantages of this method also, such as wall adherence of VLDL and albumin, resolution, and plasma volume limit per rotor. However, the chapter addresses solutions to the problems of single vertical spin ultracentrifugation such as angled-head rotors. © 1986, Elsevier Inc. All rights reserved.