Different animal models have been used to study the pathomechanisms involved in dyslipidemia and evaluate the effect of drugs on serum lipid.However, most of these models differ from human beings in the responses to high-fat and high-cholesterol diet, this may be caused by variations in lipoprotein distribution among different species.Lipoprotein profiles of different dyslipidemia animal model have not been explored systematically due to the lack of an appropriate method.Traditional ultra-centrifugal lipoprotein separation technique and the commonly used liquid chromatography methods both have their disadvantages in lipoprotein separation, lipoprotein and lipid component quantification.In this study, we combined small volume needed size-exclusion fast protein liquid chromatography (FPLC) and fluorescence lipids measurement, accomplished the determination of serum lipoprotein profiles in C57 BL/6 Mice, ApoE-/-Mice, SD Rats, Golden Hamster, Guinea Pigs, Rabbits, Miniature Pigs and Human Beings.Our results demonstrated that there were considerable variations in the FPLC spectrum, lipoprotein distribution and lipoprotein subclasses/lipoprotein ratio among these different individuals.In dyslipidemia patients, decreased high density lipoprotein (HDL)/lipoprotein ratio and increased low density lipoprotein (LDL)/lipoprotein ratio were observed, but it was found that the changes of HDL/lipoprotein and LDL/lipoprotein ratios in SD Rats after high-fat and high-cholesterol diet were totally different with Human Beings, meanwhile, its HDL/Lipoprotein ratio was increased, and LDL/Lipoprotein ratio was decreased.Whereas Golden Hamster, Miniature Pigs and Human Beings have much in common in lipoprotein distribution and response to high-fat and high-cholesterol diet.This was the first time that two different size-exclusion columns were combined and used to separate small volume individual serum lipoprotein.Since fluorescence detection methods were used to measure lipid components in the eluted lipoprotein fractions, it became possible that determination of lipoprotein profiles and gaining information of lipids in different lipoproteins could be accomplished simultaneously.It was likely that variations in diet, expression of lipoprotein and activity of lipids metabolism associated enzymes in different species may result in the variations of lipoprotein profiles.It inspires us to explore the exactly mechanism that produces the variations of lipoprotein profiles in different species, the association among the variations and the different response, the attack time and severity of atherosclerosis in different species after high-fat and high-cholesterol diet.