title: LC-MS/MS analysis of canine lipoproteins fractionated using the ultracentrifugation-precipitation method.
abstract: Due to the lack of a gold standard method in canine lipoprotein analysis, it is unclear whether canine high-density lipoprotein (HDL) and low-density lipoprotein (LDL) can be accurately evaluated by the lipoprotein analysis methods used for dogs. This study investigated whether the ultracentrifugation-precipitation (U-P) method was suitable as a gold standard method for analyzing canine lipoprotein. First, the U-P method was compared with a gel permeation high-performance liquid chromatography system (GP-HPLC). The concentrations of canine HDL cholesterol (HDL-C) and LDL cholesterol (LDL-C) determined by the U-P method correlated closely with those determined by GP-HPLC. However, the canine HDL-C concentration determined by the U-P method was lower than that determined by GP-HPLC, and the canine LDL-C concentration determined by the U-P method was higher than that determined by GP-HPLC. This study showed that some canine HDL could be precipitated with heparin manganese chloride solution. Second, the HDL and LDL fractions separated by the U-P method were analyzed by LC-MS/MS. The HDL fraction was found to contain only apolipoprotein A-I, which is an apolipoprotein of HDL, whereas the LDL fraction contained both apolipoprotein A-I and apolipoprotein B-100, which is an apolipoprotein of LDL. This data showed that a certain lipoprotein that includes apolipoprotein A-I might precipitate with canine LDL when using heparin manganese chloride solution. These results indicated that the U-P method is not currently a gold standard method for analyzing canine lipoproteins.
authors.affiliation: Department of Dermatology, Hokkaido University Graduate School of Medicine, Kita15 Nishi7, Kita-ku, Sapporo, Hokkaido 060-8638, Japan.
authors.forename: Asuka
authors.initials: A
authors.lastname: Suto
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yamasaki
authors.forename: Yukari
authors.initials: Y
authors.lastname: Takasaki
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Fujita
authors.forename: Riichiro
authors.initials: R
authors.lastname: Abe
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimizu
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ohta
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Takiguchi
mesh_terms: D000818:Animals; D011232:Chemical Precipitation; D002853:Chromatography, Liquid / Q000662:veterinary; D004285:Dogs / Q000097:blood*; D005260:Female; D008075:Lipoproteins, HDL / Q000097:blood*; D008077:Lipoproteins, LDL / Q000097:blood*; D008297:Male; D053719:Tandem Mass Spectrometry / Q000662:veterinary; D014461:Ultracentrifugation / Q000379:methods / Q000662:veterinary*
chemical_list: D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL