Suto Asuka, Yamasaki Masahiro, Takasaki Yukari, Fujita Yasuyuki, Abe Riichiro, Shimizu Hiroshi, Ohta Hiroshi, Takiguchi Mitsuyoshi
Department of Dermatology, Hokkaido University Graduate School of Medicine, Kita15 Nishi7, Kita-ku, Sapporo, Hokkaido 060-8638, Japan.
J Vet Med Sci. 2013 Nov;75(11):1471-7. doi: 10.1292/jvms.13-0098. Epub 2013 Jul 16.
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.
由于犬脂蛋白分析中缺乏金标准方法，目前尚不清楚用于犬的脂蛋白分析方法能否准确评估犬高密度脂蛋白（HDL）和低密度脂蛋白（LDL）。本研究调查了超速离心沉淀法（U-P法）是否适合作为分析犬脂蛋白的金标准方法。首先，将U-P法与凝胶渗透高效液相色谱系统（GP-HPLC）进行比较。U-P法测定的犬HDL胆固醇（HDL-C）和LDL胆固醇（LDL-C）浓度与GP-HPLC测定的浓度密切相关。然而，U-P法测定的犬HDL-C浓度低于GP-HPLC测定的浓度，U-P法测定的犬LDL-C浓度高于GP-HPLC测定的浓度。本研究表明，部分犬HDL可被肝素氯化锰溶液沉淀。其次，通过液相色谱-串联质谱法（LC-MS/MS）分析U-P法分离出的HDL和LDL组分。发现HDL组分仅含有HDL的载脂蛋白A-I，而LDL组分同时含有LDL的载脂蛋白A-I和载脂蛋白B-100。这些数据表明，使用肝素氯化锰溶液时，某些包含载脂蛋白A-I的脂蛋白可能会与犬LDL一起沉淀。这些结果表明，U-P法目前并非分析犬脂蛋白的金标准方法。
