TY - JOUR
T1 - Quantitative detection method of triglycerides in serum lipoproteins and serum-free glycerol by high-performance liquid chromatography
AU - Okazaki, Mitsuyo
AU - Komoriya, Norikazu
AU - Tomoike, Hitonobu
AU - Inoue, Noriko
AU - Usui, Shinichi
AU - Itoh, Sekiko
AU - Hosaki, Seijin
PY - 1998/5/29
Y1 - 1998/5/29
N2 - We have developed a simple and reliable method for quantitative detection of triglycerides (TG) in serum lipoproteins and serum-free glycerol (FG) by high-performance liquid chromatography (HPLC). After separation of serum constituents using a new gel-permeation column (TSK gel Lipopropak XL, Tosoh) and a new eluent (TSK eluent LP-2, Tosoh), TG and FG were detected by on-line reaction using a modified reagent which contained glycerol kinase, glycerol-3-phosphate oxidase and lipoprotein lipase. HPLC patterns showed five peaks corresponding to chylomicrons, very-low-density, low-density, high-density lipoproteins and FG. Absolute concentrations of TG in each lipoprotein fraction and serum FG were calculated from the corresponding peak areas using standard FG as a calibrator. Due to its very high sensitivity of peak detection, this method has become desirable for the analyses of lipoproteins of very low concentrations such as in cell culture systems. This technique will contribute to a better understanding of lipoprotein TG and serum FG distribution in human and nonhuman subjects.
AB - We have developed a simple and reliable method for quantitative detection of triglycerides (TG) in serum lipoproteins and serum-free glycerol (FG) by high-performance liquid chromatography (HPLC). After separation of serum constituents using a new gel-permeation column (TSK gel Lipopropak XL, Tosoh) and a new eluent (TSK eluent LP-2, Tosoh), TG and FG were detected by on-line reaction using a modified reagent which contained glycerol kinase, glycerol-3-phosphate oxidase and lipoprotein lipase. HPLC patterns showed five peaks corresponding to chylomicrons, very-low-density, low-density, high-density lipoproteins and FG. Absolute concentrations of TG in each lipoprotein fraction and serum FG were calculated from the corresponding peak areas using standard FG as a calibrator. Due to its very high sensitivity of peak detection, this method has become desirable for the analyses of lipoproteins of very low concentrations such as in cell culture systems. This technique will contribute to a better understanding of lipoprotein TG and serum FG distribution in human and nonhuman subjects.
KW - Free glycerol
KW - Lipoproteins
KW - Triglycerides
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U2 - 10.1016/S0378-4347(98)00064-4
DO - 10.1016/S0378-4347(98)00064-4
M3 - Article
C2 - 9657214
AN - SCOPUS:0032577545
SN - 1572-6495
VL - 709
SP - 179
EP - 187
JO - Journal of Chromatography B: Biomedical Applications
JF - Journal of Chromatography B: Biomedical Applications
IS - 2
ER -