Quantitative detection method of triglycerides in serum lipoproteins and serum-free glycerol by high-performance liquid chromatography.
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.
Okazaki
M
M
Laboratory of Chemistry, College of Liberal Arts and Sciences, Tokyo Medical and Dental University, Ichikawa, Japan.
Komoriya
N
N
Tomoike
H
H
Inoue
N
N
Usui
S
S
Itoh
S
S
Hosaki
S
S
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005990:Glycerol / Q000097:blood* / Q000378:metabolism; D005991:Glycerol Kinase; D005993:Glycerolphosphate Dehydrogenase; D006801:Humans; D006949:Hyperlipidemias / Q000097:blood; D008071:Lipoprotein Lipase; D008074:Lipoproteins / Q000097:blood / Q000737:chemistry*; D008297:Male; D011485:Protein Binding; D011817:Rabbits; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors; D014280:Triglycerides / Q000097:blood*
D001798:Blood Proteins; D008074:Lipoproteins; D014280:Triglycerides; D005993:Glycerolphosphate Dehydrogenase; C098831:glycerol-3-phosphate oxidase; D005991:Glycerol Kinase; D008071:Lipoprotein Lipase; D005990:Glycerol