Rapid micromethod for flourometric determination of neutral lipids on thin-layer chromatograms.
A rapid fluorometric procedure for in situ evaluation of neutral lipids on thin-layer chromatograms is described. The lipids are rendered fluorescent by treatment with perchloric acid, and the intensity of the emitted light is recorded by flying spot scanning. Detection limits (2.8 x SD) are 0.10, 0.11, and 0.17 nmol for triglycerides, cholesterol, and cholesterol esters, respectively. Range of linearity is 0--3.4 nmol for triglycerides, 0--2.0 nmol for cholesterol, and 0--4.0 nmol for cholesterol esters. When standard in triplicate and 2 samples of each unknown are applied to the chromatogram, standard error of the mean is 2.7% for triglycerides, 1.7% for cholesterol, and 1.0% for cholesterol esters. The sensitivity of the method allows simultaneous determination of the three lipid classes in 5 microliter lipid extract representing 0.238 microliter human plasma.
D000818:Animals; D002784:Cholesterol / Q000097:blood; D002788:Cholesterol Esters / Q000097:blood; D002855:Chromatography, Thin Layer; D005470:Fluorometry / Q000379:methods*; D008055:Lipids / Q000097:blood*; D010743:Phospholipids / Q000097:blood; D051381:Rats; D014280:Triglycerides / Q000097:blood
D002788:Cholesterol Esters; D008055:Lipids; D010743:Phospholipids; D014280:Triglycerides; D002784:Cholesterol