The use of the Iatroscan TH-10 analyzer to quantify total lipids in a variety of sample types and lipid classes in human gallbladder bile.
Two methods for the measurement of total lipid weight in biological and geological samples and the major lipid classes in human gallbladder bile using the Iatroscan TH-10 analyzer are described. Total lipid determination involves the application of small (5 microliter) volumes to Chromarods, focusing of the sample into one band by partial development in chloroform-methanol (1:1), and quantification by flame ionization detection (FID). The response variation between different sample types did not affect the linearity of response, allowing a reproducibility of +/- 10% of the mean or better for samples ranging from 0.5 to 32 micrograms. Total lipid determinations in 10 samples could be performed in 30 min. The three major components of human gallbladder bile (cholesterol, phospholipids and bile acids) also were quantified with the Iatroscan. Samples focused on Chromarods were separated using a double development scheme in two solvent systems. All three components exhibited a linear response over the range of 0.25 to 8 micrograms. The repeated scanning of rods required at concentrations greater than 3 micrograms did not affect linearity of response. Samples from 10 patients could be processed in less than one hr. Several techniques are discussed to increase reproducibility when performing quantitative lipid analysis with the Iatroscan.
D001646:Bile / Q000032:analysis*; D001647:Bile Acids and Salts / Q000032:analysis; D002784:Cholesterol / Q000032:analysis; D002855:Chromatography, Thin Layer / Q000379:methods; D005410:Flame Ionization / Q000379:methods; D005704:Gallbladder / Q000032:analysis; D006801:Humans; D008055:Lipids / Q000032:analysis*
D001647:Bile Acids and Salts; D008055:Lipids; D002784:Cholesterol