Comparative Study of the Separation and Quantification of Lipid Classes Separated on Chromarods‐A and Chromarods‐SIII by Thin Layer Chromatography with Iatroscan (Mark‐III) Flame Ionization Detection (TLC‐FID)

Abstract

Thin layer chromatography (TLC) on Chromarods‐A and –SIII were used for the separation of common lipid classes which were detected and quantified using Iatrosan Mark‐III with Flame Ionization Detection (FID). Two different lots from each type of Chromarods were used and half of each lot (5 rods) was considered as 1 analytical unit. Variability of the FID response for 11 different lipid classes (neutral and polar) among rods of each lot, among different analytical units as well as the variability between 2 different lots of the same type of Chromarods were compared statistically. Variability of the response for these lipid classes by an individual rod during repeated analysis was also investigated. Calibration curves of the FID response vs. sample loads were used to develop corresponding calibration models that can be used for quantification of lipid classes separated on these Chromarods.

There was no significant difference (p>0.05) in the FID response for all lipid classes separated on Chromarods‐A or ‐SIII. However, the FID responses for all lipid classes separated on Chromarods‐A were significantly different from those on Chromarods‐SIII (p<0.01). There was no significant difference in the FID response among analytical units of the same type of Chromarods. Curvilinear relationship of the FID response against the sample load was common to all lipid classes for both Chromarods‐A and –SIII. Several calibration equations were derived and Power Law models (y=ax^b) appear to be the most convenient and appropriate for rapid quantitative analysis of lipid classes although an addition of a quadratic and/or cubic term further improved the statistical validity of the model.

Keywords:

• Lipid classes
• Chromarods
• TLC‐FID
• Quantification
• Iatroscan‐Mark‐III