Application Notes

Fat Soluble Vitamins by HPLC/ELSD

Five fat-soluble vitamins were separated by reversed phase HPLC and detected by evaporative light scattering.

Fat-Soluble Vitamins, Alltech Application Note 0007E, June 23, 2000.

A mixture of five fat-soluble vitamins were separated isocratically in around 21 minutes by reversed phase HPLC on an Adsorbosphere C18, 5µm, 250 x 4.6mm column (Part No. 287062) using methanol/acetonitrile after first dissolving them in chloroform. Vitamins A, D2, D3, E and K, were detected by evaporative light scattering using an Alltech 500 ELSD (Evaporative Light Scattering Detector.)

In ELSD, the mobile phase is first evaporated. Solid particles remaining from the sample are then carried in the form of a mist into a cell where they are detected by a laser.

Key Analyte(s)

vitamin A

Vitamin A, an essential nutrient required for growth and vision is obtained commercially from fish oil. It is most often measured using spectrophotometry and HPLC.

vitamin D2

Vitamin D2 is an essential nutrient that maintains plasma calcium and phosphate levels. It is most often measured using colorimetry and HPLC.

vitamin D3

Vitamin D3 is an essential nutrient that facilitates caclium and phosphate absorption. It is most often measured using colorimetry and HPLC.

vitamin E

Vitamin E (alpha-tocopherol) is an antioxidant found in vegetable oils. It is most often measured using GC/FID and HPLC.

vitamin K

Vitamin K, synthetically derived from 2-methyl-1,4-naphthoquinone and phytol is used to treat blood coagulation disorders. It is most often measured using GC/FID and HPLC.

Equipment

Alltech Adsorbosphere Columns

Alltech Adsorbosphere® Columns are designed using high temperature bonding for exceptional surface coverage and stability for common applications. They are available in pore sizes from 60-90Å and come in four varieties.

Technique

HPLC

High Performance Liquid Chromatography (HPLC)is a technique for separating components in a sample on the basis of interactions between the component, a liquid mobile phase and a solid stationary phase.

Source

Alltech Associates, Inc.

Alltech Associates, Inc., one of the leading suppliers of HPLC columns, manufactures and distributes HPLC, GC and IC columns, accessories and consumables and maintains a large searchable database of application notes and chromatograms on its website.