Application Notes
Water Soluble Vitamins by HPLC/ELSD
Four water-soluble vitamins were separated by reversed phase HPLC and detected by evaporative light scattering.
Water-Soluble Vitamins, Alltech Application Note 0006E, June 5, 2000.
A mixture of four water-soluble vitamins was separated in around 17 minutes by reversed phase HPLC on an Alltima C18, 5µm, 250 x 2.1mm column (Part No. 88371) using gradient elution with ammonium acetate in water and acetonitrile. The vitamins, ascorbic acid, niacin, thiamine and riboflavin 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)
ascorbic acid (Vitamin C)Ascorbic acid (Vitamin C) is an essential nutrient found in fruit juices and used as a vitamin supplement in food. It is most often measured using colorimetry and HPLC.
niacinNiacin (nicotinic acid)is an essential nutrient found in meats and cereals and used as a dietary supplement in foods. It is most often measured colorimetrically and by HPLC.
riboflavinRiboflavin (vitamin B2)is a vitamin found naturally in foods and added to enriched flours. It is most often measured using HPLC.
thiamine (Vitamin B1)Thiamin (Vitamin B1), a coenzyme essential for carbohydrate metabolism, is found in all plants and also produced synthetically. It is most often measured usng fluoresence and HPLC techniques.
Equipment
Alltech Alltima Reversed-Phase ColumnsAlltech Alltima Reversed Phase columns are a line of HPLC columns in the Alltech HP family. The Alltech Alltima family are high quality general purpose columns. They are made with spherical silica, polymerically bonded and double endcapped for long life
Technique
HPLCHigh 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.
