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Unique Surface Treatment of Novel TSK-GEL ODS Columns Provide Separation of Aqueous and Fat Soluble Vitamins

ODS-100V than on the ODS-100Z column. Also, separation of nicotinamide and nicotinic acid was enhanced on the more polar column.


Analysis of Lipid Soluble Vitamins
Analysis of vitamin D2 (ergocalciferol) and D3 (cholecalciferol) is critical because they differ only in one methylene group and one double bond. As shown in Figure 2, separation was achieved under isocratic conditions in 100% acetonitrile at a flow rate of 1.0mL/min. The influence of temperature on resolution was evaluated at 25C and 40C.


For this separation TSKgel ODS-100Z, which has higher % carbon than TSKgel ODS-100V, provides better resolution. Ergocalciferol and cholecalciferol elute from both columns with near baseline separation. On the TSKgel ODS-100V column, hydrophobic retention is not high enough for a baseline separation of both vitamin D compounds. On both columns, separation is enhanced at lower temperature (25C).


Simultaneous Separation of Water and Lipid Soluble Vitamins
Water and lipid soluble vitamins were separated in one single run on TSKgel ODS-100V and TSKgel ODS-100Z as demonstrated in Figure 3 .


To separate the mixture of six water-soluble vitamins, caffeine, and four fat-soluble vitamins, required a gradient from water containing 0.1% TFA (eluent A) to acetonitrile containing 0.1% TFA (eluent B). Flow rate was 1mL/min, at a temperature of 40C. To elute the polar vitamins, the content of acetonitrile in the eluent was gradually increased up to 40% within 20 minutes, then, within 2 minutes, up to 100%, enabling elution of the less polar vitamins within another 27 minutes.

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