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HPLC Application Sheet
Nucleoside Analogs: Anticancer Prodrugs

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Nucleoside analogue anticancer “pro” drugs are difficult to retain on conventional HPLC columns because of their highly polar natures. With other HPLC Columns, ion pair reagents normally are used in order to achieve sufficient retention. These additives interfere with MS detection and are not preferred. With the Cogent UDC-Cholesterol column, you can have sufficient retention with a MS compatible mobile phase.

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Method Conditions



Column Cogent UDC Cholesterol™, 4mm, 100A
Catalog No. 69069-7.5P
Dimensions 4.6 x 75mm
Mobile Phase 60:40 acetonitrile/DI water + 0.5% formic acid
Flow rate 1.0 mL/minute
Injection Volume  5 µL
Samples Proprietary Drugs
N1 - N3 nucleoside analogues
P1 - P3 other more hydrophobic proprietary compounds with polar functional groups
1 mg/L of each dissolved in the mobile phase
Detection UV 270nm - Diode Array


Discussion


Cholesterol HPLC Columns are able to retain polar and nonpolar compounds in a single isocratic run. Adjustment of the aqueous to organic ratio can be used to make either the polar or more hydrophobic compounds elute first. In the example presented the more hydrophobic compounds are retained longer but retention and separation of the polar constituents is still achieved.

If greater retention of the polar compounds is desired then a higher percentage of acetonitrile is required.






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