Separation of Polar Solutes with HPLC – AppNote

Niacin, Riboflavin, Folic Acid, Pyridoxine, Metformin, Thiamine

The Cogent Diol Column is a good addition to the TYPE-C™ Silica line of HPLC stationary phases. Here, a variety of common polar analytes are well-retained and separated.

Peaks:
1. Ascorbic acid
2. Niacin
3. Riboflavin
4. Folic acid
5. Pyridoxine
6. Metformin
7. Thiamine

Method Conditions
Column: Cogent Diol™, 4μm, 100Å
Catalog No.: 40060-15P-3
Dimensions: 3.0 x 150mm
Mobile Phase:
A: DI Water / 0.1% Formic Acid (v/v)
B: Acetonitrile / 0.1% Formic Acid (v/v)
Gradient:

Time (minutes) %B
0 95
3 95
10 40
12 40
13 95

Post Time: 5 minutes
Flow rate: 0.7 mL/minute
Detection: UV @ 254 nm
Injection vol.: 1μL
Sample Preparation: Mixture of reference standards in diluent of 50 / 50 Solvent A / Solvent B.
t0: 0.7 minutes

Note: B and C Vitamins are hydrophilic and therefore may be difficult to retain in Reversed Phase methods. Metformin is a highly polar compound used for treatment of type 2 diabetes.

Attachment

No 291 Separation of Polar Test Solutes pdf  0.3 Mb  Download File