HPLC Method for Separation of Folic Acid, Methotrexate and Aminopterin on Primesep 100 Column
https://sielc.com/hplc-separation-of-aminopterin_folic-methotrexateChromatogram
Description
· Separation type: Liquid Chromatography Mixed-mode
Aminopterin and folic acid are chemically related compounds that play significant roles in biochemistry and medicine.
Methotrexate, Aminopterin, and Folic Acid are chemically related compounds that have important roles in biochemistry and medicine.
The antifolate drugs methotrexate and aminopterin are designed to interfere with the actions of folic acid, making them effective in slowing down the growth of rapidly dividing cells, such as cancer cells. However, this can also affect normal, healthy cells, leading to side effects.
Aminopterin, methotrexate and folic acid can be retained, separated, and analyzed using a reverse-phase Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended column. The mobile phase for this method consists of water, acetonitrile (MeCN), and ammonium formate, which serves as a buffer. This analytical method can be monitored using UV detection at 290 nm, an Evaporative Light Scattering Detector (ELSD), or any other evaporative detection method such as Charged Aerosol Detection (CAD) or Electrospray Ionization Mass Spectrometry (ESI-MS)
LOD was determined for this combination of instrument, method, and analyte, and it can vary from one laboratory to another even when the same general type of analysis is being performed.
Method Parameters
| Mobile Phase | Gradient MeCN -10-70%, 10 min |
|---|---|
| Buffer | Ammonium formate pH 3.0 – 50 mM |
| Flow Rate | 1.0 ml/min |
| Detection | UV 290 nm |
| Peak Retention Time | 5.08 min, 8.32 min |
| Sample concentration | 0.02 mg/ml |
| Injection volume | 5 µl |
| Sample diluent | H2O+NaOH |
| LOD | 0.04 ppm |
| Class of Compounds | Drug, Aminopterin, Antifolates, Acid |
| Analyzing Compounds | Methotrexate,Aminopterin,Folic Acid |