HPLC Method for Separation of Methotrexate, Aminopterin, Folic Acid on Primesep 100 by SIELC Technologies
High Performance Liquid Chromatography (HPLC) Method for Analysis of Methotrexate, Aminopterin, Folic Acid.
Methotrexate is a chemotherapy agent and immune system suppressant with the chemical formula C20H22N8O5. It is an antimetabolite and antifolate drug, meaning it inhibits the metabolism of folic acid. It does this by inhibiting the enzyme dihydrofolate reductase (DHFR), preventing the formation of active folate and thus interfering with DNA and RNA synthesis. It is used to treat certain types of cancer and autoimmune diseases like rheumatoid arthritis and psoriasis. You can find detailed UV spectra of Methotrexate and information about its various lambda maxima by visiting the following link.
Aminopterin is also an antifolate with the chemical formula C19H20N8O5. It works in a similar way to methotrexate, by inhibiting the enzyme DHFR. Aminopterin was one of the first antifolates to be used in cancer chemotherapy. However, it has largely been replaced by methotrexate, which is less toxic and more effective. You can find detailed UV spectra of Aminopterin and information about its various lambda maxima by visiting the following link.
Folic Acid, also known as vitamin B9, folic acid is crucial for many functions in the body, including DNA synthesis and repair, cell division, and growth. As an essential vitamin, Folic acid must be obtained through diet or supplementation, as humans cannot synthesize it. Vitamin B-9 is a crucial prenatal vitamin. that also helps the body use iron properly. It has the chemical formula C19H19N7O6. You can find detailed UV spectra of Folic Acid and information about its various lambda maxima by visiting the following link.
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.
Methotrexate, Aminopterin, Folic Acid can be retained, separated, and analyzed using a reverse-phase Primesep 100 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.
Condition
| Column | Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended |
| 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 |
Description
| Class of Compounds | Drug, Aminopterin, Antifolates, Acid |
| Analyzing Compounds | Methotrexate, Aminopterin, Folic Acid |
Application Column
Primesep 100
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Column options: dual ended
Folic Acid
Methotrexate
