High Performance Liquid Chromatography (HPLC) Method for Analysis of Eteplirsen on Primesep 100 by SIELC Technologies
Separation type: Liquid Chromatography Mixed-mode
Eteplirsen (brand name Exondys 51) is an antisense oligonucleotide used for the treatment of some types of Duchenne muscular dystrophy (DMD), a rare genetic disorder characterized by progressive muscle degeneration and weakness. Eteplirsen specifically targets patients who have a confirmed mutation of the DMD gene that is amenable to exon 51 skipping.
Mechanism of Action: Eteplirsen works by binding to exon 51 of dystrophin pre-mRNA. This results in the exclusion of exon 51 during mRNA processing, producing a truncated but still functional form of dystrophin protein.
Approval: Eteplirsen was granted accelerated approval by the U.S. Food and Drug Administration (FDA) in 2016. This approval was based on an increase in dystrophin protein in skeletal muscle observed in some patients treated with Eteplirsen. The FDA has required the manufacturer to conduct a clinical trial to confirm the drug’s clinical benefit.
Administration: Eteplirsen is administered as an intravenous infusion.
Chemical Structure: Eteplirsen is an oligonucleotide, a short sequence of nucleic acids. Its full chemical structure is detailed and specific, given its nature as an oligonucleotide.
Safety and Efficacy: Common side effects include balance disorder and vomiting. The clinical benefit of Eteplirsen, including improved motor function, has not been definitively established.
Eteplirsen can be retained, and analyzed on a Primesep B mixed-mode stationary phase column using an isocratic analytical method with a simple mobile phase of water, Acetonitrile (MeCN), and a sulfuric acid as a buffer. This analysis method can be detected using UV at 260 nm.
High Performance Liquid Chromatography (HPLC) Method for Analyses of Eteplirsen
Condition
| Column | Primesep B, 4.6 x 150 mm, 5 µm, 100 A |
| Mobile Phase | Gradient MeCN – 10-60%, 10 min |
| Buffer | H2SO4 – 0.2% |
| Flow Rate | 1.0 ml/min |
| Detection | UV 260 nm |
| Peak Retention Time | 6.26 min |
| LOD * | 94 ppb |
*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.
Description
| Class of Compounds | Oligonucleotide |
| Analyzing Compounds | Eteplirsen |
Application Column
Primesep B
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
