Analysis of Methylene Blue Purity in Reversed-Phase Cation-Exchange and Reversed-Phase Cation-Exclusion Modes
https://sielc.com/Application-Analysis-of-Methylene-Blue-Purity-in-Reversed-Phase-Cation-Exchange-and-Reversed-Phase-Cation-Exclusion-ModesChromatogram
Description
Methylene blue is a heterocyclic dye with a wide range of use in biology and chemistry. Purity of dyes rarely exceed 70% and separation of methylene blue and related impurities is required. Two alternative methods for purity analysis were developed on Primesep C and Primesep D mixed-mode HPLC columns. Primesep C is a reversed-phase cation-exchange (anion-exclusion column) column and Primesep D is reversed-phase anion-exchange (cation-exclusion) column. Various buffers can be used for the method. Depending on the buffer, method can be compatible with UV, LC/MS and prep chromatography. Method can be used for prep separation of methylene blue and related impurities.
Method Parameters
| Mobile Phase | MeCN – 35% |
|---|---|
| Buffer | AmFm pH 3.5 – 35 mM |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 300 nm |
| Class of Compounds | Duy, Hydrophobic, Ionizable |
| Analyzing Compounds | Methylene Blue |
HPLC Column Used
Primesep C, 4.6×150 mm, 5 µm, 100A