Analysis of Methylene Blue Purity in Reversed-Phase Cation-Exchange and Reversed-Phase Cation-Exclusion Modes
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.
Methodenparameter
| Column | Primesep C, 4.6×150 mm, 5 µm, 100 Å |
|---|---|
| Mobile Phase | MeCN – 35% |
| Buffer | AmFm pH 3.5 – 35 mM |
| Flow Rate | 1.0 mL/min |
| Detection | UV, 300 nm |