High Performance Liquid Chromatography (HPLC) Method for Analysis of Crystal Violet on Primesep 100 by SIELC Technologies.
Separation type: Liquid Chromatography Mixed-mode
Crystal Violet, also known as Methyl Violet 10B, is a synthetic dye belonging to the triarylmethane dye family. It has the molecular formula C25H30ClN3. This dye is primarily known for its use in microbiology as a stain, but it has other applications as well.
- Gram Staining: One of the most well-known uses of Crystal Violet is in Gram staining, a method used to differentiate bacterial species into two large groups (Gram-positive and Gram-negative) based on the chemical and physical properties of their cell walls. In this process, a crystal violet solution is applied to a smear of bacterial cells, which are then treated with iodine, followed by an alcohol wash and counterstain with safranin. Gram-positive bacteria retain the crystal violet stain, while Gram-negative bacteria do not.
- Antiseptic: Crystal Violet has antiseptic properties and has been used as a topical antiseptic to treat minor burns, infections, and fungal conditions. However, due to concerns over toxicity, its use in this application has declined.
- Cell Biology and Histology: In cell biology and histology, Crystal Violet is used to stain cell nuclei, allowing for visualization under a microscope. It is also used in the assessment of cell viability and proliferation in various assays, such as the crystal violet cell cytotoxicity assay.
- Indicator in Chemistry: In analytical chemistry, Crystal Violet is used as an indicator in colorimetric titrations due to its ability to change color at different pH levels.
- Textile and Paper Industry: Crystal Violet is also used as a dye in the textile and paper industry.
Crystal Violet can be retained, and analyzed on a Primesep 100 mixed-mode stationary phase column using an isocratic analytical method with a simple mobile phase of water, Acetonitrile (MeCN), and a sulfuric acid (H2SO4) buffer. This analysis method can be detected in the UV-Vis regime at 540, 590, and 200 nm.
High Performance Liquid Chromatography (HPLC) Method for Analyses of Crystal Violet
Condition
| Column | Primesep 100, 4.6 x 150 mm, 5 µm, 100 A |
| Mobile Phase | MeCN/H2O – 80/20% |
| Buffer | H3PO4 – 0.2% |
| Flow Rate | 1.0 ml/min |
| Detection | Vis, 590 nm |
| Peak Retention Time | 9.37 min |
| Sample concentration | 0.0006 mg/ml |
| Injection volume | 5 µl |
| LOD | 10 ppb |
Description
| Class of Compounds | Dyes |
| Analyzing Compounds | Crystal Violet |
Application Column
Primesep 100
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Application Analytes:
Crystal VioletApplication Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.
