HPLC Method for Analysis of Thymol Blue on Primesep 100 Column
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Description
· High Performance Liquid Chromatography (HPLC) Method for Analysis of Thymol blue
Thymol Blue is a sulfonephthalein dye with C 27 H 30 O 5 S molecular formula. It is typically used as a pH indicator. Despite it’s name, it is only blue at higher pH levels, turning from blue to yellow when going basic to neutral and from yellow to red when changing from relatively acidic to very acidic. Due to the split in transition stages, it us considered a diprotic acid.
Thymol blue can be retained and analyzed using the Primesep 100 stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a formic acid buffer. Detection is performed using UV.
Method Parameters
| Mobile Phase | MeCN/H2O – 25/75% |
|---|---|
| Buffer | Formic Acid – 0.3 % |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 270 nm |
| Class of Compounds | Dye, Acid, Hydrophobic |
| Analyzing Compounds | Thymol blue |
HPLC Column Used
Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended