HPLC Method for Glycerol on Primesep 100 by SIELC Technologies
High Performance Liquid Chromatography (HPLC) Method for Analysis of Glycerol
Glycerol is an organic compound with the molecular formula C3H8O3. It is a humectant, solvent, and sweetener that used in various industries including skin care, hygiene products, pharmaceuticals, and food. It is said to improve lubrication and smoothness in products. Historically, before ethylene glycol took over, glycerol was used as a antifreeze. There is no Uv-Vis absorption found with standard Glycerol.
This derivatization was done using two reagents:
Reagent A (Periodate reagent): 6.5mg NaIO4 in 9mL of water, 1mL acetic acid, and 0.77g ammonium acetate
Reagent B (Acetylacetone reagent): 0.25mL of Acetylacetone in 24.75mL of isopropanol.
The sample was created by adding 0.2mL of Reagent A to 0.5mL hydrolysate and mixing for 5 minutes at room temperature. Afterwards, 0.5 mL of Reagent B was added to the solution and mixed for 20 minutes.
Glycerol 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 sulfuric acid buffer. Detection is performed using UV.
| Column | Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended |
| Mobile Phase | MeCN – 20% |
| Buffer | H2SO4 – 0.1% |
| Flow Rate | 1.0 ml/min |
| Detection | Vis 410 |
| Limit of Detection* | 2 ppm |
| Class of Compounds | Sugar |
| Analyzing Compounds | Glycerol |
Application Column
Primesep 100
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Column options: dual ended
