HPLC Method for Analysis of Biuret on Primesep S Column
Separation type: Liquid Chromatography Mixed-mode SIELC Technologies
Biuret, with the chemical formula C2H5N3O2, is a white crystalline compound formed by the condensation of two urea molecules with the release of ammonia. Its structure consists of two amide groups (-CONH) linked by a nitrogen atom. Biuret is widely used in biochemistry for protein assays, specifically in the biuret test, which detects peptide bonds. Due to its polar functional groups, the compound is highly soluble in water.
Biuret can be retained, separated and analyzed using a Primesep S mixed-mode stationary phase column. The analysis employs an isocratic method with a simple mobile phase comprising water, acetonitrile (MeCN), and phosphoric as a buffer. This method allows for detection using UV 200 nm.
Methodenparameter
| Column | Primesep S, 4.6 x 150 mm, 5 µm, 100 Å, dual ended |
|---|---|
| Mobile Phase | MeCN – 90% |
| Buffer | H3PO4 – 0.1% |
| Flow Rate | 1.0 mL/min |
| Detection | UV 200 nm |