HPLC Separation of β‑Hydroxy-β-methylbutyric Acid, 2-Hydroxyisocaproic Acid and Leucine on Primesep 100 Column
High Performance Liquid Chromatography (HPLC) Method for Analysis of β‑Hydroxy-β-methylbutyric Acid (HMB) , D-Leucine , Leucine , 2-Hydroxyisocaproic Acid (HICA)
Both β-hydroxy-β-methylbutyric acid (HMB) and 2-hydroxyisocaproic acid (HICA) are metabolites of the essential dietary branched chain amino acid leucine.
β‑Hydroxy-β-methylbutyric Acid (HMB) , D-Leucine , Leucine , 2-Hydroxyisocaproic Acid (HICA) can be retained and analyzed using the Primesep 100 stationary phase column. The analysis utilizes a gradient method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a sulfuric acid buffer. Detection is performed using UV.
Methodenparameter
| Column | Primesep 100, 4.6 x 150 mm, 5 µm, 100 Å, dual ended |
|---|---|
| Mobile Phase | MeCN – 5%, 3 min hold, Gradient MeCN5-30% 12 min |
| Buffer | H2SO4 – 0.05% 3 min hold, GradientH2SO4 0.05-3% 12 min |
| Flow Rate | 1.0 mL/min |
| Detection | UV, 210 nm |