HPLC Separation of β‑Hydroxy-β-methylbutyric Acid and 2-Hydroxyisocaproic Acid on Primesep 100 Colimn
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HPLC Separation of β‑Hydroxy-β-methylbutyric Acid and 2-Hydroxyisocaproic Acid_1182
Description
High Performance Liquid Chromatography (HPLC) Method for Analysis of β‑Hydroxy-β-methylbutyric Acid (HMB) , 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) , 2-Hydroxyisocaproic Acid (HICA) 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.
Method Parameters
| Mobile Phase | MeCN – 5% |
|---|---|
| Buffer | H2SO4 – 0.05 % |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 210 nm |
| Class of Compounds | Acid, Hydrophilic, Ionizable, Normal human metabolite, Dietary supplement, Essential amino acid, Branched-chain amino acid (BCAA) |
| Analyzing Compounds | β‑Hydroxy-β-methylbutyric Acid (HMB),2-Hydroxyisocaproic Acid (HICA) |
HPLC Column Used
Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended