HPLC Separation of β‑Hydroxy-β-methylbutyric Acid and 2-Hydroxyisocaproic Acid on Primesep 100 Colimn

HPLC-Applikation • Datum: 2019/09/20
HPLC Separation of β‑Hydroxy-β-methylbutyric Acid and 2-Hydroxyisocaproic Acid on Primesep 100 Colimn Chromatogramm

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.

Methodenparameter

ColumnPrimesep 100, 4.6 x 150 mm, 5 µm, 100 Å, dual ended
Mobile PhaseMeCN – 5%
BufferH2SO4 – 0.05%
Flow Rate1.0 mL/min
DetectionUV, 210 nm