If you are looking for optimized HPLC method to analyze 2-Hydroxyisocaproic Acid (HICA) check our HPLC Applications library

For optimal results in HPLC analysis, it is recommended to measure absorbance at a wavelength that matches the absorption maximum of the compound(s) being analyzed. The UV spectrum shown can assist in selecting an appropriate wavelength for your analysis. Please note that certain mobile phases and buffers may block wavelengths below 230 nm, rendering absorbance measurement at these wavelengths ineffective. If detection below 230 nm is required, it is recommended to use acetonitrile and water as low UV-transparent mobile phases, with phosphoric acid and its salts, sulfuric acid, and TFA as buffers.
For some compounds, the UV-Vis Spectrum is affected by the pH of the mobile phase. The spectra presented here are measured with an acidic mobile phase that has a pH of 3 or lower.

HPLC Method for β‑Hydroxy-β-methylbutyric Acid (HMB), 2-Hydroxyisocaproic Acid (HICA) on Primesep 100 by SIELC Technologies

 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.

 

Condition

Column	Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended
Mobile Phase	MeCN – 5%
Buffer	H2SO4 – 0.05 %
Flow Rate	1.0 ml/min
Detection	UV, 210 nm

 

Description

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 Method for β‑Hydroxy-β-methylbutyric Acid (HMB), D-Leucine, Leucine, 2-Hydroxyisocaproic Acid (HICA) on Primesep 100 by SIELC Technologies

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.