Separation of allo-D-isoleucine and L- Isoleucine on Primesep 200

HPLC Method for L-Isoleucine, allo-D-isoleucine on Primesep 200 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of L-Isoleucine, allo-D-isoleucine 

Allo-D-isoleucine is a non-proteinogenic amino acid with C6H13NO molecular formula. It is most commonly used in protein synthesis, neuroprotection, and immune modulation. It is also crucial in synthesis of peptides and can support muscle growth and recovery.

L- Isoleucine is a branched-chain amino acid with C6H13NO2 molecular formula. During chemical reactions, it serves as a nitrogen donor. It is also involved in glucose consumption, fatty acid metabolism, and immune functions. It can be found in a variety of animal and plant products, including meat, eggs, dairy, soybeans, lentils, peas, nuts, and seeds. Healthcare providers may encourage athletes and people with muscle atrophy to take it as a supplement.  Excessive consumption of it may lead to side effects including nausea, vomiting, and diarrhea.

L-Isoleucine, allo-D-isoleucine can be retained and analyzed using the Primesep 200 stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a trifluoroacetic acid buffer. Detection is performed using CAD.

ColumnPrimesep 200, 4.6×150 mm, 5 µm, 100A
Mobile PhaseMeCN/H2O
BufferTFA, FA
Flow Rate1.0 ml/min
DetectionCAD (Corona) MS- compatible mobile phase
Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid
Analyzing CompoundsAllo-Isoleucine, Isoleucine, Leucine

Application Column

Primesep 200

Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Column options: dual ended

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Application Analytes:
L-Isoleucine
allo-D-isoleucine

Application Detection:
CAD Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.