Analysis of Insulin Human According to JP Method (KW-802.5)

According to JP (Japanese Pharmacopoeia) method, high-molecular proteins in insulin human (genetical recombination) should be analyzed using a column packed with hydrophilic silica gel. It is necessary for the system suitability to satisfy the ratio, H₁/H₂ ≥ 2.0_. Where H₁ represents the peak height of the dimer and H₂ represents the height of the bottom between the peaks of the dimer and the monomer. It was confirmed that H₁/H₂ was more than 30 when they were analyzed using PROTEIN KW-802.5.

Sample : 100μL
4.0mg/mL of human Insulin containing dimer (in 0.01N HCl aq.)
1. High molecular weight proteins
2. Insulin trimer
3. Insulin dimer
4. Insulin monomer

Column       : Shodex PROTEIN KW-802.5 (8.0mmI.D. x 300mm)
Eluent       : 0.1wt% L-Arginine aq./CH3CN/CH3COOH=13/4/3
Flow rate    : 0.5mL/min
Detector     : UV(276nm)
Column temp. : 25°C