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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