USP-NF GENERAL CHAPTERS lists <121.1> Physicochemical Analytical Procedures for Insulins. The target analytes included in this method are insulin analogues, animal-derived insulin, and human insulin. The analysis of high molecular weight proteins should be carried out with a column filled with L20 packing material and meets following requirements.
System suitability requirements
Retention times:
Polymeric insulin complexes: 13 – 17 min
Covalent insulin dimer: about 17.5 min
Insulin monomer: 18 – 22 min
Peak-to-valley ratio*: ≥ 2.0
*The ratio of the height of the covalent insulin dimer peak to the height of the valley between the covalent insulin dimer peak and the insulin monomer peak.
Below lists the monographs using this method (at 2023 June).
| Insulin | Insulin Injection | Insulin Human | Insulin Human Injection |
| Insulin Aspart | Insulin Aspart Injection | Insulin Glargine | Insulin Glargine Injection |
| Insulin Lispro | Insulin Lispro Injection | Insulin Zinc Suspension | Extended Insulin Zinc Suspension |
| Prompt Insulin Zinc Suspension | Isophane Insulin Suspension | Isophane Insulin Human Suspension | Human Insulin Isophane Suspension and Human Insulin Injection |
The PROTEIN KW-802.5 confirmed the requirements were met for the analysis of insulin beef.
Sample: 100 µL
4.0 mg/mL of Insulin (beef) containing dimer (in 0.01 N HCl aq.)
- 1.High molecular weight proteins
- 2.Insulin dimer
- 3.Insulin monomer
- Column：Shodex PROTEIN KW-802.5 (8.0 mm I.D. x 300 mm)
- Eluent：0.1 wt% L-Arginine aq./CH3CN/CH3COOH=13/4/3
- Flow rate：0.5 mL/min
- Detector：UV (276 nm)
- Column temp.：25 ℃
