Analysis of High Molecular Weight Protein According to USP-NF GENERAL CHAPTERS Physicochemical Analytical Procedures for Insulins (KW-802.5)

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).

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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. 1.High molecular weight proteins
  2. 2.Insulin dimer
  3. 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 ℃

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Operation Manual / Certificate of Analysis

Operation Manuals and Certificate of Analysis / Inspection Certificate for the following products can be downloaded here.

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