CEI-MS-11 — procedure

§4Procedure

The procedure below is written in the second person, which is the only place in the Institute's document set where that register is used. A procedure is an instruction and is written as one.

1. Equilibrate the column isocratically for not fewer than 10 column volumes and confirm baseline stability.
2. Calibrate with molecular-mass standards and record the retention-versus-mass relationship.
3. Prepare the sample in mobile phase without agitation. Filter only through a low-binding 0.22 µm membrane, and record that filtration itself can remove aggregates and therefore understate them.
4. Inject without delay. Non-covalent aggregate populations change over hours in solution.
5. Integrate the monomer peak and all species eluting earlier. Report high-molecular-weight species as a percentage of total area.
6. Where an absolute molar mass is required, acquire with light scattering and report the determined molar mass of each population rather than an apparent mass from calibration.
7. Report monomer content, high-molecular-weight species content, the mobile phase used, and whether the sample was filtered.

References cited on this page

References are numbered in order of first citation in this document. Each superscript in the text links to its entry below.

1. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products. ICH Harmonised Tripartite Guideline 1999;Step 4 version. identifier not held by the Institute

Identifiers are reproduced only where the Institute holds them. Where a digital object identifier or PubMed identifier is not shown, the Institute has recorded the journal and year and has not constructed an identifier.