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Compound Evidence InstituteEvidence synthesis · established 2023Graded assessments of compounds, trials, methods and supply
Document set current to 30 July 2026
Methodological standard · Stability

CEI-MS-21 — Freeze–thaw and in-use stability assessment of reconstituted peptide solutions

Freeze–thaw and in-use stability assessment of reconstituted peptide solutions

Document identifier
CEI-MS-21
Series
Methodological standard
Version
1.0
Published
16 Jun 2026
Last reviewed
16 Jun 2026
Next review
16 Jun 2029
Identifier
10.71829/cei.meth.18
Certainty
High
Cycle
2026 Q2
Category
Stability

§1Scope

§1.1Scope

Applies to the assessment of any in-use period claimed for a reconstituted peptide solution, and to the practice of freezing reconstituted solutions for later use. The Institute publishes no in-use claim for reconstituted research material and this standard states what would be required to support one.

§2Principle

Freezing a peptide solution concentrates solutes in the shrinking liquid fraction, raising local ionic strength and driving pH shifts as buffer components crystallise differentially. Phosphate buffers are particularly prone to a large pH drop on freezing. Ice–liquid interfaces denature and aggregate proteins and amphipathic peptides. These effects are not detected by a reverse-phase purity determination and require an aggregate determination alongside.[1,2]

§3Apparatus and reagents

§3.1Apparatus

  • Controlled-rate freezer or validated −20 °C and −80 °C storage
  • Refrigerated storage at 2–8 °C with continuous monitoring
  • pH meter suitable for small volumes
  • HPLC and size-exclusion systems as CEI-MS-01 and CEI-MS-11

§3.2Reagents and reference materials

  • Diluent as used in practice, including bacteriostatic water where that is the practice being assessed
  • Reference standard

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 Q1A(R2) Stability Testing of New Drug Substances and Products. ICH Harmonised Tripartite Guideline 2003;Step 4 version. identifier not held by the Institute
  2. Manning MC, Chou DK, Murphy BM, Payne RW, Katayama DS. Stability of protein pharmaceuticals: an update. Pharmaceutical Research 2010;27(4):544–575. doi:10.1007/s11095-009-0045-6 · PMID 20143256

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