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Excipient effect on phenol-induced precipitation of human growth hormone and bovine serum albumin

Hjalte, Johanna LU ; Börjesdotter, Anna Maria LU ; Diehl, Carl ; Ulvenlund, Stefan LU ; Wahlgren, Marie LU orcid and Sjögren, Helen (2025) In International Journal of Pharmaceutics 676.
Abstract

The aim of this study was to investigate the impact of phenol on the precipitation of bovine serum albumin (BSA) and human growth hormone (hGH), in the presence of other excipients frequently used in biological drugs for parenteral delivery. The focus of the study lieson incompatibilities observed in multidose formulations containing non-ionic surfactants and preservatives. Previous research has shown that above a critical concentration, phenol reduces the cloud point of polysorbate surfactants to room temperature or lower. Here, it is demonstrated that for BSA-polysorbate solutions, phenol-induced incompatibility is primarily controlled by this depression of the surfactant cloud point, resulting in turbidity and/or precipitation.... (More)

The aim of this study was to investigate the impact of phenol on the precipitation of bovine serum albumin (BSA) and human growth hormone (hGH), in the presence of other excipients frequently used in biological drugs for parenteral delivery. The focus of the study lieson incompatibilities observed in multidose formulations containing non-ionic surfactants and preservatives. Previous research has shown that above a critical concentration, phenol reduces the cloud point of polysorbate surfactants to room temperature or lower. Here, it is demonstrated that for BSA-polysorbate solutions, phenol-induced incompatibility is primarily controlled by this depression of the surfactant cloud point, resulting in turbidity and/or precipitation. However, for formulations with human growth hormone (hGH) in isotonic salt solutions, the precipitation mechanism is instead driven by protein-phenol interactions. The precipitation is affected by the concentration of sodium chloride and at low salt concentrations the incompatibility is again controlled by depression of the surfactant cloud point. The concentration of salt needed for protein induced precipitation seems to follow the Hofmeister series, with sodium chloride and sodium sulphate inducing precipitation at a lower salt concentration than sodium nitrate. Notably, non-ionic tonicity agents, such as glucose and mannitol, which are known to impact the surfactant cloud point depression of phenol, do not induce precipitation of hGH in the presence of phenol. In the system containing polysorbate, phenol and hGH, salt-triggered protein precipitation occurs at slightly higher sodium chloride concentrations than in solutions without polysorbate. This indicates a stabilizing effect of polysorbate on hGH below the cloud point. However, the stabilising effect is surfactant dependent, and in the presence of dodecyl maltoside, hGH precipitation occurs at much lower sodium chloride concentrations than for solutions with polysorbates. This illustrates the complexity of the interplay of excipients with each other and with the active ingredient (the protein) in the development of multidose pharmaceutics.

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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Clouding, Growth hormone, Incompatibility, Phenol, Precipitation, Preservative, Serum albumin
in
International Journal of Pharmaceutics
volume
676
article number
125624
publisher
Elsevier
external identifiers
  • pmid:40268209
  • scopus:105003114504
ISSN
0378-5173
DOI
10.1016/j.ijpharm.2025.125624
language
English
LU publication?
yes
id
84a26328-b9f1-4ed1-a71e-8a27e67cfa3a
date added to LUP
2025-08-04 11:49:05
date last changed
2025-08-05 03:08:28
@article{84a26328-b9f1-4ed1-a71e-8a27e67cfa3a,
  abstract     = {{<p>The aim of this study was to investigate the impact of phenol on the precipitation of bovine serum albumin (BSA) and human growth hormone (hGH), in the presence of other excipients frequently used in biological drugs for parenteral delivery. The focus of the study lieson incompatibilities observed in multidose formulations containing non-ionic surfactants and preservatives. Previous research has shown that above a critical concentration, phenol reduces the cloud point of polysorbate surfactants to room temperature or lower. Here, it is demonstrated that for BSA-polysorbate solutions, phenol-induced incompatibility is primarily controlled by this depression of the surfactant cloud point, resulting in turbidity and/or precipitation. However, for formulations with human growth hormone (hGH) in isotonic salt solutions, the precipitation mechanism is instead driven by protein-phenol interactions. The precipitation is affected by the concentration of sodium chloride and at low salt concentrations the incompatibility is again controlled by depression of the surfactant cloud point. The concentration of salt needed for protein induced precipitation seems to follow the Hofmeister series, with sodium chloride and sodium sulphate inducing precipitation at a lower salt concentration than sodium nitrate. Notably, non-ionic tonicity agents, such as glucose and mannitol, which are known to impact the surfactant cloud point depression of phenol, do not induce precipitation of hGH in the presence of phenol. In the system containing polysorbate, phenol and hGH, salt-triggered protein precipitation occurs at slightly higher sodium chloride concentrations than in solutions without polysorbate. This indicates a stabilizing effect of polysorbate on hGH below the cloud point. However, the stabilising effect is surfactant dependent, and in the presence of dodecyl maltoside, hGH precipitation occurs at much lower sodium chloride concentrations than for solutions with polysorbates. This illustrates the complexity of the interplay of excipients with each other and with the active ingredient (the protein) in the development of multidose pharmaceutics.</p>}},
  author       = {{Hjalte, Johanna and Börjesdotter, Anna Maria and Diehl, Carl and Ulvenlund, Stefan and Wahlgren, Marie and Sjögren, Helen}},
  issn         = {{0378-5173}},
  keywords     = {{Clouding; Growth hormone; Incompatibility; Phenol; Precipitation; Preservative; Serum albumin}},
  language     = {{eng}},
  publisher    = {{Elsevier}},
  series       = {{International Journal of Pharmaceutics}},
  title        = {{Excipient effect on phenol-induced precipitation of human growth hormone and bovine serum albumin}},
  url          = {{http://dx.doi.org/10.1016/j.ijpharm.2025.125624}},
  doi          = {{10.1016/j.ijpharm.2025.125624}},
  volume       = {{676}},
  year         = {{2025}},
}