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. 2022 Oct;12(5):774-782.
doi: 10.1016/j.jpha.2022.03.003. Epub 2022 Mar 24.

Effects of polyol excipient stability during storage and use on the quality of biopharmaceutical formulations

Min-Fei Sun  1   2 , Jia-Ning Liao  1 , Zhen-Yi Jing  1 , Han Gao  1   2 , Bin-Bin Shen  1   2 , You-Fu Xu  3 , Wei-Jie Fang  1   2
Affiliations

Effects of polyol excipient stability during storage and use on the quality of biopharmaceutical formulations

Min-Fei Sun et al. J Pharm Anal. 2022 Oct.

Abstract

Biopharmaceuticals are formulated using a variety of excipients to maintain their storage stability. However, some excipients are prone to degradation during repeated use and/or improper storage, and the impurities generated by their degradation are easily overlooked by end users and are usually not strictly monitored, affecting the stability of biopharmaceuticals. In this study, we evaluated the degradation profile of polyol excipient glycerol during repeated use and improper storage and identified an unprecedented cyclic ketal impurity using gas chromatography with mass spectrometry (GC-MS). The other polyol excipient, mannitol, was much more stable than glycerol. The effects of degraded glycerol and mannitol on the stability of the model biopharmaceutical pentapeptide, thymopentin, were also evaluated. The thymopentin content was only 66.4% in the thymopentin formulations with degraded glycerol, compared to 95.8% in other formulations after the stress test. Most glycerol impurities (i.e., aldehydes and ketones) reacted with thymopentin, affecting the stability of thymopentin formulations. In conclusion, this work suggests that more attention should be paid to the quality changes of excipients during repeated use and storage. Additional testing of excipient stability under real or accelerated conditions by manufacturers would help avoid unexpected and painful results.

Keywords: Excipient stability; GC-MS; Glycerol; LC-MS/MS; Mannitol; Thymopentin.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Image 1
Graphical abstract
Fig. 1
Fig. 1
Structures of polyol pharmaceutical excipients: (A) glycerol, and (B) mannitol.
Fig. 2
Fig. 2
Spectrophotometry with phenol reagent.
Fig. 3
Fig. 3
(A) Mass spectra of impurity 4 eluting at 7.4 min in gas chromatography with mass spectrometry (GC-MS). (B) The structure of impurity 4 and its mass spectrum fragments.
Fig. 4
Fig. 4
(A) The contents of aldehydes at 0 and 8 weeks of each formulation group. * P < 0.0001. (B) Degradation of thymopentin in different formulations at weeks 0–8 by reverse phase high performance liquid chromatography (RP-HPLC) (red); changes in aldehyde content in TP5-G1 formulations at weeks 0–8 (blue). (C) HPLC chromatogram of thymopentin formulations at the 8th week. (TP5-M0 refers to thymopentin formulations with undegraded mannitol; TP5-M1 refers to thymopentin formulations with degraded mannitol; TP5-G0 refers to thymopentin formulations with undegraded glycerol; TP5-G1 refers to thymopentin formulations with degraded glycerol; TP5-0H refers to untreated thymopentin formulations; mean ± SD, n = 3).
Fig. 5
Fig. 5
(A) MS/MS analysis of thymopentin as a representative. (B) MS/MS spectrum of thymopentin.
Fig. 6
Fig. 6
Changes in peak area of products 8a and 13 in weeks 0–8 of thymopentin formulations with degraded glycerol (mean ± SD, n = 3).
Fig. 7
Fig. 7
Conversion mechanism of products 8a and 13.
Fig. 8
Fig. 8
Effect of polyol excipient stability during storage and use on the quality of the thymopentin formulations. GC-MS: gas chromatography with mass spectrometry; LC-MS/MS: liquid chromatography coupled with tandem mass spectrometry.
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