Comparative Study of Chemical Stability of a PK20 Opioid–Neurotensin Hybrid Peptide and Its Analogue [Ile9]PK20—The Effect of Isomerism of a Single Amino Acid
Abstract
:1. Introduction
2. Results
PK20 and [Ile9]PK20 Degradation Depending on Conditions
3. Discussion
4. Materials and Methods
4.1. Drugs and Chemicals
4.2. LC Apparatus and LC-MS Conditions
4.3. The Stock Solution of Investigated Compounds
4.4. Quantification of Peptides Using LC-MS
4.5. Method Validation
4.6. Degradation and Analysis Procedures of the Stressed Compounds
4.6.1. Acid and Base Hydrolysis
4.6.2. Thermal Degradation
4.6.3. UV Degradation
4.6.4. Oxidative Degradation
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Condition | Drug Concentration Injected (μg/mL) | Concentration Found (Mean ± SD, μg/mL) | Recovery (%) | RSD (%) * | |||
---|---|---|---|---|---|---|---|
PK20 | [Ile9]PK20 | PK20 | [Ile9]PK20 | PK20 | [Ile9]PK20 | ||
Acidic degradation (1 M HCl) | |||||||
37 °C (24 h) | 50 | 39.09 ± 6.33 | 44.39 ± 4.48 | 78.19 | 88.79 | 16.18 | 10.10 |
80 °C (12 h) | 50 | 27.22 ± 2.75 | 26.12 ± 1.52 | 54.43 | 52.25 | 10.09 | 5.83 |
Basic degradation (1 M NaOH) | |||||||
37 °C (24 h) | 50 | 15.16 ± 0.34 | 18.56 ± 0.59 | 30.32 | 37.13 | 2.27 | 3.19 |
80 °C (12 h) | 50 | 5.71 ± 1.67 | 5.61 ± 1.83 | 11.44 | 11.23 | 29.17 | 32.69 |
Oxidative degradation (30% H2O2) | |||||||
37 °C (24 h) | 50 | 30.52 ± 0.49 | 31.39 ± 0.27 | 61.04 | 62.79 | 1.60 | 0.87 |
80 °C (12 h) | 50 | 9.80 ± 0.2 | 8.24 ± 6.69 | 19.60 | 16.48 | 2.07 | 81.14 |
Thermal degradation | |||||||
Room temperature (24 h) | 50 | 51.36 ± 1.64 | 49.71 ± 2.24 | 102.72 | 99.41 | 3.19 | 4.50 |
37 °C (24 h) | 50 | 49.98 ± 0.35 | 44.16 ± 0.99 | 99.97 | 88.32 | 0.70 | 2.24 |
80 °C (12 h) | 50 | 48.67 ± 2.74 | 43.27 ± 1.18 | 97.34 | 86.54 | 5.63 | 2.73 |
−80 °C (24 h) | 50 | 51.90 ± 0.20 | 51.87 ± 0.40 | 103.80 | 103.74 | 0.38 | 0.77 |
Two freeze–thaw cycles (24 h (−80 °C), –2 h (22 °C), 24 h (−20 °C)) | 50 | 43.25 ± 9.42 | 49.04 ± 1.03 | 86.50 | 98.08 | 21.78 | 2.09 |
Photolytic degradation (365 nm UV light 7 h) | 50 | 18.72 ± 0.94 | 9.63 ± 0.11 | 37.44 | 19.25 | 5.04 | 1.14 |
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Żyżyńska-Granica, B.; Mollica, A.; Stefanucci, A.; Granica, S.; Kleczkowska, P. Comparative Study of Chemical Stability of a PK20 Opioid–Neurotensin Hybrid Peptide and Its Analogue [Ile9]PK20—The Effect of Isomerism of a Single Amino Acid. Int. J. Mol. Sci. 2022, 23, 10839. https://doi.org/10.3390/ijms231810839
Żyżyńska-Granica B, Mollica A, Stefanucci A, Granica S, Kleczkowska P. Comparative Study of Chemical Stability of a PK20 Opioid–Neurotensin Hybrid Peptide and Its Analogue [Ile9]PK20—The Effect of Isomerism of a Single Amino Acid. International Journal of Molecular Sciences. 2022; 23(18):10839. https://doi.org/10.3390/ijms231810839
Chicago/Turabian StyleŻyżyńska-Granica, Barbara, Adriano Mollica, Azzurra Stefanucci, Sebastian Granica, and Patrycja Kleczkowska. 2022. "Comparative Study of Chemical Stability of a PK20 Opioid–Neurotensin Hybrid Peptide and Its Analogue [Ile9]PK20—The Effect of Isomerism of a Single Amino Acid" International Journal of Molecular Sciences 23, no. 18: 10839. https://doi.org/10.3390/ijms231810839