Stability Studies of Clonazepam 2.5 mg/mL Oral Solution and 1 mg/mL Parenteral Solution in Pre-Filled Polypropylene Syringes
Abstract
1. Introduction
2. Materials and Methods
2.1. Sample Preparation of Pre-Filled Syringes
2.2. Chemical Stability
2.3. Chromatographic Method
2.4. Physical Stability
3. Results
3.1. Validation of the Analytical Method
3.2. Stability Study
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
| D0 | Day zero of the test |
| D30 | Day thirty of the test |
| GABA | Gamma aminobutyric acid |
| HPLC | High-performance liquid chromatography |
| ICH | International Conference on Harmonisation |
| INSST | Spanish National Institute for Safety and Health at Work |
| LOD | Limit of detection |
| LOQ | Limit of quantification |
| NIOSH | National Institute for Occupational Safety and Health of the United States |
| PP | Polypropylene |
| R2 | Correlation coefficient |
| RSD% | Relative standard deviation |
| SD | Standard deviation |
| SEFH | Spanish Society of Hospital Pharmacy |
References
- Spanish Medicines and Medical Devices Agency Datasheet of Rivotril 1 mg/mL Concentrate and Solvent for Injectable Solution. Available online: https://cima.aemps.es/cima/dochtml/ft/52332/FT_52332.html (accessed on 12 August 2024).
- Spanish Medicines and Medical Devices Agency Datasheet of Rivotril 2.5 mg/mL Oral Drops Solution. Available online: https://cima.aemps.es/cima/dochtml/ft/52333/FT_52333.html#5-propiedades-farmacol-gicas (accessed on 12 August 2024).
- National Institute for Occupational Safety and Health Hazardous Drugs. Preventive Measures for Their Preparation and Administration. Available online: https://www.insst.es/documentacion/catalogo-de-publicaciones/medicamentos-peligrosos.-medidas-de-prevencion-para-su-preparacion-y-administracion (accessed on 12 August 2024).
- Connor, T.H.; MacKenzie, B.A.; DeBord, D.G.; Trout, D.B.; O’Callaghan, J.P.; Ovesen, J.L.; Whittaker, C.; Cincinnati, O.H.; National Institute for Occupational Safety and Health NIOSH [2020]. NIOSH List of Hazardous Drugs in Healthcare Settings 2020. Available online: https://www.cdc.gov/niosh/docket/review/docket233c/pdfs/DRAFT-NIOSH-Hazardous-Drugs-List-2020.pdf (accessed on 5 August 2024).
- Lal, R. Drugs in Pregnancy and Lactation: Improved Benefit-Risk Information 2015. Available online: https://www.fda.gov/files/drugs/published/%22Drugs-in-Pregnancy-and-Lactation--Improved-Benefit-Risk-Information%22-January-22--2015-Issue.pdf (accessed on 27 August 2024).
- Tuárez-Párraga, M.A.; Laz-Mero, M.; Córdova-Mosquera, A.; Panchana-Cedeño, R.; Gavilánes-López, P.; Solórzano Zambrano, L.; Tuárez-Párraga, M.A.; Laz-Mero, M.; Córdova-Mosquera, A.; Panchana-Cedeño, R.; et al. Oxygen Permeability in Food Packaging made of Polypropylene by Injection Moldin. Rev. Politécnica 2024, 53, 27–36. [Google Scholar] [CrossRef]
- Raimundo Piñero, A.; Selva Otaolarruvchi, J. Interacción de fármacos y mezclas parenterales con productos sanitarios. Panor. Actual Del Medicam. 2020, 11, 587–592. [Google Scholar]
- Laboratorios Grifols, S.A. Estabilidad y Compatibilidad de Medicamentos. Available online: https://gruposdetrabajo.sefh.es/gps/images/stories/descargas/Compatibilidad_medicamentos_diluyentes.pdf (accessed on 9 October 2024).
- European Directorate for the Quality of Medicines & HealthCare. European Pharmacopoeia, 11th ed.; European Directorate for the Quality of Medicines & HealthCare: Strasburgo, France, 2022. [Google Scholar]
- Abraham, J. Stability Testing of New Drug Sjustances and Products Q1A(R2). In International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; Tietje, C., Brouder, A., Eds.; Brill|Nijhoff: Leiden, The Netherlands, 2010; pp. 1041–1053. ISBN 978-90-04-18156-4. [Google Scholar]
- Troy, D.B.; Beringer, P. Remington: The Science and Practice of Pharmacy; Lippincott Williams & Wilkins: Philadelphia, PA, USA, 2006; ISBN 978-0-7817-4673-1. [Google Scholar]
- Spanish Agency of Medicines and Medical Devices Visible Particle Contamination. Spanish Royal Pharmacopoeia Online. Available online: https://extranet.boe.es/farmacopea/doc.php?id=20920 (accessed on 10 September 2024).
- Barrueco, N.; Escobar Rodríguez, I.; García Díaz, B.; Gil Alegre, M.E.; López Lunar, E.; Ventura Valares, M.G. The stability of medicines in the clinical practice. From safety to efficiency. Farm. Hosp. 2013, 37, 175–177. [Google Scholar] [CrossRef] [PubMed]
- United States Pharmacopeial Convention. United States Pharmacopeia and National Formulary (USP-NF), 2022nd ed.; US Pharmacopeia Convention, Inc.: Rockville, MD, USA, 2022. [Google Scholar]
- Guchelaar, H.J.; Hartog, M.E. De Stabiliteit van Clonazepaminjectievloeistof. Available online: https://www.stabilis.org/Bibliographie.php?IdBiblio=2217 (accessed on 11 August 2025).
- Castro, M.; Gascón, S.; Pujol, M.; Sans, J.; Vicente, L. Validation of Analytical Methods. Monograph. Commission on Good Manufacturing Practice and Quality Control Standards; Spanish Association of Industrial Pharmacists (AEFI): Barcelona, Spain, 1989. [Google Scholar]
- Allen, L.V.; Erickson, M.A. Stability of acetazolamide, allopurinol, azathioprine, clonazepam, and flucytosine in extemporaneously compounded oral liquids. Am. J. Health Syst. Pharm. 1996, 53, 1944–1949. [Google Scholar] [CrossRef] [PubMed]
- Polonini, H.C.; Loures, S.; Lima, L.C.; Ferreira, A.O.; Brandão, M.A.F. Stability of Atenolol, Clonazepam, Dexamethasone, Diclofenac Sodium, Diltiazem, Enalapril Maleate, Ketoprofen, Lamotrigine, Penicillamine-D, and Thiamine in SyrSpend SF PH4 Oral Suspensions. Int. J. Pharm. Compd. 2016, 20, 167–174. [Google Scholar] [PubMed]
- Abraham, J. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. In Handbook of Transnational Economic Governance Regimes; Brouder, A., Tietje, C., Eds.; Brill: Leiden, The Netherlands, 2009; pp. 1041–1054. ISBN 978-90-04-16330-0. [Google Scholar]
- Sagar Baliram, P.M. A Validated Stability–Indicating HPLC Method estimation of Clonazepam In the bulk drug and Pharmaceutical Dosage Form. Pharm. Anal. Acta 2015, 6, 2. [Google Scholar] [CrossRef]
- Hoffman, F. Statistical Validation: Quantitative determination (General Explanations); La Roche: Basel, Switzerland, 1987; pp. 1–9. [Google Scholar]
- Miller, J.; Miller, J.C. Statistics and Chemometrics for Analytical Chemistry; Pearson Education Limited: Harlow, UK, 2005; ISBN 84-205-3514-1. [Google Scholar]
- European Agency for Safety and Health at Work Guidance for the Safe Management of Hazardous Medicinal Products at Work | Safety and Health at Work EU-OSHA 2023. Available online: https://osha.europa.eu/en/publications/guidance-safe-management-hazardous-medicinal-products-work (accessed on 28 August 2024).
- Shaarma, S. Comprehensive stability evaluation and shelf -life determination of escitalopram and clonaepam fixed-dose combination tablets. IJHMP 2025, 6, 135–142. [Google Scholar] [CrossRef]
- Maślanka, A.; Krzek, J.; Stolarczyk, M.; Walczak, M.; Głogowska, A. Stability studies of clonazepam, diazepam, haloperidol, and doxepin with diverse polarities in an acidic environment. J. AOAC Int. 2011, 94, 1791–1799. [Google Scholar] [CrossRef] [PubMed]






| Theoretical Concentration (µg/mL) | 20 (80%) | 25 (100%) | 30 (120%) |
|---|---|---|---|
| Mean (µg/mL) | 20.04 | 24.99 | 30.04 |
| SD | 0.03 | 0.02 | 0.21 |
| RSD% | 0.15 | 0.09 | 0.68 |
| Accuracy% | 100.20 | 99.98 | 100.12 |
| Theoretical Concentration (µg/mL) | 20 (80%) | 25 (100%) | 30 (120%) |
|---|---|---|---|
| Mean (µg/mL) | 20.04 | 25.04 | 30.10 |
| SD | 0.07 | 0.06 | 0.21 |
| RSD% | 0.33 | 0.23 | 0.71 |
| Accuracy% | 100.21 | 100.15 | 100.32 |
| Room Temperature Not Protected from Light | Refrigeration Conditions Protected from Light | |||
|---|---|---|---|---|
| D0 | D30 | D0 | D30 | |
| Average Recovery % of concentration | 100 | 100.33 ± 0.01 | 100 | 97.82 ± 0.02 |
| pH | 4.63 ± 0.02 | 4.65 ± 0.06 | 4.65 ± 0.03 | 4.65 ± 0.05 |
| Color | Blue | Blue | Blue | Blue |
| Crystals ≥10 µm/mL | 0 | 0 | 0 | 0 |
| Room Temperature Not Protected from Light | Room Temperature Protected from Light | Refrigeration Conditions Protected from Light | ||
|---|---|---|---|---|
| D0 | D30 | D30 | D30 | |
| Average Recovery % of concentration | 100 | 100.87 ± 0.01 | 98.14 ± 0.02 | 98.02 ± 0.02 |
| pH | 4.15 ± 0.08 | 4.27 ± 0.15 | 4.30 ± 0.07 | 4.17 ± 0.04 |
| Color | Transparent | Transparent | Transparent | Transparent |
| Crystals ≥10 µm/mL | 0 | 0 | 0 | 0 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Ruiz Ramirez, J.C.; Talsi Hamdani, I.; Bermúdez Gazquez, L.; Viney, A.C.; Alonso Herreros, J.M. Stability Studies of Clonazepam 2.5 mg/mL Oral Solution and 1 mg/mL Parenteral Solution in Pre-Filled Polypropylene Syringes. Pharmaceutics 2025, 17, 1302. https://doi.org/10.3390/pharmaceutics17101302
Ruiz Ramirez JC, Talsi Hamdani I, Bermúdez Gazquez L, Viney AC, Alonso Herreros JM. Stability Studies of Clonazepam 2.5 mg/mL Oral Solution and 1 mg/mL Parenteral Solution in Pre-Filled Polypropylene Syringes. Pharmaceutics. 2025; 17(10):1302. https://doi.org/10.3390/pharmaceutics17101302
Chicago/Turabian StyleRuiz Ramirez, Juan Carlos, Icram Talsi Hamdani, Laura Bermúdez Gazquez, Alice Charlotte Viney, and José M. Alonso Herreros. 2025. "Stability Studies of Clonazepam 2.5 mg/mL Oral Solution and 1 mg/mL Parenteral Solution in Pre-Filled Polypropylene Syringes" Pharmaceutics 17, no. 10: 1302. https://doi.org/10.3390/pharmaceutics17101302
APA StyleRuiz Ramirez, J. C., Talsi Hamdani, I., Bermúdez Gazquez, L., Viney, A. C., & Alonso Herreros, J. M. (2025). Stability Studies of Clonazepam 2.5 mg/mL Oral Solution and 1 mg/mL Parenteral Solution in Pre-Filled Polypropylene Syringes. Pharmaceutics, 17(10), 1302. https://doi.org/10.3390/pharmaceutics17101302

