Stability-Indicating UPLC Method Development and Validation for Sulfamethoxazole and Trimethoprim Injection with Comprehensive Forced Degradation Profiling
Abstract
Objective: To develop and validate a stability-indicating UPLC method for the simultaneous estimation of Sulfamethoxazole and Trimethoprim in injectable dosage form, including comprehensive forced degradation profiling as per ICH guidelines.
Design: Experimental study involving method development, validation, and forced degradation in accordance with ICH Q2(R1) and Q1A(R2) protocols.
Intervention: Chromatographic separation was achieved using a C18 column (150×4.6 mm, 5 µm) with a mobile phase of Methanol:Acetonitrile (80:20 v/v), a flow rate of 1.0 mL/min, and UV detection at 254 nm.
Main Outcome Measures: The method was evaluated for linearity, accuracy, precision, ruggedness, specificity, and forced degradation under acidic, basic, oxidative, thermal, and wet heat conditions.
Results: Accuracy values ranged between 99.02% and 99.72% for both drugs. %RSD for precision and ruggedness were consistently below 0.32%. Forced degradation showed significant degradation in basic (8.97%) and oxidative (6.25%) conditions, while all degradation products were well-separated from the analyte peaks, confirming specificity.
Conclusion: The developed UPLC method is specific, accurate, and stability-indicating. It meets regulatory validation criteria and is suitable for routine quality control and stability testing of Sulfamethoxazole and Trimethoprim injectable formulations.
Keywords: Sulfamethoxazole, Trimethoprim, UPLC, Method Validation, Forced Degradation, Stability-Indicating Method, ICH Guidelines
Keywords:
Sulfamethoxazole, Trimethoprim, UPLC, Method Validation, Forced degradation, Stability-indicating methodDOI
https://doi.org/10.22270/jddt.v15i6.7224References
1. Ghanem MM, Abu-Lafi SA. Sci Pharm. 2012;81(1):167-182. https://doi.org/10.3797/scipharm.1210-12 PMid:23641336 PMCid:PMC3617661
2. Ascalone V. J Chromatogr B. 1981;224:59-66. https://doi.org/10.1016/S0378-4347(00)80138-3 PMid:7263821
3. Taylor RB, Richards ME, Xing DK. Analyst. 1990;115(6):797-799. https://doi.org/10.1039/an9901500797 PMid:2393083
4. Blessy M, Patel RD, Prajapati PN, Agrawal YK. J Pharm Anal. 2014;4(3):159-165. https://doi.org/10.1016/j.jpha.2013.09.003 PMid:29403878 PMCid:PMC5761119
5. Bakshi M, Singh S. J Pharm Biomed Anal. 2002;28(6):1011-1040. https://doi.org/10.1016/S0731-7085(02)00047-X PMid:12049968
6. Rao BM, Raju AN. J Chromatogr Sci. 2010;48(6):460-464. https://doi.org/10.1007/s00337-010-0809-4
7. Sahu A, Agrawal YK. J Liq Chromatogr Relat Technol. 2015;38(5):539-546.
8. ICH. Validation of analytical procedures: text and methodology Q2(R1). Geneva: ICH; 2005.
9. U.S. FDA. Analytical Procedures and Methods Validation for Drugs and Biologics. 2015.
10. United States Pharmacopeia 42/NF 37. Rockville: USP; 2019.
11. Indian Pharmacopoeia. Ghaziabad: IPC; 2018.
12. The Merck Index. 14th ed. Merck & Co.; 2006.
13. Beckett AH, Stenlake JB. Practical Pharmaceutical Chemistry. 4th ed. CBS; 2007.
14. ICH. Stability testing of new drug substances and products Q1A(R2). Geneva: ICH; 2003.
15. Snyder LR, Kirkland JJ, Dolan JW. Introduction to Modern Liquid Chromatography. 3rd ed. Wiley; 2009. https://doi.org/10.1002/9780470508183
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