Development and validation of an ecological, new and rapid stability-indicating High Performance Liquid Chromatographic method for quantitative determination of aztreonam in lyophilized powder for injection
DOI:
https://doi.org/10.22456/2527-2616.73755Keywords:
aztreonam, analytical methods, HPLC, quality control, validationAbstract
Aztreonam is a monocyclic synthetic antimicrobial with bactericidal activity against Gram-negative bacteria, the first agent from the monobactam family to be therapeutically approved. It was developed and validated analytical method using high performance liquid chromatography with UV detection at 292 nm to quantify the aztreonam. Furthermore, assessing stability through stress tests was carried out. The chromatographic separation was carried out by reverse phase on an Agilent C18 column (250 x 4.6 mm, 5 µm) with a mobile phase composed of water:ethanol (70:30, v/v) adjusted to pH 2.5 with acetic acid, pumped isocratically at a flow rate of 0.5 mL/min. The validation parameters linearity, selectivity, precision, accuracy, robustness, limits of detection and quantification were determined. The method proposed provides linear response within the concentration range of 45-95 µg/mL for aztreonam. Results obtained were found to be satisfactory. The proposed method is linear, accurate, precise, selective, and robust being able to quantify the aztreonam in raw material and in pharmaceutical preparations. The validated method was suitable for applications in quality control laboratories.
Downloads
References
Srinivas, N.R., Papp, E.A., Shah, V.R., Shyo, W.C. A simple HPLC assay, with ultraviolet detection, for determination of a monobactam antibiotic. J Liquid Chrom & Rel Technol. 1997; 20:1091-1101. DOI: 10.1080/10826079708010961. DOI: https://doi.org/10.1080/10826079708010961
US Pharmacopeia 37th, US Pharmacopeial Convention, Rockville, 2014.
Georgopapadakou N.H., Smith S.A. and Sykes R.B. Mode of action of azthreonam. J Pharm Biomed Anal, 1982;21:950–956. DOI: https://doi.org/10.1128/AAC.21.6.950
Axelrod, J., Daly, J.S., Barza, M., Baker, A.S. Antibacterials. In: Albert DM, Jakobiec FA, Ed. Principles and practice of ophthalmology, Philadelphia, W.B. Philadelphia; p.940-961, 1994.
Loeb, S. Physician’s drug handbook, 5th ed. Springhouse Corporation: Springhouse; 1993.
Papich, M.G., Ettinger, S.J., Feldman, E.C. Antimicrobial drugs. Textbook of veterinary internal medicine, 4th edn WB Saunders Company: Philadelphia; 1:272-274, 1995.
Sykes, R.B., Koster, W.H., Bonner, D.P. The new monobactams: chemistry and biology. J. Clin. Pharmacol. 1988; 28:113–119. DOI: https://doi.org/10.1002/j.1552-4604.1988.tb05734.x
Kapoor, S., Gathwala, G. Aztreonam. Indian Pediatrics, 2004;41:359–364.
Giamarellou, H., Galanakis, N., Douzinas, E., Petrikkos, G., El Messidi, M., Papoulias, G., Daikos, G.K. Evaluation of aztreonam in difficult to treat infections with prolonged post-treatment follow up. Antimicrob Agents Chemother. 1984;26:245-249. DOI: https://doi.org/10.1128/AAC.26.2.245
Swabb, E.A., Sugerman, A.A., Stern, M. Oral bioavailability of the monobactam aztreonam (SQ 26,776) in healthy subjects. Antimicrob Agents Chemother. 1983; 23:548-550. DOI: https://doi.org/10.1128/AAC.23.4.548
Swabb, E.A. Review of the clinical pharmacology of the monobactam antibiotic aztreonam. Am J Med. 78:11-18, 1985. DOI: https://doi.org/10.1016/0002-9343(85)90197-4
Davis, J.D. Aztreonam, Elsevier Science: New York; 4:61-64, 1997. DOI: https://doi.org/10.1016/S1068-607X(96)00066-2
Zajac, M., Jelinska, A., Cielecka-Piontek, J., Oszczapowicz, I. Stability of aztreonam in AZACTAM. II Farmaco, 2005; 60:599-603. DOI 10.1016/j.farmac.2005.04.009. DOI: https://doi.org/10.1016/j.farmac.2005.04.009
Pilkiewick, F.G., Remsburg, B.J., Fisher, S.M., Sykes, R.B. High-pressure liquid chromatographic analysis of aztreonam in serum and urine. Antimicrob Agents Chemother. 1983;23:852-856. DOI: https://doi.org/10.1128/AAC.23.6.852
Egger, H., Fischer, G. Determination of the monocyclic beta-lactam antibiotic carumonam in plasma and urine by ion-pair and ion-suppression reversed-phase high-performance liquid chromatography. J Chromatography, 1987;420:357- 372. DOI: https://doi.org/10.1016/0378-4347(87)80191-3
Ranadive, S.A., Pipkin, J.D., Varia, S.A., Chang, N.H., Barry, E.P., Porubcan, M., et al. Formation, isolation and identification of oligomers of aztreonam. Eur J Pharm Sci, 1995;3:281-291. DOI: https://doi.org/10.1016/0928-0987(95)00017-8
ICH Q2 (R1). Validation of analytical procedures: text and methodology, in Proceedings of the International Conference on Harmonization, IFPMA, Geneva, 2005.
Brasil, Agência Nacional de Vigilância Sanitária, RE 899, Guia para validação de métodos analíticos e bioanalíticos, Brasília, 2003.
Zivanovic, L., Zigic, G., Zecevic, M. Investigation of chromatographic conditions for the separation of ofloxacin and its degradation products. J Chromatogr A, 2006; 1119:224-230. DOI: https://doi.org/10.1016/j.chroma.2006.02.029
Lopes, C.C.G.O., Salgado, H.R.N. Development of a validated stability-indicating LC assay and stress degradation studies of linezolid in tablets. Chromatographia, 2009; 69:S129–S135. DOI 10.1365/s10337-009-0993-3. DOI: https://doi.org/10.1365/s10337-009-0993-3
Cazedey, E.C.L., Perez, D.P., Perez, J.P., Salgado, H.R.N. A high performance liquid chromatographic assay for ciprofloxacin hydrochloride ophthalmic solution. Chromatographia, 2009; 69:S241–S244. DOI 10.1365/s10337-009-1004-4. DOI: https://doi.org/10.1365/s10337-009-1004-4
FDA, Validation of chromatographic methods, Center of Drug Evaluation and Research: Washington; 2004.
Inmetro DOQ-CGCRE-008, Orientação sobre validação de métodos analíticos, 2011.
Brazilian Pharmacopeia, ANVISA: Brasília; 5th edn, 2010.
Joshi, S. HPLC separation of antibiotics present in formulated and unformulated samples. J Pharm Biomed Anal, 2002; 28:795-809. DOI: https://doi.org/10.1016/S0731-7085(01)00706-3
Karcher, B.D., Davies, M.L., Delaney, E.J., Vent, J.J. A 21st Century HPLC workflow for process R&D. JALA, 2005; 381-393. DOI: https://doi.org/10.1016/j.jala.2005.09.006
Rugani, K.S., Salgado, H.R.N. Stability-indicating LC method for the determination of cephalothin in lyophilized powder for injection. Anal Meth, 2014; 6:4437-4445. DOI: 10.1039/C3AY42049C. DOI: https://doi.org/10.1039/C3AY42049C
Bonfilio, R., Cazedey, E.C.L., Araújo, M.B., Salgado, H.R.N. Analytical validation of quantitative high-performance liquid chromatographic methods in pharmaceutical analysis: A practical approach. Crit Rev Anal Chem, 2012;42:87–100. http://dx.doi.org/10.1080/10408347.2012.630926 DOI: https://doi.org/10.1080/10408347.2012.630926
Pedroso, T.M., Medeiros, A.C.D., Salgado, H.R.N. RP-HPLC x HILIC chromatography for quantifying the ertapenem sodium with a look at green chemistry. Talanta, 2016; 160:745–753. http://dx.doi.org/10.1016/j.talanta.2016.08.016i
Gałuszka, A., Migaszewski, Z., Namies’nik, J. The 12 principles of green analytical chemistry and the SIGNIFICANCE mnemonic of green analytical practices. Trends Anal Chem. 2013; 50:78–84. DOI: https://doi.org/10.1016/j.trac.2013.04.010
Salgado, H.R.N. New analytical methods for a new awake society. EC Microbiol. 2017; 9(2)39.
Ribeiro, R.L.V., Bottoli, C.B.G., Collins, K.E.,Collins, C.H. Reevaluation of ethanol as organic modifier for use in HPLC-RP mobile phases. J Braz Chem Soc. 2004; 15(2):300-306. DOI: https://doi.org/10.1590/S0103-50532004000200022
Ribeiro, R.L.V., Grespan, C.B., Collins, C.H., Collins, K.E., Bruns, R.E. Optimization through factorial planning of the use of ethanol:water as a mobile phase for reversed phase HPLC. J High Resolut Chromatogr. 1999; 22(1): 52-54. DOI: https://doi.org/10.1002/(SICI)1521-4168(19990101)22:1<52::AID-JHRC52>3.0.CO;2-T
Miyabe, K., Takeuchi, S., Tezuka, Y. Adsorption characteristics in reversed-phase liquid chromatography using ethanol/water mixed solvent. Adsorption, 1999; 5:15-24. DOI: https://doi.org/10.1023/A:1026477619947
Moreno, A.H., Salgado, H.R.N. Development and validation of HPLC method for determination of ceftazidime. J AOAC Int., 2008; 91:739-743. DOI: https://doi.org/10.1093/jaoac/91.4.739
Vieira, D.C.M., Salgado, H.R.N. Comparison of HPLC and UV spectrophotometric methods for the determination of cefuroxime sodium in pharmaceutical products. J Chromatogr Sci. 2011; 49:508-511. DOI: https://doi.org/10.1093/chrsci/49.7.508
Tótoli, E.G., Salgado, H.R.N. Development and validation of an economic, environmental friendly and stability-indicating analytical method for determination of ampicillin sodium for injection by RP-HPLC. World J Pharm Pharmac Sci. 2014; 3(6): 1928-1943.
Tótoli, E.G., Salgado, H.R.N. Development and validation of an economic, environmental friendly and stability-indicating analytical method for determination of ampicillin sodium for injection by RP-HPLC. World J Pharm Pharmac Sci. 2014; 3:1928-1943.
Tótoli, E.G., Salgado, H.R.N. Development, optimization and validation of a green and stabilityindicating HPLC method for determination of daptomycin in lyophilized powder. J AOAC Int. 2015; 98:1276-1285. DOI: https://doi.org/10.5740/jaoacint.15-039
Pedroso, T.M., Medeiros, A.C.D., Salgado, H.R.N. RP-HPLC x HILIC chromatography for quantifying the ertapenem sodium with a look at green chemistry. Talanta, 2016; 160:745–753. http://dx.doi.org/10.1016/j.talanta.2016.08.016i DOI: https://doi.org/10.1016/j.talanta.2016.08.016
Rodrigues, D.F., Salgado, H.R.N. Development and validation of a green analytical method of RPHPLC for quantification cefepime hydrochloride in pharmaceutical dosage forms: simple, sensitive and economic. Cur Pharm Anal. 2016; 12:306-314. Doi 10.2174/1573412912666151221210921. DOI: https://doi.org/10.2174/1573412912666151221210921
Figueiredo, A.L., Salgado, H.R.N. Validation of a green analytical method for the quantitative analysis of antimicrobial aztreonam in lyophilized powder for injection by Fourier-Transform Infrared Spectroscopy (FT-IR). EC Microbiol. 2017; 8(5):254-265. https://www.ecronicon.com/ecmi/pdf/ECMI-08- 00257.pdf
Figueiredo, A.L., Salgado, H.R.N. Development and validation of a ultraviolet (UV) spectrophotometric method for determination of aztreonam in pharmaceutical products. EC Microbiol. 2017; 8(6):305-316.
Downloads
Published
Versions
- 10-03-2026 (5)
- 07-03-2026 (4)
- 06-03-2026 (3)
- 28-08-2017 (2)
- 28-08-2017 (1)
How to Cite
Issue
Section
License
Copyright (c) 2017 Drug Analytical Research

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
The Copyright holder of manuscripts published is Drug Analytical Research. Authors who publish with this journal are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
Licensing:
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License
Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.





