Validation of an analytical method for the quantification of total alkaloids
Keywords:
belladonna alkaloids, pharmaceutical industry, validation studies.Abstract
Introduction: The validation of analytical methods ensures the reproducibility of data, when it is desired to perform the quantification of a product, using methodologies different from those established in official documents.Objective: Validate an analytical method by UV Vis spectrophotometry for the quantification of total alkaloids.
Methods: A galenic preparation containing 0.1% of belladonna extract was used to determine the validation parameters using a UV-visible spectrophotometric method at 435 nm, based on the precipitation of alkaloids by interaction with the Dragendorff's reagent. Specificity used a concentration of 100 %, while linearity required samples of 80 %, 90 %, 100 %, 110 % and 120 %; and, in the case of the range, at 80 % and 120 %. Precision evaluated repeatability at 80 %, 100 % and 120 %, and intermediate precision required a 100 % sample considering inter-analyst, inter-team and inter-day conditions. Finally, in accuracy, concentrations of 80 %, 100% and 120 %; and, for robustness, a 100 % sample was analyzed to determine pH variation and centrifugation speed.
Results: The selectivity obtained acceptable values in both the matrix and the elaborated product, the linearity presented correlation and determination coefficients close to 1, and the concentration range was between 40 - 60 mg/L. Accuracy, precision and robustness had a variability of less than 2 %.
Conclusions: The UV Vis spectrophotometric method is selective, linear, exact, precise and robust; it is reliable in the determination of precipitable alkaloids.
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References
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5. Zhang Z, Li Y, Sun Y, Wang W, Song X, Zhang D. Chemical diversity and biological activities of marine-derived sulphur containing alkaloids: A comprehensive update. Arab J Chem. 2023; 16(9):105011. DOI: 10.1016/j.arabjc.2023.105011
6. Rajput A, Sharma R, Bharti R. Pharmacological activities and toxicities of alkaloids on human health. Mater Today Proc. 2022; 48:1407-15. DOI: 10.1016/j.matpr.2021.09.189
7. Efferth T, Oesch F. Repurposing of plant alkaloids for cancer therapy: Pharmacology and toxicology. Semin Cancer Biol. 2021; 68:143-63. DOI: 10.1016/j.semcancer.2019.12.010
8. Yang M, Ma J, Ruan J, Zhang C, Ye Y, Pi-Cheng Fu P, et al. Absorption difference between hepatotoxic pyrrolizidine alkaloids and their N-oxides - Mechanism and its potential toxic impact. J Ethnopharmacol. 2020; 249:112421. DOI: 10.1016/j.jep.2019.112421
9. Sattler M, Müller V, Bunzel D, Kulling SE, Soukup ST. Pyrrolizidine alkaloids in borage (Borago officinalis): Comprehensive profiling and development of a validated LC-MS/MS method for quantification. Talanta. 2023; 258:124425. DOI: 10.1016/j.talanta.2023.124425
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11. Suselo YH, Indarto D, Wasita B, Hartono H. Alkaloid fraction of Mirabilis jalapa Linn. flowers has low cytotoxicity and increases iron absorption through Erythropoietin-Matriptase-2-Hepcidin pathway in iron deficiency Hepatocarcinoma cell model. Saudi J Biol Sci. 2023; 30(1):103508. DOI: 10.1016/j.sjbs.2022.103508
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19. The Unites States Convention. Farmacopea de los Estados Unidos de América: Formulario Nacional, Compendios de normas oficiales. Validación de procedimientos farmacopeicos. USP 44 - NF 39. EE. UU., Rockville: U.S. Pharmacopeial Convention, Inc. 2022, pp: 8387-8393.
20. Versilovskis A, Mulder P, Pereboom D, Stoppelaar J, Nijs M. Simultaneous quantification of ergot and tropane alkaloids in bread in the Netherlands by LC-MS/MS. Food Addit Contam 2020; 13(3): 215-23. DOI: 10.1080/19393210.2020.1771777
21. Eugelio F, Palmieri S, Fanti f, Messuri L, Pepe A, Compagnone D, et al. Development of an HPLC-MS/MS method for the determination of alkaloids in lupins. Molecules 2023; 28(4): 1531. DOI: 10.3390/molecules28041531
Published
2024-05-10
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Zavaleta Juárez ME, Castillo Saavedra EF, La Cunza Méndez AD, Noriega Oblitas JM, Reyes Alfaro CE. Validation of an analytical method for the quantification of total alkaloids. Rev Cubana Med Milit [Internet]. 2024 May 10 [cited 2025 Apr. 3];53(2):e024025549. Available from: https://revmedmilitar.sld.cu/index.php/mil/article/view/25549
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