Spectrophotometric Determination of Doxycycline Hydrochloride by Diazotization - Coupling Using Ortho-Anisidine Reagent Application to Pharmaceutical Formulations

Year 2025, Volume: 10 Issue: 2, 169 - 183, 01.09.2025

Karam Sh. Shakir , Dawood H. Mohammed

https://doi.org/10.28978/nesciences.1698459

Abstract

The following project aims to produce a sensitive, high-precision and fast spectroscopic method. and cost-effective estimate for the determination of doxycycline hydrochloride in pharmaceuticals preparations), the reaction of Diazotization and coupling , The proposed approach is based on the creation of dyzonium salt by the reaction of In an acidic media, the ortho-anisidine reagent is combined with nitrite before being coupled with doxycycline hydrochloride in an alkaline medium to create a Yellow dye with the highest absorption at 437 nm , a calibration curve within Beer's law and concentration (1-60 μg.ml-1) with a correlation coefficient of 0.9998 of doxycycline hydrochloride, the detection limits were 0.379 (LOD) and 1.26 (LOQ), the Sandell’s sensitivity was 0.044 µg.cm-2, and the molar absorptivity was 10.72 x 103 l/mol.cm, relative standard deviation whose value not exceed 0.430 % and the recovery value not less than 99.96%, The method has been applied to pharmaceutical products. in several forms of doxycycline showed that the proposed method has good results within the acceptable error, The validity of the proposed method was proved by a recovery investigation Using the standard addition method. the validated approach is appropriate and extremely helpful for routine quality control of doxycycline hydrochloride since it does not require polluting reagents and is quick and inexpensive.

Keywords

Doxycycline hydrochloride , Ortho-anisidine , Determination , spectrophotometric , Diazotization and coupling

References

• Al-Abachi, M. Q., & Al-Nedawi, Z. A. (2015). Batch and flow injection spectrophotometric determination of doxycycline hyclate in pharmaceutical preparations. Al-Nahrain Journal of Science, 18(3), 24-32.
• Alassaf, N. A., & Faeza, H. Z. (2019). Sensitive spectrophoto metr doxycycline in pure and Dos p Bromanil. International Journal of Pharmaceutical Research, 11(2). 10.31838/ijpr/2019.11.02.016
• Ali, T. A., Mohamed, G. G., El-Sonbati, A. Z., Diab, M. A., & Elkfass, A. M. (2018). A potentiometric sensor for determination of doxycycline hydrochloride in pharmaceutical preparation and biological fluids. Russian Journal of Electrochemistry, 54(12), 1081-1095. https://doi.org/10.1134/S1023193518120029
• Amit, K., Sanjo, N., & Rajive, C. (2010). Spectrophotometeric Methods for Determination of Doxycycline in Tablet Formulation. International Journal of PharmTech Research, 2, 599-602.
• Bhattacharya, S. K. (2003). An evaluation of current cholera treatment. Expert opinion on pharmacotherapy, 4(2), 141-146.https://doi.org/10.1517/14656566.4.2.141
• Chen, C. J., Gillett, A., Booth, R., Kimble, B., & Govendir, M. (2022). Pharmacokinetic profile of doxycycline in koala plasma after weekly subcutaneous injections for the treatment of chlamydiosis. Animals, 12(3), 250. https://doi.org/10.3390/ani12030250
• Chen, Y. F., Yang, Y. N., Chu, H. R., Huang, T. Y., Wang, S. H., Chen, H. Y., ... & Davis, P. J. (2022). Role of integrin αvβ3 in doxycycline-induced anti-proliferation in breast cancer cells. Frontiers in cell and developmental biology, 10, 829788. https://doi.org/10.3389/fcell.2022.829788
• Chinnasamy. (2024). A Blockchain and Machine Learning Integrated Hybrid System for Drug Supply Chain Management for the Smart Pharmaceutical Industry. Clinical Journal for Medicine, Health and Pharmacy, 2(2), 29-40.
• Delevie, R. (1997). Principles of quantitative chemical analysis. Singapore: McGraw-Hill International Edition.
• Dil, E. A., Ghaedi, M., Asfaram, A., Tayebi, L., & Mehrabi, F. (2020). A ferrofluidic hydrophobic deep eutectic solvent for the extraction of doxycycline from urine, blood plasma and milk samples prior to its determination by high-performance liquid chromatography-ultraviolet. Journal of Chromatography a, 1613, 460695. https://doi.org/10.1016/j.chroma.2019.460695
• Feng, X., Ashley, J., Zhou, T., & Sun, Y. (2018). Fluorometric determination of doxycycline based on the use of carbon quantum dots incorporated into a molecularly imprinted polymer. Microchimica Acta, 185(11), 500. https://doi.org/10.1007/s00604-018-2999-8
• Foroutan, B., Bashi Amlashi, H. R., Partani, A., De los Ríos, P., & Nasrollahzadeh Saravi, H. (2023). Determination and comparisons of heavy metals (Cobalt and Iron) accumulation in muscle, liver, and gill tissues of Golden Mullet (Chelon aurata) in coastal areas of the Caspian Sea (Mazandaran and Golestan provinces of Iran). International Journal of Aquatic Research and Environmental Studies, 3(1), 1-15. https://doi.org/10.70102/IJARES/V3I1/1
• Ghidini, L., Kogawa, A., & Salgado, H. R. N. (2018). Eco-friendly green liquid chromatographic for determination of doxycycline in tablets and in the presence of its degradation products. Drug Analytical Research, 2(2), 49-55. https://doi.org/10.22456/2527-2616.89412
• Gürler, B., Özkorucuklu, S. P., & Kır, E. (2013). Voltammetric behavior and determination of doxycycline in pharmaceuticals at molecularly imprinted and non-imprinted overoxidized polypyrrole electrodes. Journal of pharmaceutical and biomedical analysis, 84, 263-268. https://doi.org/10.1016/j.jpba.2013.06.009
• Hargis, D. C. (2016). Quantitative chemical analysis (9th ed.). New York, NY: W. H. Freeman and Company.
• Hashim, H. A., Maulood, M. F., Rasheed, A. M., Fatak, D. F., Kabah, K. K., & Abdulamir, A. S. (2020). Controlled randomized clinical trial on using Ivermectin with Doxycycline for treating COVID-19 patients in Baghdad, Iraq. MedRxiv, 2020-10.
• Khammas, Z. A., & Rashid, R. A. (2016). Visible spectrophotometric analysis for the mutual determination of doxycycline hydrochloride and iron in real samples after cloud point extraction. International Journal of Chemical Science, 14(2), 955-977.
• Kogawa, C., & Salgado, H. R. N. (2012). Doxycycline hyclate: A review of properties, applications and analytical methods. International Journal of Life Science and Pharmaceutical Research, 2(4), 11–25.
• Korolkovas, A., & Burckhalter, J. H. (1981). Química Farmacêutica, Editora Guanabara Koogan. SA, Rio de Janeiro.
• Kumssa, L., Layloff, T., Hymete, A., & Ashenef, A. (2021). High performance thin layer chromatography (HPTLC) method development and validation for determination of doxycycline hyclate in capsule and tablet formulations. Acta Chromatographica. DOI: https://doi.org/10.1556/1326.2021.00926
• Mashru, R., & Koshti, N. (2021). Development and validation of UV-Spectrophotometric and RP-HPLC method for simultaneous estimation of Metformin and Doxycycline in bulk and synthetic mixture. Journal of Drug Delivery and Therapeutics, 11(4-S), 26-35.
• Milan, T., & Žarko, N. (2018). Determination of Air Refraction Influence on Trigonometric Height Differences. Archives for Technical Sciences/Arhiv za Tehnicke Nauke, (18).
• Mileva, R. (2019). Determination of free doxycycline concentrations in the plasma and milk of sheep and in the plasma of rabbits by using the HPLC method. Macedonian Veterinary Review, 42(2). https://doi.org/10.2478/macvetrev-2019-0016
• Palamy, S., & Ruengsitagoon, W. (2017). A novel flow injection spectrophotometric method using plant extracts as green reagent for the determination of doxycycline. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 171, 200-206. https://doi.org/10.1016/j.saa.2016.08.011.
• Permana, A. D., Tekko, I. A., McCarthy, H. O., & Donnelly, R. F. (2019). New HPLC–MS method for rapid and simultaneous quantification of doxycycline, diethylcarbamazine and albendazole metabolites in rat plasma and organs after concomitant oral administration. Journal of Pharmaceutical and Biomedical Analysis, 170, 243-253. https://doi.org/10.1016/j.jpba.2019.03.047
• Phaechamud, T., & Charoenteeraboon, J. (2008). Antibacterial activity and drug release of chitosan sponge containing doxycycline hyclate. Aaps PharmSciTech, 9(3), 829-835.
• Prakash, J., & Meena, K. (2022). Simulation of Anti Reflecting Coating for Improving External Quantum Efficiency of Photovoltaic Cell. Journal of Energy Engineering and Thermodynamics, 2(04), 24-35. https://doi.org/10.9756/IAJSE/V9I1/IAJSE0905
• Ramesh, P. J., Basavaiah, K., Divya, M. R., Rajendraprasad, N., Vinay, K. B., & Revanasiddappa, H. D. (2011). Simple UV and visible spectrophotometric methods for the determination of doxycycline hyclate in pharmaceuticals. Journal of Analytical chemistry, 66(5), 482-489. https://doi.org/10.1134/S1061934811050157.
• Ramesh, P. J., Basavaiah, K., Tharpa, K., Vinay, K. B., & Revanasiddappa, H. D. (2010). Development and validation of RP-HPLC method for the determination of doxycycline hyclate in spiked human urine and pharmaceuticals. Journal of Pre-Clinical and Clinical Research, 4(2).
• Rufino, J. L., Weinert, P. L., Pezza, H. R., & Pezza, L. (2009). Flow-injection spectrophotometric determination of tetracycline and doxycycline in pharmaceutical formulations using chloramine-T as oxidizing agent. Química Nova, 32, 1764-1769. https://doi.org/10.1590/S0100-40422009000700016
• Ruiz, S. M. A., Olvera, L. G., Bernad, M. J. B., Chacón, S. D. C. C., & Estrada, D. V. (2015). Comparative pharmacokinetics of a new oral long-acting formulation of doxycycline hyclate: A canine clinical trial. European Journal of Pharmaceutical Sciences, 80, 9-15. https://doi.org/10.1016/j.ejps.2015.09.012
• Saber, A. L., & Amin, A. S. (2011). Utility of Ion-Pair and Charge Transfer Complexation for Spectrophotometric Determination of domperidone and doxycycline in bulk and pharmaceutical formulations. J Anal Bioanal Techniques, 1(113), 2. 10.4172/2155-9872.1000113
• Sancho, E., Granados-Chinchilla, F., & Barquero-Calvo, E. (2022). Determination of streptomycin and doxycycline using LC/MS towards an effective treatment against an experimental Brucella abortus infection in mice. Journal of Microbiological Methods, 194, 106436. https://doi.org/10.1016/j.mimet.2022.106436
• Sheeja, N. K. (2019). Open Access Initiatives of INFLIBNET: An Analytical Study. Indian Journal of Information Sources and Services, 9(2), 20–23. https://doi.org/10.51983/ijiss.2019.9.2.631
• Singh, C., & Gurudiwan, P. (2024). Design and Modeling of Sustainable Environment in Pharmacy and Pharmaceutical Practices. Natural and Engineering Sciences, 9(2), 449-459.http://doi.org/10.28978/nesciences.1575487
• Skoog, D. A., West, D. M., Holler, F. J., & Crouch, S. R. (1996). Fundamentals of analytical chemistry (Vol. 33, pp. 53-55). Fort Worth: Saunders College Pub.
• Tan, K. R., Magill, A. J., Parise, M. E., & Arguin, P. M. (2011). Doxycycline for malaria chemoprophylaxis and treatment: report from the CDC expert meeting on malaria chemoprophylaxis. The American journal of tropical medicine and hygiene, 84(4), 517.https://doi.org/10.4269/ajtmh.2011.10-0285
• Tawfeeq, A. H., & Qassim, B. B. (2020). A green method for assay of doxycycline hyclate using continuous flow injection/merging zones technique via coupling with azo metol in aqueous medium. Voprosy khimii i khimicheskoi tekhnologii, 4, 31-37. DOI: 10.32434/0321-4095-2020-131-4-31-37
• The Stationery Office. (2020). British Pharmacopoeia (CD-ROM ed., pp. 852–853). London: The Stationery Office.
• Tian, X., & Fan, Z. (2021). Novel ratiometric probe based on the use of rare earth-carbon dots nanocomposite for the visual determination of doxycycline. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 260, 119925. https://doi.org/10.1016/j.saa.2021.119925
• Verma, A., & Nair, R. (2025). Chromatographic Methods for the Separation of Naturally Occurring Bioactive Compounds and Their Applications in Industry. Engineering Perspectives in Filtration and Separation, 18-24. https://filtrationjournal.com/index.php/epfs/article/view/EPFS25104
• Zhuang, Y., Lin, B., Yu, Y., Wang, Y., Zhang, L., Cao, Y., & Guo, M. (2021). A ratiometric fluorescent probe based on sulfur quantum dots and calcium ion for sensitive and visual detection of doxycycline in food. Food Chemistry, 356, 129720. https://doi.org/10.1016/j.foodchem.2021.129720.