N 4 (189) 2024. P. 84–89

VALIDATION OF THE METHOD OF QUANTITATIVE DETERMINATION OF THE ACTIVE SUBSTANCE ((S)-2,6-DIAMINOHEXANOIC ACID 3-METHYL-1,2,4-TRIAZOLYL-5-THIOACETATE IN EYE DROPS

Zaporizhzhia State Medical and Pharmaceutical University, Zaporizhzhia, Ukraine
Zaporizhzhia Research Experimental Forensic Center of the Ministry of Internal Affairs of Ukraine, Zaporizhzhia, Ukraine

DOI 10.32782/2226-2008-2024-4-14

The purpose of the present work is to validate the method of quantitative determination of the active substance ((S)-2,6-diaminohexanoic acid 3-methyl-1,2,4-triazolyl-5-thioacetate) in Angiolin 1% eye drops by the method of high-performance liquid chromatography.

Materials and methods. Certified substances were used: Angiolin, working standard sample, sodium chloride, purified water as auxiliary substances, liquid chromatograph with UV detector; column Hypersil ODS C-18250 X 4.6 with a particle size of 5 μm.

Results. During the determination of validation parameters, it is established that the method is characterized by sufficient correctness, since the criterion of insignificance of the systematic error of the method is fulfilled. The systematic error of the method satisfies the requirements of statistical and practical insignificance. The high value of the correlation coefficient r=0.9999 satisfies the requirements of the acceptance criterion (r=0.99810) and confirms the linearity of the dependence between the amount of Angiolin taken and found in the range from 80% to 120%, following its nominal content in the preparation. The requirements for parameters of linear dependence (a, SD0/b, r) of the Angiolin determination method are met in the entire concentration range from 80% to 120% of the nominal value.

Conclusions. The method of determining Angiolin by HPLC in the range of application of the method meets the acceptance criteria for validation characteristics: specificity, correctness, precision (convergence) and linearity. The total predicted uncertainty of the analyses results does not exceed the critical value regulated by the SPhU, and can be entered into the project of Quality Control Methods.

Key words: eye drops, analysis, quantitative determination, high-performance liquid chromatography, validation.

REFERENCES

1. Belenichev IF, Kucherenko LI, Mazur IA, Akopyan RR, Portnaya EA. Experimental assessment of the specific activity of a new dosage form of Angiolin in cataract modeling. Biological Markers and Guided Therapy. 2019; 6(1) :113–120. Available from: https://doi.org/10.12988/bmgt.2019.91013.
2. Mazur IA, Akopian RR, Cherkovska LH, Pavliuk IV, Skoryna DYu. Development of standardization methods of Angiolin eye drops. Current issues in pharmacy and medicine: science and practice. 2020; 13(3): 383–387. (in Ukrainian). Available from: https://doi.org/10.14739/2409-2932.2020.3.216214.
3. Kucherenko LI, Mazur IA, Akopian RR, Portna OO, Tkachenko HI. Scientific substantiation of the concentration of excipients for the manufacture of eye drops Angiolin. Current issues in pharmacy and medicine: science and practice. 2021; 14(1): 52–55. (in Ukrainian). Available from: https://doi.org/10.14739/2409-2932.2021.1.226773.
4. Grizodub AI. Standardization of the procedure for validation of methods of quality controlof drugs. State Enterprise “Ukrainian Scientific Pharmacopoeial Center for Drug Quality”; 2016. 396 р. (in Russian).
5. Georgievskii VP, Georgievskii GV, Zinchenko AA, Kulikov AYu, Nazarova ES, Kolisnyk AV. Chromatographic methods in analytical support for the creation and quality control of medicines in Ukraine. Kharkiv: NTMT; 2016. 288 р. (in Russian).
6. State Enterprise Ukrainian Scientific Pharmacopoeial Center of Medicines Quality. 2nd ed. Kharkiv: State Enterprise Ukrainian Scientific Pharmacopoeial Center of Medicines Quality. 2015. 1126 p. (in Ukrainian).