The investigations of propranolol by HPLC method suitable for a chemical-toxicological analysis
Topicality. Therapeutic monitoring when using propranolol hydrochloride, analysis of the drug in dosage forms and biological objects, identification and quantitative determination of the drug during intoxication are carried out using sensitive and selective methods. Among the modern methods of analysis for creating a database of parameters of identification and quantitative determination of analytes arrays in biological objects HPLC method is one of the most suitable methods with high sensitivity and selectivity.
Propranolol hydrochloride (anaprilin) - (±)1-isopropylamino-3-(1-naphthyloxy) -2-propanol hydrochloride – non-selective β-adrenoceptor blocker, which is characterized by antianginal, antihypertensive and antiarrhythmic effects and is used for treatment of coronary heart disease, heart rhythm disorders and some forms of hypertension. The previously developed HPLC methods of propranolol analysis are based on the application of various chromatographic conditions (mobile phase composition, isocratic or gradient elution, detecting at one or several wavelengths, selection of sensitive and selective detector) due to individual properties of a substance. Taking into account the possibility of complex treatment of diseases of the cardiovascular system with the use of different drugs, the analysis by a unified HPLC methodology is an actual task of the study.
The purpose of this work is identification and quantitative determination of propranolol hydrochloride by a unified HPLC methodology, which allows obtaining reliable and reproducible research results that are suitable for drug analysis in biological objects.
Materials and methods of research. Chromatography of the investigated substance was performed on a microcolonial liquid chromatograph "Milichrome A-02" ("EcoNova", Novosibirsk, Russia) in an reverse-phase variant using a metal column with a nonpolar sorbent Prontosil 120-5C 18 AQ, 5 μm. For elution of the drug were used a mixture of solvents - acetonitrile with 0,2 M solution of lithium perchlorate in 0,005 M solution of acid perchloric. The linear gradient from eluent A (5% acetonitrile and 95% buffer solution) to eluent B (100% acetonitrile) for 40 minutes created conditions for the exit from the column of all component parts of the sample as narrow zones. The flow rate of the mobile phase has been formed 100 μl/min, column temperature – 37 - 40 °С; pump pressure – 2,8 – 3,2 MPa; injection volume – 4 μl. Multi-channel detection of a substance was performed by a UV detector in 8 wavelengths: 210, 220, 230, 240, 250, 260, 280 and 300 nm.
Results and their discussion. In carrying out identification of propranolol were established absolute parameters of retention time (19,40-19,54 min) and retention volume (1940,8-1955,2 μl), spectral relationships, detection limit of the substance in the sample (4,0 μg / ml or 16,0 ng in the sample), the values of the coefficients of the symmetry of the peaks of the substance (0,94-1,10) and the coefficients of capacity ratio (11,94-12,04).
The regression coefficients of the calibration graph, which corresponds to the equation of the straight line S = 0,00565 С were calculated by the least squares method. The correlation coefficient was equal to 0,9987. Validation characteristics of HPLC-method for determination of propranolol: linearity range (5,0-100,0 μg / ml), limit of quantitative determination (5,0 μg / ml or 20 ng in the sample), correctness and accuracy on the basis of the results of the quantitative determination of the preparation by the HPLC method in model solutions were calculated. It was established that the relative uncertainty of the average result did not exceed ± 2,11% when using the proposed method of HPLC analysis of propranolol hydrochloride in model solutions.
Conclusions. Identification and quantitative determination of propranolol hydrochloride with the use of a unified HPLC method suitable for a chemical toxicological study was carried out.
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Mashkovskiy, M. D. (2012). Lekarstvennyye sredstva. Moscow: Novaya Volna, 1216.
Amaya, C. N., Perkins, M., Belmont, A., Herrera, C., Nasrazadani, A. (2018). Non-selective beta blockers inhibit angiosarcoma cell viability and
increase progression free- and overall-survival in patients diagnosed with metastatic angiosarcoma. Oncoscience, 5 (3–4), 109–119. https://
Brohée, L., Peulen, O., Nusgens, B., Castronovo, V., Thiry, M., Colige, A. C., & Deroanne, C. F. (2018). Propranolol sensitizes prostate cancer cells to
glucose metabolism inhibition and prevents cancer progression. Scientific Reports, 8(1). https://doi.org/10.1038/s41598-018-25340-9
Vivas-Colmenares, G. V., Bernabeu-Wittel, J., Alonso-Arroyo, V., Matute de Cardenas, J. A., Fernandez-Pineda, I. (2015). Effectiveness of Propranolol in the
Treatment of Infantile Hemangioma Beyond the Proliferation Phase. Pediatric Dermatology, 32(3), 348–352. https://doi.org/10.1111/pde.12520
Jianguo, K., Xichun, W., Mingkun, Z., Qiaoling, C., & Wenjin, L. (2015). Large Doses of Propranolol for the Treatment of Infantile Cephalic and Facial Hemangiomas:
A Clinical Report of 38 Cases. Journal of Clinical Pediatric Dentistry, 39(3), 268–271. https://doi.org/10.17796/1053-4628-39.3.268
Jovic-Stosic, J., Gligic, B., Putic, V., Brajkovic, G., & Spasic, R. (2011). Severe propranolol and ethanol overdose with wide complex tachycardia
treated with intravenous lipid emulsion: A case report. Clinical Toxicology, 49(5), 426–430. https://doi.org/10.3109/15563650.2011.583251
Garg, A., Panda, S., Dalvi, P., Mehra, S., Ray, S., & Singh, V. (2014). Severe suicidal digoxin and propranolol toxicity with insulin overdose. Indian
Journal of Critical Care Medicine, 18(3), 173–175. https://doi.org/10.4103/0972-5229.128709
Clarke, E. J. C. (2011). Isolation and Identification of Drugs in Pharmaceuticals. Body Fluids and Postmortem Material. London: The Pharm. Press, 2463.
Silva Gracia, M., Köppl, A., Unholzer, S., & Haen, E. (2017). Development and validation of an HPLC-UV method for the simultaneous determination
of the antipsychotics clozapine, olanzapine and quetiapine, several beta-blockers and their metabolites. Biomedical Chromatography, 31(10),
Saleem, K., Ali, I., Kulsum, U., & Aboul-Enein, H. Y. (2013). Recent Developments in HPLC Analysis of β-Blockers in Biological Samples. Journal of
Chromatographic Science, 51(8), 807–818. https://doi.org/10.1093/chromsci/bmt030
Kokorina, N. O., Novoselov, V. P., Khanina, M. A. (2008). Sibirskii meditcinskii zhurnal (Tomsk), 2 (4), 51–53.
Baram, G. I. (2005). Khromatograf “Milikhrom A–02“. Opredelenie veshchestv s primeneniem baz dannykh «VEZhKh–UF». Novosibirsk : ZAO Institut khromatografii, 64.
GOST Style Citations
1. Машковский, М. Д. Лекарственные средства : 16–е изд., перераб., испр. и доп. / М. Д. Машковский. – М. : Новая Волна, 2012. – 1216 с.
2. Non-selective beta blockers inhibit angiosarcoma cell viability and increase progression free- and overall-survival in patients diagnosed with metastatic angiosarcoma / C. N. Amaya [et al.] // Oncoscience. – 2018. – Vol. 5, № 3–4. – P. 109–119. https://doi.org/10.18632/oncoscience.413
3. Propranolol sensitizes prostate cancer cells to glucose metabolism inhibition and prevents cancer progression / L. Brohée [et al.] // Sci Rep. – 2018. – Vol. 8, № 1. – P. 7050. https://doi.org/10.1038/s41598-018-25340-9
4. Effectiveness of propranolol in the treatment of infantile hemangioma beyond the proliferation phase / G. V. Vivas-Colmenares [et al.] // Pediatr Dermatol. – 2015. – Vol. 32, № 3. – P. 348–352. https://doi.org/10.1111/pde.12520
5. Large Doses of Propranolol for the Treatment of Infantile Cephalic and Facial Hemangiomas : A Clinical Report of 38 Cases / K. Jianguo [et al.] // J Clin Pediatr. Dent. – 2015. – Vol. 39, № 3. – P. 268–271. https://doi.org/10.17796/1053-4628-39.3.268
6. Severe propranolol and ethanol overdose with wide complex tachycardia treated with intravenous lipid emulsion : a case report / J. Jovic-Stosic [et al.] // Clin Toxicol (Phila). – 2011. – Vol. 49, № 5. – P. 426–430. https://doi.org/10.3109/15563650.2011.583251
7. Severe suicidal digoxin and propranolol toxicity with insulin overdose / A. Garg [et al.] // Indian J Crit Care Med. – 2014. – Vol. 18, № 3. – P. 173–175. https://doi.org/10.4103/0972-5229.128709
8. Clarke, E. J. C. Isolation and Identification of Drugs in Pharmaceuticals, Body Fluids and Postmortem Material / E. J. C. Clarke. – London : The Pharm. Press, 2011. – 2463 p.
9. Development and validation of an HPLC-UV method for the simultaneous determination of the antipsychotics clozapine, olanzapine and quetiapine, several beta-blockers and their metabolites / M. Silva Gracia, A. Köppl, S. Unholzer, E. Haen // Biomed Chromatogr. – 2017. – Vol. 31, № 10. https://doi.org/10.1002/bmc.3968
10. Recent Developments in HPLC Analysis of β-Blockers in Biological Samples / K. Saleem, I. Ali, U. Kulsum, H. Y. Aboul-Enein // J. Chromatogr Sci. – 2013. – Vol. 51, № 8. – P. 807–818. https://doi.org/10.1093/chromsci/bmt030
11. Кокорина, Н. О. Определение лекарственных препаратов в биожидкостях методом высокоэффективной жидкостной хроматолграфии / Н. О. Кокорина, В. П. Новоселов, М. А. Ханина // Сибирский медицинский журнал (Томск). – 2008. – Вып.2, № 4. – С. 51–53.
12. Барам, Г. И. Хроматограф “Милихром А–02“. Определение веществ с применением баз данных «ВЭЖХ–УФ» / Г. И. Барам. – Новосибирск : ЗАО Институт хроматографии, 2005. – 64 с.
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