The investigations of propranolol by HPLC method suitable for a chemical-toxicological analysis

Abstract

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.