Identification and Quantitative Determination of Clonidine by HPLC Method

Abstract

The aim of this work is identification and quantification of clonidine hydrochloride by a unified HPLC method, which allows to obtain reliable and reproducible research results. Materials and methods. HPLC analysis was carried out on a microcolumn liquid chromatograph "Milichrome A-02" in conditions: reversed-phase variant, column with non-polar sorbent Prontosil 120-5 C 18 AQ, 5 μm; mobile phase in the mode of linear gradient – from eluent А (5 % acetonitrile and 95 % buffer solution) to eluent B (100 % acetonitrile) as during 40 min. The flow rate of the mobile phase has been formed 100 μl/min, injection volume – 4 μl. Multichannel detection of the substance was carried out using a UV detector at 210, 220, 230, 240, 250, 260, 280 and 300 nm; the optimal value of column temperature – 37 - 40°С and pressure of pump – 2.8 – 3.2 MPa. Results and its discussion. As a result of studies using a unified HPLC method, were obtained the retention parameters of clonidine hydrochloride and spectral relationships, which made it possible to include the results obtained in the database for the identification of antihypertensive drugs in the therapeutic monitoring of treatment with an individual drug, or comprehensive treatment of diseases of the cardiovascular system. The development of the quantitative determination of clonidine hydrochloride by HPLC on model solutions using various concentrations of the drug was carried out. The content of clonidine hydrochloride was determined according to the equation S = 0.5 · 10-4 C + 1.8 · 10-3; the correlation coefficient was 0.9964. It is established that the relative uncertainty of the average result did not exceed + 2.12 % when HPLC analysing of clonidine hydrochloride in model solutions. Conclusions. Identification and quantification of clonidine hydrochloride by a unified HPLC method, which allows to obtain reliable and reproducible research results were conducted. The results of research by a unified HPLC method can be recommended for implementation in the practice of forensic bureaus, toxicological centers, clinical laboratories for the study of drugs in biological objects