DEVELOPMENT OF THE HPLC METHOD FOR THE DETERMINATION OF CARBOPLATIN IN MEDICINAL PRODUCTS USING SALTS OF CHAOTROPIC ANIONS

Abstract

Introduction. The leading pharmacopoeias of the world regulate the chromatographic determination of carboplatin using an amino-propyl chromatographic column (class L8). The scientific literature describes the development of HPLC methods for pharmaceuticals based on carboplatin in various matrices using ion-pair reagents, however, such methods have certain drawbacks. In our work, we propose the application of chaotropic anion salts as a promising approach for the development of HPLC methods for carboplatin in pharmaceuticals, which will improve retention and peak shape. The aim of the work was to develop the express, simple, reproducible, and green method for the determination carboplatin in pharmaceuticals using salts of chaotropic anions. Research Methods. For the study, a Shimadzu LC-2050 C liquid chromatograph with DAD was used to obtain chromatograms and integrate results using LabSolutions software. The chromatographic column was a Luna C18 (100 × 4.6 mm, 3 µ m) purchased from Phenomenex. The Carboplatin API (purity ≥ 99 %, HPLC) was procured from Sigma-Aldrich Chemicals Co., and the medicinal product “Ebeve” (10 mg/ml, Austria) was used. Results and Discussion. An important advantage of using salts of chaotropic anions in the mobile phase is the improvement of peak symmetry and retention, as well as the reduction of analysis time. This can be achieved by using low amounts of acetonitrile (ACN) (5–10 %) and high amounts of a buffer solution of 40–50 mM of one of the strongest chaotropes according to Hofmeister’s theory, KPF6 (pH 2.3–2.5) in the mobile phase on C18 or C8 columns. Optimal chromatographic conditions for the determination of carboplatin in a medicinal product have been established: chromatographic column Luna C18 (100 × 4.6 mm, 3 μm), mobile phase – 5 % ACN and 95 % buffer solution KPF6 (40 mM) pH 2.43, column temperature – 30 °C, detection at a wavelength of 195 nm, flow rate – 0.6 ml/min. Linearity was studied in the range of 15–90 μg/ml using the method of least squares, and the regression equation (y = 10148x + 56213) was calculated with a correlation coefficient (R2 = 0.9986). The limit of detection (LOD) was 3.58 μg/ml, and the limit of quantification (LOQ) was 10.86 μg/ml. The most modern tools for studying greenness AGREE (score 0.74), MoGAPI (score 81), Complex MoGAPI (score 81), AGSA (score 77.78), CaFRI (score 82), and CACI (score 79) have established that the proposed HPLC method is ecologically safe. Conclusions. Express, simple, reproducible, and green HPLC method for the determination of carboplatin in pharmaceutical products has been developed using chaotropic anion salts. The proposed HPLC method enables the chromatography and quantitative determination of carboplatin on a C18 octadecylsilane column without the use of ion-pair reagents.