Development of methods of identification and quantitative determination of active substances in semi-solid dosage forms with sapropel extracts

Abstract

For use in veterinary medicine for the treatment of the wound process and the prevention of mastitis, the composition and technology of an emulsion-based cream, conventionally called “Saprocream”, has been developed. Clinical trial of the drug “Saprocream” proved its effectiveness and safety for use in the healing of microtraumas, erosions and cracks (wounds) of the udders of cows. The aim of the research were was the standardization of the cream under the conventional name “Saprocream” for use in veterinary medicine as a wound healing agent, the development of methods of identification and quantification and their validation. Materials and methods. Test samples of emulsion cream type o / v, containing 15 % aqueous extract of sapropel (VES), 10 % oil extract of sapropel (OES), 6 % emulsifier No. 1, 1 % cetylstearyl alcohol, preservatives – 0.01 % nisin, 0.1 % euxil K 100 and purified water up to 100 g were made by phase inversion. The methods of pharmacopoeial articles of the European Pharmacopoeia of the 10th edition, SPhU 1.0, as well as industry standards were used to develop methods for identification and quantification of active pharmaceutical ingredients (API) in the developed cream and its standardization. Results. The chromatographic profile of the acetone extract from the cream coincides with the chromatographic profile of the reference solution ((PRS) β-carotene), which indicates the presence of substances of carotenoid structure. The absorption spectrum of hexane extraction from the cream in the range from 500 nm to 750 nm has a maximum absorption at a wavelength of 670±2 nm, which coincides with the maximum absorption of hexane solution OES, indicating the presence of chlorophyll. A method for quantifying the total mass fraction of humic acids (HA) has been developed and it has been established that the total mass fraction of HA in cream samples with sapropel extracts is 0.828 %. The results of the validation of the method show that there is a linear relationship between the concentration of the total mass fraction of HA and the mass of the AES sample with a correlation coefficient of 0.9981 (³ 0.9981). The developed technique is precise, because the value of the relative confidence interval is less than the critical value for the convergence of the results: D %=1.34≤1.60 and the criterion of insignificance of systematic error d=0.51 is fulfilled. Conclusions. An emulsion-based cream under the conventional name “Saprocream” has been standardized, methods for identification and quantification of active substances have been developed and validation of the developed methods for use in veterinary medicine as a wound-healing and anti-inflammatory agent has been carried out.