Validation evaluation of the method of quantitative determination of α(1,2)-L-rhamno-α(1,4)-D-galactopyranosyluronane Acorus calamus L. in dosage form. Problems of biological, medical and pharmaceutical chemistry

Introduction. The object of increased attention of researchers is calamus (Acorus calamus L.), for the components of which a wide range of pharmacological activities have been identified – immunomodulatory, anti-inflammatory, analgesic, antioxidant, neuroprotective, hypolipidemic, antitumor. According to previous studies, it was found that α(1,2)-L-rhamno-α(1,4)-D-galactopyranosyluronan, isolated from the rhizomes of Acorus calamus L., has antimetastatic, antiblastoma, and immunomodulatory properties. The substance can also reduce the toxic effect on the hematopoietic system and increase the effectiveness of chemotherapy. Based on it, a finished dosage form was obtained - a solution for intravenous administration of 10 mg/ml, which requires the development of valid quality control methods for inclusion in the draft regulatory document on the quality and registration of a new drug. The aim of the study was to determine the metrological characteristics of the chromatographic technique for the quantitative determination of α(1,2)-L-rhamno-α(1,4)-D-galactopyranosyluronan in the finished dosage form.

Material and methods. The object of the study was the finished dosage form - a solution for intravenous administration of α(1,2)-L-rhamno-α(1,4)-D-galactopyranosyluronan Acorus calamus L. 10 mg/ml, produced on the basis of the educational establishment of the Technology Implementation Center of the Federal State Budgetary Educational Institution of Higher Education of the Siberian State Medical University of the Ministry of Health Russia. Quantitative determination was carried out using a Dionex Ultimate 3000 liquid chromatograph (Thermo, Germany) with a refractometric detector RI-101. Validation assessment was carried out according to the following indicators: linearity, accuracy, precision under repeatability conditions.

Results. Linear regression analysis of the obtained results using the least squares method showed that the technique is linear in the concentration range of 80–120%. When determining the accuracy, the openability for concentrations of 80, 100 and 120% varied in the range of 99.68–101.96%. The values of the relative standard deviation did not exceed the permissible values when assessing precision 0.82–4.51%.

Conclusions. The methodology is valid for the studied indicators and can be recommended for inclusion in the draft regulatory document on quality.