Validation of the methodology for quantitative determination of flavonoids in an immunomodulatory collection

Introduction. Currently, the number of diseases associated with disorders in the immune system is increasing worldwide. In this regard, it is important to study and develop new drugs for the prevention and maintenance of the immune system, which have a mild and long-lasting effect, minimal side effects and no contraindications, and are also available to the general population. These requirements are met by herbal preparations containing various groups of biologically active substances (BAS), in particular flavonoids. The immunomodulatory effect of flavonoids is carried out through a number of mechanisms, including the regulation of pro- and anti-inflammatory cytokines. In order to develop new herbal drugs and their subsequent approval for medical use, it is necessary to create a regulatory document regulating their quality. One of the key quality indicators is the "Quantitative Determination" section, which is a method for quantitatively determining the content of the amount of BAS, which must be validated.

The aim of the work – validation of the methodology for quantitative determination of flavonoids in a collection with immunomodulatory activity.

Material and methods. The object of the study was a collection consisting of Rosehips (Rosae fructus), Hawthorn fruits (Crataegi fructus), Licorice roots (Glycyrrhizae radices), Common marigold flowers (Calendula officinalis flores), Purple coneflower herb (Echinaceae purpurea herba). Quantitative determination of the sum of flavonoids was carried out by spectrophotometric method on spectrophotometer (Shimadzu UV-1800, Japan) after complexation reaction with alcohol solution of aluminum chloride. Validation was carried out in accordance with OFS.1.1.0012 of the State Pharmacopoeia of the XV edition. Statistical processing of results was carried out using Microsoft Excel software package.

Results. The studies of the quantitative content of the amount of flavonoids in the immunomodulatory collection, consisting of five types of medicinal plant raw materials approved for use in medicine, were carried out by differential spectrophotometry. A validation evaluation of the methodology for quantifying the amount of flavonoids in terms of rutin was carried out. The analysis of the results showed that the parameters correctness, linearity, convergence, intra–laboratory and interlaboratory precision were within the criteria of acceptability; the error of determination did not exceed 5%, the percentage of recovery was within 99.31–100.87%, which corresponds to the criterion of correctness (100 ± 5.00%).

Conclusions. The conducted studies of the validation evaluation of the methodology for quantifying the amount of flavonoids in terms of rutin in the immunomodulatory collection make it possible to include the developed methodology in the draft regulatory documentation in the "Quantification" section.