**12. Quantification of mangiferin in the grass of** *Hedysarum caucasicum* **M. Bieb. by HPLC**

High-performance liquid chromatography was used to quantify mangiferin in the subjects under study. Registration of electronic spectra was carried out on a liquid chromatograph Shimadzu Prominence LC-20 AD with a degasser DGU-20A3R. A sample of mangiferin (Sigma-Aldrich, cate. M3547, USA, 2017) was used as a standard sample.

*Standard mangiferin solution*: A slurry of 0.0050 g (based on 100% substance) of mangiferin was placed in a measuring flask with a capacity of 25 ml, dissolved in 70% ethyl alcohol, and then the volume was adjusted with the same solvent to a mark and mixed. An aliquot of 1.0 ml of the resulting solution was placed in a measuring flask with a capacity of 10 ml, dissolved in the mobile phase, and then the volume was adjusted with the same solvent to a mark and stirred. The solution was filtered through a 0.45 μm membrane filter (**Figure 8**).

*Buffer solution*: In a 1000 mL measuring flask, 15.6 g of sodium phosphate of monosubstituted dihydrate was placed, which was dissolved with 200 mL of purified water and the volume adjusted to a mark, stirred. The potentiometric method determined the value of pH, which should be 4.40 0.05.

*Mobile phase*: A mixture of buffer solution, acetonitrile, and methanol in a ratio of 81:16:3.

The developed method of quantitative determination of mangiferin in the above-ground organs of the studied type takes into account the main physicochemical properties of xanthones, and it is characterized by reproducibility, high accuracy, and allows conducting both screening assessment of various raw materials containing mangiferin derivatives and standardization of prepared medicinal plant raw materials of the *Hedysarum* L.

characterized by good reproducibility and tested at raw materials. The obtained results can be used in the preparation of regulatory documentation for plant raw materials. As a result, in species *Hedysarum caucasicum* M. Bieb., *Hedysarum*

**Figure 9.**

**Figure 8.**

*Chromatogram of standard solution of mangiferin.*

Hedysarum *Species from Caucasus*

*DOI: http://dx.doi.org/10.5772/intechopen.91741*

**Figure 10.**

**163**

*Chromatogram of solution 2.*

*Chromatogram of solution 1.*

tive content of tannins in *Hedysarum caucasicum M. Bieb*. was 5.43 0.15%.

*grandiflorum* Pall., and *Hedysarum daghestanicum* Rupr. ex Boiss., biologically active substances such as polysaccharides, triterpene compounds, flavonoids, saponins, polyphenol compounds, leucoanthocyanins, and tannins were found. The quantita-

As a result, it was found that the largest in the above-ground organs of *Hedysarum caucasicum* found was 0.148 0.003% mangiferin (**Figures 9** and **10**). The method of quantification of mangiferin by HPLC has been developed, which is
