**13. Amino acid analysis**

Amino acid analysis in the grass of species of the genus *Hedysarum* was carried out on the basis of the FSBOU VO of the Stavropol GAU by column liquid chromatography on an amino acid analyzer according to the procedure indicated in the guest 32,195–2013. Qualitative and quantitative analysis of amino acids was carried out on an amino acid analyzer, AminoAcid Analyzer AAA 339 M (Mikrotechna). The comparative amino acid composition of the three studied samples of species of the genus growing in the North Caucasus showed that amino acids such as aspartic (1.84–2.68%) and glutamic acid (1.29–1.33%) as well as proline (0.83–1.39%), leucine (0.86–0.90%), and phenylalanine (0.58–0.63%) were found in significant quantities in the above-ground organs of the studied species. **Table 7** shows the results of a comparative analysis of the amino acid composition of the species under study.

The study of the amino acid composition during complex pharmacognostic analysis is one of the mandatory areas of research, since amino acids are involved in the biogenesis of many biologically active compounds, including xanthones. It is known that the main components of xanthone biosynthesis are acetyl-CoA,

mevalonic, and shikimic acids, from which phenylalanine is further synthesized. The obtained research results can be further used in preparing a complex metabolomic evaluation of medicinal plant raw materials of *Hedysarum* species. Spectral analysis of the above-ground organs of *Hedysarum caucasicum* M. Bieb. was carried out at the Central Testing Laboratory of Caucasian Geologic Survey JSC in Essentuki. Sample preparation of grass raw material *Hedysarum caucasicum* M. Bieb. was carried out according to the methodology of the Pharmacopea of Russian Federation. Ash was studied according to the method of the MP plant 4С atomic

**Element title Raw material content, % Element title Raw material content, %**

K 1.47 Na 0.10 Ca 0.29 P 0.49

Al 0.049 Mn 0.0025 Ba 0.0029 Cu 0.00098 B 0.00098 Ti 0.0073 Fe 0.029 Cr 0.0009 Si 0.29 Zn 0.0049

The ash sample was evaporated into the graphite electrode cell by means of an electric arc. In equilibrium processes of excitation and reverse transition to the basic state of electrons of element atoms, emission (emission) spectra were recorded. The

For the first time, morphological-anatomical diagnostic signs of the species are necessary for standardization of medicinal vegetal raw materials. Indicators of caulifolar micromorphology were introduced to diagnose plant objects using the example of the genus *Hedysarum* L. Molecular genetic studies were carried out and morphometric indicators were determined, which make it possible to establish correlations between morphological, molecular genetic, and phytochemical indicators of species assigned to certain sections of the genus, as well as to predict the

Introduction studies of *Hedysarum causicum M. Bieb.*, *Hedysarum grandiflorum Pall.*, and *Hedysarum daghestanicum Rupr. ex Boiss.* on the territory of the Botanical Garden of the PMFI and in the Dagestan Scientific Center of the Mining Botanical Garden of the Russian Academy of Sciences, including the main phases of develop-

Spectral analysis of herb *Hedysarum caucasicum* M. Bieb. represented by macroelements (potassium, calcium, magnesium, sodium, phosphorus) and trace elements (manganese, iron, zinc, copper, and silicon). The obtained results of elemental composition are necessary in the complex analysis of the sum of active

emission spectrometry on diffraction spectrograph.

*Elemental composition of herb* Hedysarum caucasicum *M. Bieb.*

accumulation of xanthones in previously unearthed species.

ment, phenological spectra, were compiled.

results are shown in **Table 8**.

substances of the plant.

**14. Conclusion**

**165**

Macroelements

Microelements

**Table 8.**

Mg 0.10

Hedysarum *Species from Caucasus*

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


*daghestanicum Rupr. ex Boiss.*

#### **Table 7.**

*Comparative analysis of amino acid composition in three samples of Hedysarum L. species growing in the North Caucasus, %.*

#### Hedysarum *Species from Caucasus DOI: http://dx.doi.org/10.5772/intechopen.91741*


#### **Table 8.**

**13. Amino acid analysis**

*Legume Crops - Prospects, Production and Uses*

Monoamino dicarboxylic acids

Monoamine monocarboxylic acids

Monoamine monocarboxylic acids

Heterocyclic compounds

*daghestanicum Rupr. ex Boiss.*

**Table 7.**

**164**

*Caucasus, %.*

study.

Amino acid analysis in the grass of species of the genus *Hedysarum* was carried out on the basis of the FSBOU VO of the Stavropol GAU by column liquid chromatography on an amino acid analyzer according to the procedure indicated in the guest 32,195–2013. Qualitative and quantitative analysis of amino acids was carried out on an amino acid analyzer, AminoAcid Analyzer AAA 339 M (Mikrotechna). The comparative amino acid composition of the three studied samples of species of the genus growing in the North Caucasus showed that amino acids such as aspartic (1.84–2.68%) and glutamic acid (1.29–1.33%) as well as proline (0.83–1.39%), leucine (0.86–0.90%), and phenylalanine (0.58–0.63%) were found in significant quantities in the above-ground organs of the studied species. **Table 7** shows the results of a comparative analysis of the amino acid composition of the species under

The study of the amino acid composition during complex pharmacognostic analysis is one of the mandatory areas of research, since amino acids are involved in the biogenesis of many biologically active compounds, including xanthones. It is known that the main components of xanthone biosynthesis are acetyl-CoA,

**Types Structure №1 №2 №3**

Asparaginic *α*-Aminosuccinic acid 1.86 2.68 1.84 Glutaminic *α*-Aminoglyutaroic acid 1.33 1.31 1.29

Leucine\* *α*-Aminoisocapronic acid 0.90 0.89 0.86 Valine\* *α*-Aminoisovaleric acid 0.68 0.74 0.71 Alanine *α*-Aminopropionic acid 0.67 0.65 0.63 Serine *α*-Amino-*β*oxypropionic acid 0.62 0.66 0.60 Phenylalanine\* *α*-Amino-*β*-phenylpropionic acid 0.62 0.63 0.58 Glycine *α*-Aminoacetic acid 0.55 0.58 0.57 Threonine\* *α*-Amino-*β*-hydroxy-butyric acid 0.54 0.51 0.50 Thyrosine *α*-Amino-*β*-oxy phenylpropionic acid 0.52 0.63 0.54 Isoleucine\* *α*-Amino-*β*-ethyl-*β*-methylphenylpropionic acid 0.48 0.58 0.54 Methionine\* *α*-Amino-*γ*-methylthion-n-butyric acid 0.04 0.11 0.10

Lysine\* *α*, *ε*-Aminocapronic acid 0.74 0.73 0.72 Arginine *α*-Amino-*σ*-guanidine-n-valeric acid 0.55 0.56 0.56

Proline Pyrrolidine-*α*-carboxylic acid 0.83 1.39 1.19 Gystidine *α*-Amino-*β*-imidazolyl-propionic acid 0.33 0.42 0.35

*Comparative analysis of amino acid composition in three samples of Hedysarum L. species growing in the North*

*\*№1—Hedysarum caucasicum M. Bieb.; №2—Hedysarum grandiflorum Pall.; and №3—Hedysarum*

**Amino acid type Species**

*Elemental composition of herb* Hedysarum caucasicum *M. Bieb.*

mevalonic, and shikimic acids, from which phenylalanine is further synthesized. The obtained research results can be further used in preparing a complex metabolomic evaluation of medicinal plant raw materials of *Hedysarum* species.

Spectral analysis of the above-ground organs of *Hedysarum caucasicum* M. Bieb. was carried out at the Central Testing Laboratory of Caucasian Geologic Survey JSC in Essentuki. Sample preparation of grass raw material *Hedysarum caucasicum* M. Bieb. was carried out according to the methodology of the Pharmacopea of Russian Federation. Ash was studied according to the method of the MP plant 4С atomic emission spectrometry on diffraction spectrograph.

The ash sample was evaporated into the graphite electrode cell by means of an electric arc. In equilibrium processes of excitation and reverse transition to the basic state of electrons of element atoms, emission (emission) spectra were recorded. The results are shown in **Table 8**.

Spectral analysis of herb *Hedysarum caucasicum* M. Bieb. represented by macroelements (potassium, calcium, magnesium, sodium, phosphorus) and trace elements (manganese, iron, zinc, copper, and silicon). The obtained results of elemental composition are necessary in the complex analysis of the sum of active substances of the plant.

## **14. Conclusion**

For the first time, morphological-anatomical diagnostic signs of the species are necessary for standardization of medicinal vegetal raw materials. Indicators of caulifolar micromorphology were introduced to diagnose plant objects using the example of the genus *Hedysarum* L. Molecular genetic studies were carried out and morphometric indicators were determined, which make it possible to establish correlations between morphological, molecular genetic, and phytochemical indicators of species assigned to certain sections of the genus, as well as to predict the accumulation of xanthones in previously unearthed species.

Introduction studies of *Hedysarum causicum M. Bieb.*, *Hedysarum grandiflorum Pall.*, and *Hedysarum daghestanicum Rupr. ex Boiss.* on the territory of the Botanical Garden of the PMFI and in the Dagestan Scientific Center of the Mining Botanical Garden of the Russian Academy of Sciences, including the main phases of development, phenological spectra, were compiled.

Within the framework of phytochemical screening, BAVs such as polysaccharides, triterpene compounds, flavonoids, saponins, polyphenolic compounds, leucoanthocyanins, and tannins were found in the objects we studied. For the first time in the grass of three species of the genus *Hedysarum*, the presence of amino acids, the main part of which belongs to the group of essential amino acids, and the presence of proline and phenylalanine prove the presence of xanthones. The obtained research results can be further used in preparing a complex metabolomic evaluation of medicinal plant raw materials of *Hedysarum* species.

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A comprehensive study of the qualitative analysis and quantitative content of the sum of xanthones and, in fact, mangiferin was carried out using thin-layer and paper chromatography, UV spectrophotometry, capillary zone electrophoresis, as well as high-performance liquid chromatography.

In the framework of complex pharmacognostic studies of three species of the genus *Hedysarum*, we have developed a method for quantitative determination by UV spectrophotometry of the sum of xanthones in terms of mangiferin. The technique takes into account the basic physicochemical properties of xanthones; is characterized by the reproducibility, high accuracy, and simplicity of execution; and allows conducting both a screening assessment of various raw materials containing mangiferin derivatives and standardization of the prepared vegetable raw materials.

The developed methods are tested on the above-ground organs of *Hedysarum* species, collected and dried taking into account the rules and requirements for the preparation of medicinal raw materials. As a result, it was found that the greatest quantitative content of the sum of xanthones in terms of mangiferin is distinguished by the grass *Hedysarum caucasicum* M. Bieb. (0.62 0.021%). The results show the prospect of further investigation of *Hedysarum caucasicum* M. Bieb. herb as an additional source of mangiferin.

#### **Conflict of interest**

The authors declare no conflicts of interest.

#### **Author details**

Serebryanaya Fatima<sup>1</sup> \* and Imachueva Djavgarat<sup>2</sup>

1 Department of Pharmacognosy, Botany and Technology of Phytopreparations of Pyatigorsk Medical Pharmaceutical Institute, Pyatigorsk, Russia

2 Daghestan State Medical University of the Ministry of Health of Russia, Mahachkala, Daghestan Republic, Russian Federation

\*Address all correspondence to: f.k.serebryanaya@pmedpharm.ru

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Hedysarum *Species from Caucasus DOI: http://dx.doi.org/10.5772/intechopen.91741*
