DETERMINATION OF MACRO AND MICROELEMENTS CONTENTS IN GENTIANA MACROPHYLLA PALL. HERB

DETERMINATION OF MACRO AND MICROELEMENTS CONTENTS IN GENTIANA MACROPHYLLA PALL. HERB

V. A. Tsukurova ^{* 1}, O. V. Evdokimova ² and V. I. Prokofieva ³

Department of Pharmaceutical and Toxicological Chemistry ³, Institute for Translational Medicine and Biotechnology ¹, Sechenov First Moscow State Medical University, 8-2 Trubetskayast, Moscow, Russian Federation, 119991.

FSBI ² (Scientific Centre for Expert Evaluation of Medicinal Products) of the Ministry of Health of the Russian Federation, Moscow, Russian Federation.

ABSTRACT: Macro and microelements are integrally linked to hormones, vitamins, amino acids, and enzymes in the human body. They affect the normal course of physiological processes. Some medicinal plants contain substantial contents of micro-and macro-elements, which makes it possible to use them purposefully for the prevention and treatment of various diseases. The target of the research was samples of G. macrophylla herb harvested and procured in the Republic of Tyva. The aim of this research provided a quantitative determination of the elemental composition in the herb of G. macrophylla Pall. as a possible source of macro and microelements. The determination was carried out with the atomic absorption spectrophotometry technique. As was discovered, Fe, Mn, Zn, Cu, Cr, Se, Mo, B, and Ag are present in the G. macrophylla herb. The maximum admissible contents of heavy metals in the specimen under study did not exceed the limits recommended for the quality assessment of herbal raw materials.

G. macrophylla Pall., Herbal raw material, Microelements, Macroelements macronutrients, Micronutrients

INTRODUCTION: Macro and microelements play a critical part in human life and activities; they affect enzyme activity, are part of vitamins and hormones, perform the regulatory function, and maintain homeostasis in the body ¹. The following groups are distinguished among macro-and micro-elements: essential (Ca, K, P, Cl, Na, Mg, Zn, Cu, Cr, Fe, I, Mn, etc.), conventionally essential (Ag, Al, Au, B, Li, Si, etc.), conventionally toxic (Al, Bi, Ti, Rb, Sr, Be, etc.) and toxic elements standardized by the State Pharmacopoeia of the Russian Federation (Pb, As, Hg, Cd) ^{2, 3}.

Lack of a certain element in the body may cause various diseases. For example, iron is part of various proteins varied by their function, including enzymes. It takes part in the transport of electrons, oxygen, ensures the course of oxidation-reduction reactions and activation of peroxidation. Under consumption results in hypochromic anemia, myoglobin deficient atony of skeletal muscles, undue fatigability, myocardiopathy, and atrophic gastritis ^{4, 5}.

Manganese participates in the formation of osteal and connective tissue is part of the enzymes participating in the metabolism of amino acids, carbohydrates, and catecholamine is necessary for the synthesis of cholesterol and nucleo-tides. Deficiency of manganese in the body may lead to reproductive system diseases, diabetes, arthronosos, dysimmunity, allergic responses, and arrested development in children ^{6, 7}.

Zinc is part of over 300 enzymes, participates in the processes of synthesis and degradation of carbohydrates, lipos, nucleic acids, and in the regulation of expression of several genes. Under consumption leads to anemia, secondary immunodeficiency, hepatic cirrhosis, sexual dys-function, presence of congenital fetus anomaly ^{8, 9}.

There exist the norms of daily demand and toxicity threshold for essential macro-and microelements for the human as established by the Federal Service for a Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor), which are represented in Table 1.

TABLE 1: NORMS OF DAILY DEMAND IN MACRO AND MICROELEMENTS FOR THE HUMAN²

The following maximum allowable norms of contents (mg/kg) for heavy metals and arsenic in herbal raw material are established: for lead (Pb) – 6.0; arsenic (As) – 0.5; mercury (Hg) – 0.1; cadmium (Cd) – 1.03.

In Tibetan orthodox medicine, the G. macrophylla herb is part of prescriptions to cure the diseases of the respiratory tract, gastrointestinal tract, hepatitis, neurasthenia, influenza, as well as hemostatic teas. In the Chinese orthodox medicine, this herbal raw material is used as a bile-expelling, analgetic, vulnerary, anti-inflammatory, hepato-protective agent for treatment of rheumatism and influenza. In Mongolia, it is used on the fever of epidemic nature as an antifebrile and analgetic agent, in the Russian Federation on freezing injuries and burns, as a blood stopper, vulnerary, immunomodulating agent, and as a digestion amendment agent ^10-12.

The G. macrophylla herb containing flavonoids, iridoids, xanthones, tanning substances, vitamins, organic acids, and other substances are actively used in the orthodox medicine of China, Mongolia, Tibet for a wide spectrum of diseases ^12-14. Data on macro and microelements in the raw material is missing. Previously, a study was conducted on the accumulation of macro and microelements in the roots and flowers of G. macrophylla, collected in different phases of vegetation of the plant ¹⁵. However, the study of the mineral composition of the herb of G. macrophylla was not carried out.

In addition, the content of toxic elements (Pb, As, Hg, Cd) was not determined in this morphological group of raw materials. The purpose of this paper is to determine the elemental composition of G. macrophylla herb as a possible source of macro-, microelements, and the content of toxic elements.

MATERIALS AND METHODS: The target of the research was samples of G. macrophylla herb harvested and procured in the Republic of Tyva in August 2016, in the blooming period. The study of the contents of macro-and microelements of the target object was carried out with the atomic absorption spectrophotometry technique. Deter-mination of micro-and macroelements was carried out on the atomic absorption spectrophotometer AAS KVANT-Z (Russia) equipped with dedicated lamps having a hollow cathode as the radiation source, at a specific wavelength for each element. The Quant Zeeman for Windows software was used to process data. For the purpose of analysis, an assay of the G. macrophylla herb was comminuted to the size of particles that penetrated through a sieve with holes of 1 mm in diameter. About 1.0 g (precisely weighed quantity) of comminuted grass was placed into a porcelain crucible which was then moved to an electric hot plate. It was dried up by gradually increasing the heat and exposed on the plate till the herb started coaling. The crucibles with dried assays were placed into a cold electric furnace. By gradually raising the temperature to 450 °C, mineralization was affected till gray ash has been formed. After liming, the crucibles were cooled to the room temperature, the ash was wetted with water and nitric acid concentrated in (1:1) ratio, evaporated to dryness on the electric hot plate with weak heating, and again the assays were placed into the electric furnace, thus gradually bringing the temperature to 300 °C and и exposing within 30 min. The mineralization was regarded as complete if the ash turned white or slightly stained color without coaled particles.

If any coaled particles remained, the ash was repeatedly processed with water and nitric acid concentrated in (1:1) ratio and then subjected to extra ashing. The produced ash was dissolved while heating in water, and nitric acid concentrated in (1:1) ratio in the amount of 5 ml of acid per weighed quantity. The solution was evaporated to dryness.

The dry residue was dissolved in 20 ml of 1% nitric or hydrochloric acid. On incomplete ash dissolution, the produced nitric acid solution with its residue was evaporated to dryness, and then it was dissolved in 10 ml of hydrochloric acid of 6 M solution and boiled out to wet salts.

The produced salts were dissolved in 20 ml of 1% hydrochloric acid. The solution was filtered through a filter rinsed with a solvent into a 25 ml volumetric flask; the residue on the filter was washed, bringing to the mark using the same solvent and was stirred.

TABLE 2: PARAMETERS FOR DETERMINATION OF MACRO AND MICROELEMENTS WITH THE ATOMIC ABSORPTION SPECTROPHOTOMETRY TECHNIQUE¹⁶

In parallel, 2 references (idle) tests were prepared to determine the purity of chemicals and the glassware by adding all the chemicals into the crucible and precisely reproducing all the conditions (quantity of chemicals, temperature, heating time), in which the assay mineralization was effected, but without the weighed quantity of assay ¹⁶. In parallel, the signal of solutions for standard samples of respective elements of a known concentration was measured Table 2.

The content (X) of an element in the raw materials in mkg/g was calculated with the formula:

X = (Cn –Cx) V. A / a

Where: Сn- Concentration of element in the solution under study calculated with the software program, with the concentration of the standard solution, mkg/ml, taken into account; Сx -concentration of an element in the solution of an “idle” assay calculated using the software program, with the concentration of the standard solution, mkg/ml, taken into account; a - weighed quantity of raw material, g; V - the volume of the solution of assay produced after sample processing, ml; A - dilution factor.

RESULTS: The results of the determination of the mineral composition of G. macrophylla herb are represented in Table 3.

TABLE 3: CONTENTS OF MINERAL SUBSTANCES IN G. MACROPHYLLA HERB

It was shown that G. macrophylla herb contains potassium (K). As was established, the following essential microelements are present in G. macrophylla herb: Fe, Cu, Se, Mn, Mo, (Zn), and Cr, whose concentrations decrease in the following order: Fe>Mn>Zn>Cu>Cr>Se>Mo. Detected was also (B) and silver (Ag) which are related to conventionally essential microelements. The obtained results are indicative of the fact that the contents of cadmium (Cd), mercury (Hg), lead (Pb) and arsenic (As) in G. macrophylla herb does not exceed the allowable levels of values.

CONCLUSION: A research into the determination of the elemental composition of G. macrophylla herb was conducted. As was shown, Fe, Mn, Zn, Cu, Cr, Se, Mo, as well as B and Ag, are present in G. macrophylla herb. This herbal raw material can be a source of the following essential elements: Cr, Mn, Fe, Se. The maximum admissible contents of heavy metals and arsenic in the specimen under study do not exceed the limits recommended for the quality assessment of herbal raw materials.

ACKNOWLEDGEMENT: Nil

CONFLICTS OF INTEREST: The authors declare no conflict of interest.

REFERENCES:

1. Shukla AK, Behera SK, Pakhre A and Chaudhari SK: Micronutrients in Soils, Plants, Animals and Humans. Indian Journal of Fertilisers 2018; 14(4): 30-54.
2. Methodical recommendations of Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor) MR 2.3.1.2432-08. Norms of physiological requirements for energy and nutrients for various population groups of the Russian Federation 2008.
3. Russian Federation State Pharmacopoeia (SPh RF) XIVedition 2018; 2: 2370-85.
4. Fraga CG: Relevance, essentiality and toxicity of trace elements in human health. Molecular Aspects of Medicine 2005; 26: 235-44.
5. Karatela S and Ward NI: Trace elements and human obesity: An overview. Manipal Journal of Nursing and Health Sciences 2016; 2: 50-9.
6. Wood RJ: Manganese and birth outcome. Nutrition Reviews 2009; 67: 416-20.
7. Avila DS, Puntel RL and Aschner M: Manganese in Health and Disease. Metal Ions Life Sciences 2013; 13: 199-27.
8. Chasapis CT, Ntoupa APS, Spiliopoulou CA and Stefanidou ME: Recent aspects of the efects of zinc on human health. Archives of Toxicology 2020; 94: 1443-60.
9. Prasad AS: Discovery of Human Zinc Deficiency: It’s Impact on Human Health and Disease. Advances in Nutrition 2013; 4: 176-90.
10. Jia N, Li Y, Wu Y, Xi M, Hur G, Zhang X, Cui J, Sun W and Wen A: Comparison of the anti-inflammatory and analgesic effects of Gentiana macrophylla and Gentianastraminea Maxim and identification of their active constituents. J of Ethnopharmacology 2012; 144: 638-45.
11. Wu Y, Ai Y, Wang F, Ma W, Bian Q, Lee DY, Dai R: Simultaneous determination of four secoiridoid and iridoid glycosides in rat plasma by ultra performance liquid chromatography tandem mass spectrometry and its application to a comparative pharmacokinetic study. Biomedical Chromatography2015; 30: 97-104.
12. Wang Y, Ahmad B, Duan B, Zeng R and Huang L: Chemical and Genetic Comparative Analysis of Gentiana crassicaulis and Gentiana macrophylla. Chemistry & Biodiversity 2016; 13: 776-81.
13. Hua W, Zheng P, He Y, Cui L, Kong W and Wang Z: An insight into the genes involved in secoiridoid biosynthesis in Gentiana macrophylla by RNA-seq. Molecular Biology Reports 2014; 41: 4817-25.
14. Tsukurova VA, Evdokimova OV, Pavlova LA and Matyushin AA: Large leaf Gentian as a promising source of biologically active substances. Journal of Pharmaceutical Sciences and Research 2017; 9: 2615-17.
15. Niu XX, Chen XW, Su H, Eneji AE, Guo YH and Dong XH: Changes of secondary metabolites and trace elements in Gentiana macrophylla Flowers: A potential medicinal plantpart. Chinese Herbal Medicines 2014; 6(2): 145-51.
16. GOST (State Standard) 31870-2012. Potable Water - Determination of elements content by atomic spectrometry methods (with Corrections). Moscow: Standartinform 2014.
    

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    How to cite this article:

    Tsukurova VA, Evdokimova OV and Prokofieva VI: Determination of macro and microelements contents in Gentiana macrophylla Pall. herb. Int J Pharm Sci & Res 2021; 12(3): 1442-45. doi: 10.13040/IJPSR.0975-8232.12(3).1442-45.

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