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| ORIGINAL ARTICLE |
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| Year : 2019 | Volume
: 10
| Issue : 2 | Page : 56-62 |
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Study and modeling of the distribution process of some phenolic compounds between the solid and liquid phases
Nikolay Boyko1, Dmitriy Pisarev1, Elena Zhilyakova1, Alina Pravlotskaya1, Oleg Novikov2, Nikolay Makarevich3, Viktoria Kuznietsova4, Natalia Sushchuk5
1 Department of Pharmaceutical Technology, Belgorod State University, Belgorod, Russia
2 Test Department “Drugs Quality Control Center” of the Shared Research and Education Center, Peoples' Friendship University of Russia (RUDN University), Moscow, Russia
3 Department of Pulp and Paper and Wood Chemical Industries, Higher School of Natural Sciences and Technologies, M.V. Lomonosov Northern (Arctic) Federal University, Arkhangelsk, Russia
4 Department of Chemistry of Natural Compounds, National University of Pharmacy, Kharkiv, Ukraine
5 Department of Technology of Drugs, Odessa National Medical University, Odessa, Ukraine
| Date of Web Publication | 12-Apr-2019 |
Correspondence Address:
Prof. Nikolay Boyko
Department of Pharmaceutical Technology, Belgorod State University, Belgorod
Russia
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/japtr.JAPTR_392_18
The article presents the results related to the study of distribution of biologically active substances from the plant raw material between solid and liquid phases. The aim of this study is to develop theoretical bases of the extraction process in the equilibrium state by the example of study and modeling of the distribution process of biologically active substances from Eucalyptus viminalis leaves. In these studies, we used ground plant raw material of E. viminalis leaves with particle fraction of 0.1–0.5 mm; and ethanol with concentration 80% ±1% v/v was used as an extractant. Qualitative and quantitative analyses were carried out by reversed phase high-performance liquid chromatography with rutin, chlorogenic acid, and euglobal standards equivalent to spissum extract of chlorophyllipt of the State Pharmacopoeia of Ukraine. A hypothesis has been suggested that Henry's adsorption law and the law of conservation of matter play a fundamental role in this process. The experimental data are described well by the suggested equation with high value of determination coefficient R2 =0.99. At the same time, F-test and the significance of coefficients in equations satisfy the statistic condition, which means that the current hypothesis about the adsorption mechanism of distribution of biologically active substances in the extraction system is not refuted. The results of these studies demonstrate good agreement of experimental data and theoretical model based on Henry's adsorption law and mass balance. The numerical values of constants in the model suggested have been calculated.
Keywords: Distribution, equilibrium, Eucalyptus viminalis Labill, leaves, phenolic compounds
How to cite this article:
Boyko N, Pisarev D, Zhilyakova E, Pravlotskaya A, Novikov O, Makarevich N, Kuznietsova V, Sushchuk N. Study and modeling of the distribution process of some phenolic compounds between the solid and liquid phases. J Adv Pharm Technol Res 2019;10:56-62 |
How to cite this URL:
Boyko N, Pisarev D, Zhilyakova E, Pravlotskaya A, Novikov O, Makarevich N, Kuznietsova V, Sushchuk N. Study and modeling of the distribution process of some phenolic compounds between the solid and liquid phases. J Adv Pharm Technol Res [serial online] 2019 [cited 2023 Apr 1];10:56-62. Available from: https://www.japtr.org/text.asp?2019/10/2/56/256019 |
| Introduction | |  |
The development of drugs including phytodrugs is still on top of its relevancy.[1],[2],[3] At present, among the drugs urgently needed, there are those having antimicrobial activity due to widespread of microorganisms with antibiotic resistance.[4] Herewith, active studies are carried out all over the world to solve this problem. One of the possible ways to solve this problem is combined use of antibiotics with other substances that improve their activity.[5]
A good alternative to drugs based on the synthetic substances or antibiotics used for local treatment is the use of phytodrugs that have additional useful properties, not only antimicrobial activity. In our previous works, we studied antimicrobial activity of extracts from plant raw materials and tinctures that contained phenolic compounds, for example, Amorpha fruticosa L.(fruits), Centaurium erythraea (herb), Paeonia anomala L. (root), Cetraria islandica (thallus), Dryopteris filix-mas L. (leaves and root), Humulus lupulus L. (cones), Salix alba L. (cortex), Pentaphylloides fruticosa L. (herb), and Eucalyptus tincture.[6],[7],[8] These results demonstrate that extracts from H. lupulus L., D. filix-mas L., and Eucalyptus viminalis L. on ethanol 70% v/v have a significant level of antimicrobial activity, which is confirmed by data from scientific literature.[9],[10],[11]
According to literary sources, phloroglucinol derivatives (flavaspidic acids, xanthohumol, and euglobals) are responsible for antimicrobial activity; moreover, these compounds have some other important activities, such as anthelmintic, antiviral, antitumor, anti-inflammatory, and antioxidant.[12],[13],[14],[15],[16],[17]
Thus, plants that contain this group of biologically active substances are very promising for further study and development or improvement of phytodrugs technology, especially those having antibacterial activity. Plants of Eucalyptus genus are widely used in medicine all over the world.
The plants of Eucalyptus genus, Myrtaceae family, have been used for medicinal purposes for centuries due to their numerous useful activities (antiseptic, expectorant, anti-inflammatory, insecticide, repellent, etc.).[18],[19] According to scientific sources, eucalyptus leaves contain different groups of substances: essential oil up to 6% (cineole), tannins up to 11%, triterpene saponins 2%–4% (ursolic acid derivatives), phenol carbonic acids (gallic, chlorogenic), flavonoids (rutin, hyperoside, and eucaliptine), and euglobals (acyl-phloroglucinol-monoterpenes and acyl-phloroglucinol-sesquiterpenes/macrocarpale).[20] At the same time, euglobals are a very important group of substances due to their antileishmanial, antiviral, and antimicrobial activities.[21],[22] Therefore, the studies in the field of extraction of biologically active substances that have antimicrobial activities from plant raw material of Eucalyptus genus are an urgent task.
The stage of extraction of biologically active substances from the plant raw material is a necessary part in the technology of any phytodrug. A future technology of the drug and eventually its quality in many instances depend on how this process is organized. One of the important technological parameters for the extraction system is the equilibrium concentration of biologically active substances in the extractant under conditions of its dynamic equilibrium state onset. This parameter determines the expulsive force of mass exchange process of biologically active substances from the plant raw material particles into a free extractant and therefore determines the velocity of the extraction process. Moreover, this parameter determines the extract's therapeutic value and the number of drug's units per production lot and consequently the profit.
Therefore, forecast of this parameter is among important tasks in phytotechnology. We have not found any information in scientific sources on a possible distribution mechanism of biologically active substances from this plant raw material in the extraction system between the solid and liquid phases and, therefore, development of a mathematical model that describes this process seems to be an actual task.
Thus, the aim of this study is to develop theoretical bases of the extraction process in the equilibrium state by the example of study and modeling of the distribution process of biologically active substances from E. viminalis leaves.
| Materials and Methods | | |
Plant raw material
For the study, we used pharmacopoeia plant raw material E. viminalis leaves from “Krasnogorskleksredstva” company, Krasnogorsk, Russia, batch No. 100917, best before 10/2020.
For extraction, we used the ground plant raw material with particle fraction between 0.1 and 0.5 mm, and we used ethanol with concentration 80% ±1% v/v as an extractant.
Chemicals and reagents
Qualitative and quantitative analyses of dominative biologically active substances were carried out by reversed phase high-performance liquid chromatography (RP HPLC) with rutin and chlorogenic acid standards; due to the fact that euglobal standard is not easily available, we carried out quantitative calculations in equivalent to spissum extract of chlorophyllipt of the State Pharmacopoeia of Ukraine. We used ethanol of pharmaceutical grade, manufactured in Russia.
Method of extracts obtaining
The equilibrium process in the extraction system was studied at 4, 20, 40, and 60°C ± 1°C; and a method of simple maceration for 24 h was used. Distribution of biologically active substances between the phases was studied at weight of plant raw material/volume of the extractant ratio of 1:5, 1:10, 1:20, and 1:40.
High-performance liquid chromatography analysis
RP HPLC analysis was carried out using a chromatograph by “Agilent Technologies,” “Agilent 1200 Infinity” series, made in the USA. RP HPLC analysis was carried out under the following conditions: 1% water solution of formic acid was used as mobile phase (A); ethanol 96% v/v was used as second mobile phase (B); mobile phases were pumped in a linear gradient elution regime; chromatographic column was Supelco Ascentis express C18 100 mm × 4.6 mm with particle size of 2.7 μm; the velocity of the mobile phase was 0.5 ml/min; the temperature of chromatographic column was + 35°C; and the sample volume was 1 μl. A detailed description of chromatography conditions is presented in the article.[23]
RP HPLC analysis was carried out using a diode-array detector at the following wavelengths: 325 nm for chlorogenic acid, 350 nm for rutin, and 275 nm for euglobal.
Suitability and validation parameters for method of analysis
The main validation parameters of the analytical method and suitability of the HPLC system for determination of rutin, chlorogenic acid, and euglobal equivalent to spissum extract of chlorophyllipt are presented in [Table 1].[24] | Table 1: Main validation parameters of the analytical method and suitability of the HPLC system for determination of rutin, chlorogenic acid, and euglobal equivalent to spissum extract of Chlorophyllipt
Click here to view |
Theoretical part
To explain and determine the possibility of mathematical modeling of the distribution process of biologically active substance molecules in the extraction system, we hypothesized that Henry's adsorption law and the law of conservation of matter should make the background of this process.
Under this assumption, distribution of biologically active substances between the phases should have linear dependency of the reverse value of biologically active substances concentration in the extract from the volume of the extractant at constant temperature (1):
where C is biologically active substance concentration, g/ml; m0 is total quantity of biologically active substances in the extraction system, g; M is solid unsolved part of the plant raw material, g; V is volume of the extract, to simplify, we take it as volume of the extractant in the extraction system, ml; KH is Henry's constant, which is  , ml/g; ΔG is energy constant of the distribution process of biologically active substances, J/mole; R is gas constant, which is equal 8.314 J/(mole·K); T is absolute temperature, K; φ is dimensionless constant; a is a constant equal to reverse value of total weight of biologically active substances in the extraction system, 1/g; b is constant, which is (M φ/m0)·exp [ΔG/(RT)], ml/g.
The value of constants (ΔG) and (M·φ) can be calculated using regression equation (2):
Data analysis
Regression analysis of data was carried out in MS Office Excel 2010 with data analysis tool. The results were obtained at repeat count n = 3 and confidence coefficient P = 0.95.
| Results | | |
[Figure 1] presents a typical chromatogram of the extract (plant raw material/extractant ratio 1:10, at 20°C ±1°C) obtained by RP HPLC analysis using a diode-array detector at wavelength of 350 nm (for rutin). | Figure 1: Reversed-phase high-performance liquid chromatography chromatogram of the extract at 350 nm. I is rutin with ultraviolet spectra
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[Figure 2] presents a typical chromatogram of the extract (plant raw material/extractant ratio 1:10, at 20°C ±1°C) obtained by RP HPLC analysis at wavelength of 325 nm (for chlorogenic acid). | Figure 2: Reversed-phase high-performance liquid chromatography chromatogram of the extract at 325 nm. II is chlorogenic acid with ultraviolet spectra
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[Figure 3] presents a typical chromatogram of extract (plant raw material/extractant ratio 1:10, at 20°C ±1°C) obtained by RP HPLC analysis at wavelength of 275 nm (for euglobals). | Figure 3: Reversed-phase high-performance liquid chromatography chromatogram of the extract at 275 nm for euglobals. III is the dominant euglobal with ultraviolet spectra
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As it can be seen from chromatograms in [Figure 1], [Figure 2], [Figure 3], in the extract on the basis of ethanol-water solution 80% v/v, we detected euglobals by RP HPLC analysis (at 275 nm, retention time from 40 to 51 min), which are dominant compared to all other compounds detected, as well as rutin (at 350 nm, retention time: Minute 17) and chlorogenic acid (at 325 nm, retention time: Minute 7), which agrees well with other sources mentioned above.
[Figure 4] presents the results after experimental data processing in coordinates 1/C = f (V) for rutin at different temperature values and plant raw material/extractant ratios. | Figure 4: Experimental data and regression equations for rutin at different temperature values
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[Figure 5] presents the results after experimental data processing in coordinates 1/C = f (V) for chlorogenic acid at different temperature values and plant raw material/extractant ratios. | Figure 5: Experimental data and regression equations for chlorogenic acid at different temperature values
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[Figure 6] presents the results after experimental data processing in coordinates 1/C = f (V) for euglobal equivalent to spissum extract of chlorophyllipt at different temperature values and plant raw material/extractant ratios. | Figure 6: Experimental data and regression equations for euglobal equivalent to spissum extract of chlorophyllipt at different temperature values
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As it can be seen from [Figure 3], [Figure 4], [Figure 5], [Figure 6], experimental data are described well by equation (1) and have linear dependency with a high value of determination coefficient that is equal R2 =0.99.
In addition, F-test satisfies the condition Fcalc≤ Ftable, and the significance of coefficients in regression equations satisfies the condition pcalc≤ ptable, which confirms the adequacy of regression equations. Therefore, the theoretical model suggested is probably significant.
Subsequently, we calculated Henry's constant (KH=b/a), energy constant (ΔG), and dimensionless constant (M·φ) using the regression equation constants obtained at different temperature values.
[Figure 7] shows the regression equations between the logarithm of Henry's constant and reverse value of temperature for rutin, chlorogenic acid, and euglobal equivalent to spissum extract of chlorophyllipt. | Figure 7: The dependency between the logarithm of Henry's constant and reverse value of temperature for rutin, chlorogenic acid, and euglobal equivalent to spissum extract of chlorophyllipt
Click here to view |
As it can be seen from the data in [Figure 7], experimental data are described well by equation (2) and have linear dependency with a high value of determination coefficient R2 =0.99.
At the same time, F-test satisfies the condition Fcalc≤ Ftable, and the significance of coefficients in regression equations satisfies the condition pcalc≤ ptable, which confirms the adequacy of regression equations. Therefore, the theoretical model suggested is probably significant, and the suggested hypothesis about the distribution mechanism of biologically active substances in the extraction system is not refuted.
[Table 2] presents the values of constants that were calculated: total quantity of biologically active substances in the extraction system (m0) by equation (1); energy constant (ΔG) and dimensionless constant (M·φ) by equation (2). | Table 2: Values of theoretical constants for biologically active substances
Click here to view |
As it can be seen from [Table 2], energy parameters (ΔG) for rutin, chlorogenic acid, and euglobal equivalent to spissum extract of chlorophyllipt have the same value; in general, it equals 23.3 kJ/mole, which is typical for energy of the process of physical adsorption.
The value of dimensionless constant (M·φ) for rutin, chlorogenic acid, and euglobal equivalent to spissum extract of chlorophyllipt is equal to 0.000050, 0.000025, and 0.000091, respectively. When making a comparison of these values with those of total quantity of biologically active substances in the extraction system (m0), it can be observed that they are <1% of the total quantity of the substance in the raw material, and probably, it is the limit value of the substance absorbed by the solid phase of the plant raw material.
Moreover, it is interesting to compare the values of total quantity of biologically active substances in the extraction system (m0) calculated with their respective experimental values. For this purpose, [Table 3] shows the main parameters of plant raw material.
As it can be seen from data in [Table 2] and [Table 3], the calculated value of total quantity of biologically active substances in the extraction system (m0) coincides with its experimental value within inaccuracy range for rutin (820 ± 170)·10−5 ≈ (860 ± 40)·10−5, for chlorogenic acid (190 ± 30)·10−5 ≈ (200 ± 10)·10−5, and for euglobal equivalent to spissum extract of chlorophyllipt (10700 ± 2600)·10−5 ≈ (12400 ± 1100)·10−5 g/g plant raw material.
| Discussion | | |
The results obtained in these studies are in good agreement with our previous work, where we used it for modeling of glycyram and licurosid distribution between Glycyrrhizae radix and 70% v/v ethanol.[25]
Thus, based on the mentioned above, we have concluded that experimental data are described well by the mathematical model suggested and the hypothesis about the adsorption mechanism of biologically active substances distribution in the extraction system from E. viminalis leaves and 80% v/v ethanol is not refuted.
The hypothesis and mathematical model suggested allow explaining and forecasting the distribution of biologically active substances between the phases under the equilibrium state.
Our next step will be using this model to forecast the concentration of biologically active substances for the method of fractional maceration and even for nonequilibrium filtration extraction method.
In the case of correspondence of experimental data and the model suggested, it will be possible to explain one of the two main sides of the extraction process of biologically active substances from the plant raw material, namely, the equilibrium state in the extraction system.
| Conclusion | | |
Distribution of rutin, chlorogenic acid, and euglobal equivalent to spissum extract of chlorophyllipt between the phases of the extraction system from E. viminalis leaves in ethanol with concentration 80% v/v has been studied.
A hypothesis about the adsorption mechanism of distribution of biologically active substances in the extraction system has been suggested and used for the development of a mathematical model to describe the experimental data obtained.
The results of our studies demonstrate good agreement of experimental data and mathematical model based on Henry's low and mass balance.
Financial support and sponsorship
The results were obtained under state grant No. 12.6429.2017/BCh “Complex research of plant-origin objects in the process of creating targeted dosage forms for proctology”.
Conflicts of interest
There are no conflicts of interest.
| References | | |
| 1. | Cragg GM, Newman DJ. Natural products: A continuing source of novel drug leads. Biochim Biophys Acta 2013;1830:3670-95.  |
| 2. | Newman DJ, Cragg GM. Natural products as sources of new drugs over the 30 years from 1981 to 2010. J Nat Prod 2012;75:311-35. |
| 3. | Shen B. A new golden age of natural products drug discovery. Cell 2015;163:1297-300. |
| 4. | World Health Organization. Antimicrobial Resistance. Global Report on Surveillance. Geneva: World Health Organization; 2014. |
| 5. | Miroshnichenko A, Perfiliev V, Lysenko I, Zhernakova N. Interaction between some antibiotics and antioxidants. Res Result Pharmacol 2017;3:100-12. |
| 6. | Boyko NN, Zaytsev AI, Osolodchenko TP, Melnik, AL, Nevmerzhitskiy VV, Kazmirchuk VV. Antimicrobial activity of extracts from raw materials that contain phenolic compounds. Message 1. Pharmacom 2015;3:59-65. |
| 7. | Malyutina AY, Pravlotskaya AV, Novikov OO, Pisarev DI Study of the component composition of polyphenoles of the Kuril tea plant ( Pentaphylloides fruticosa L.). Pharm Pharm 2018;6:135-50. |
| 8. | Nefedova L, Boyko N, Zaytsev A, Osolodchenko T. Screening of antimicrobial properties of some tinctures. Pharm Kazakh 2013;11:42-4. |
| 9. | Lee HB, Kim JC, Lee SM. Antibacterial activity of two phloroglucinols, flavaspidic acids AB and PB, from Dryopteris crassirhizoma. Arch Pharm Res 2009;32:655-9. |
| 10. | Cermak P, Olsovska J, Mikyska A, Dusek M, Kadleckova Z, Vanicek J, et al. Strong antimicrobial activity of xanthohumol and other derivatives from hops ( Humulus lupulus L.) on gut anaerobic bacteria. APMIS 2017;125:1033-8. |
| 11. | Bharate SB, Bhutani KK, Khan SI, Tekwani BL, Jacob MR, Khan IA, et al. Biomimetic synthesis, antimicrobial, antileishmanial and antimalarial activities of euglobals and their analogues. Bioorg Med Chem 2006;14:1750-60. |
| 12. | Pal Singh I, Bharate SB. Phloroglucinol compounds of natural origin. Nat Prod Rep 2006;23:558-91. |
| 13. | Han X, Li Z, Li CY, Jia WN, Wang HT, Wang CH. Phytochemical constituents and biological activities of plants from the genus Dryopteris. Chem Biodivers 2015;12:1131-62. |
| 14. | Soliman FM, Fathy MM, Salama MM, Al-Abd AM, Saber FR, El-Halawany AM. Cytotoxic activity of acyl phloroglucinols isolated from the leaves of Eucalyptus cinerea F. Muell. Ex benth. Cultivated in Egypt. Sci Rep 2014;4:5410. |
| 15. | Buckwold VE, Wilson RJ, Nalca A, Beer BB, Voss TG, Turpin JA, et al. Antiviral activity of hop constituents against a series of DNA and RNA viruses. Antiviral Res 2004;61:57-62. |
| 16. | Takasaki M, Konoshima T, Kozuka M, Tokuda H. Anti-tumor-promoting activities of euglobals from Eucalyptus plants. Biol Pharm Bull 1995;18:435-8. |
| 17. | Lu C, Zhang HY, Ji J, Wang GX. In vivo anthelmintic activity of Dryopteris crassirhizoma, Kochia scoparia, and Polygala tenuifolia against Dactylogyrus intermedius (Monogenea) in goldfish ( Carassius auratus). Parasitol Res 2012;110:1085-90. |
| 18. | World Health Organization. Monographs on Selected Medicinal Plants. Vol. 2. Geneva: World Health Organization; 2002. |
| 19. | Duke JA, Bogenschutz-Godwin MJ, Duke PA. Handbook of Medicinal Herbs. 2 nd ed. Boca Raton: CRC Press LLC; 2002. |
| 20. | Coppen JJ. Eucalyptus: The Genus Eucalyptus. London and New York: CRC Press; 2002. |
| 21. | Popova NV, Litvinenko VI, Kucanjan AS. Medicinal Plants of the World: Encyclopedic Handbook. Kharkiv: Disa Plus; 2016. |
| 22. | Takasaki M, Konoshima T, Shingu T, Tokuda H, Nishino H, Iwashima A, et al. Structures of euglobal-G1, -G2, and -G3 from Eucalyptus grandis, three new inhibitors of epstein-barr virus activation. Chem Pharm Bull (Tokyo) 1990;38:1444-6. |
| 23. | Zhilyakova ET, Novikov OO, Pisarev DI, Malyutina AY, Boyko NN. Studying the polyphenolic structure of Laurus nobilis L. leaves. Indo Am J Pharm Sci 2017;4:3066-74. |
| 24. | European Pharmacopoeia. 8 th ed. Strasbourg: European Directorate for the Quality of Medicines and HealthCare; 2014. |
| 25. | Boyko NN, Makarevich NA, Pisarev DI, Zhilyakova ET. Simplified mathematical modeling of the distribution process of licuroside and glycyram between the extractant and Glycyrrhizae radices. Res Result Med Pharm 2018;4:75-80. |
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]
[Table 1], [Table 2], [Table 3]
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