[PL-04]
TRENDS AND ACHIEVEMENTS OF THE MEDICINAL AND AROMATIC PLANT RESEARCH IN BULGARIA

Stefan NICOLOV1, Elena GENOVA2 and Tatyana STOEVA2
1Department of Pharmacognosy and Botany, Faculty of Pharmacy, Dunav str. 2, 1000 Sofia, Bulgaria
2Institute of Botany, Bulgarian Academy of Sciences, Acad. G. Bonchev str. Bl. 23, 1113 Sofia, Bulgaria

Bulgaria is relatively small in size (111.000 km2) but it is rich in biological diversity - 7835 plant species: Algae - 3000, Lichenes - 710, Bryophyta - 660, Lycopodiophyta - 7, Equisetophyta - 12, Polypodiophyta - 30, Pinophyta - 16, Magnoliophyta - 3400 (55). In the practice of the folk medicine about 750 species, mainly vascular plants, are used. However, for the needs of pharmaceutical industry, phytoterapy and for export 200 species are applied more frequently. In the recent years the attention of the Bulgarian scientists is directed to four main issues:

  1. Study and protection of the genefund of medicinal and aromatic plants;

  2. Quantitative and qualitative analyses of the active substances in some rare and valuable species;

  3. Introduction and cultivation of the economically valuable plants;

  4. Standardisation and control of herbal drug raw material.

In 1995 "Chorological atlas of the medicinal plants in Bulgaria" was published, containing data about the chorology, ecological and biological characteristics, reproductive capacity, resources and measures for protection of 196 wild growing species. The biodiversity of the rare medicinal and aromatic plants and species with restricted regime of use was studied in the mountains of Znepole floral region (12), Rila National Park (76), and Central Balkan National Park (10). On the other hand, extensive biological and ecological investigations were carried out with some protected by law and rare species, such as Arctostaphylos uva-ursi (L.) Spreng. (58), Ruta graveolens L. (59) Sideritis syriaca L. (9), Alkanna tinctoria (L.) Tausch (15) etc. The problem of the studying and protection of some medicinal plants in the forest lands was also discussed (66). The biotechnological methods were applied for the reproduction of Leucojum aestivum L. (64,63), Atropa belladonna L. (5), Glycyrrhiza glabra L., Origanum heracleoticum etc. (72) in order their genefund to be protected. The genetic material from the protected and rare species is conserved in two ex situ collections: the Experimental Field of the Institute of Botany, BAS with 120 species (6), the Experimental Field of the Institute of Plant Genetic Resources, Sadovo with 38 species (72). Some of those investigations were financed by the National Research Foundation (NRF), Bulgarian Ministry of Environment and Waters (MEW), Global Environmental Fund (GEF) etc.

A study of the content and dynamics of accumulation of some active compounds in medicinal plants has been carried out in the period of 1990-2000:

Some methods for quantitative determination and separation of the active compounds have been applied:

  • a titrimetric method for total alkaloid determination in overground parts of Tribulus terrestris L. (Fabaceae) (50);

  • a densitometric method for quantitative determination of the xanthone isomers in Cratoxylum pruniflorum Kurz. and Hypericum aucheri Jaub. et Spach. (Guttiferae) (39) and

  • a method for separation of flavonoid and xanthone aglycones (28).

  • By using modern technics as HPLC, GC/MS, IR, UV, EI, 1H, 13C NMR etc., the qualitative analyses have been worked out for the following species.

    Essential oil in some aromatic plants:

    Active compounds in some medicinal plants:

    Phenolic acids, xanthones, catechins, flavonoids and related phenolic components:

    triterpenoid saponins: alkaloids:

    Some of the active compounds were successfully used as chemotaxonomic markers for the clarification of the taxonomy of some critical groups or families:

    The application of flavonoids, isoenzymes and pigments as markers in the phytomonitoring investigations has been also discussed (51,52,53,57). There are some data about the biological activity of some active compounds as the following:

    According to the recommendations of the Ministry of Environment and Waters (Regulation #973/1991) successful experiments with introduction and cultivation of some wild growing plants as Alchemilla species, Carlina acanthifolia All., Hippophae rhamnoides L., Sideritis scardica L. were carried out in this period.

    On the basis of the genetic material from wild growing and introduced species from abroad plant varieties were selected. The List of Varieties and Hybrids Permitted for Distribution in the Republic of Bulgaria (1995) includes 66 medicinal and aromatic plant varieties, achieved mainly of the Institute for Roses, Essential Oil and Medicinal Plants, Kazanluk; the Institute of Botany - BAS, Sofia; the High Institute of Agriculture, Plovdiv; the Institute of Plant Genetic Resources, Sadovo and the Institute of Plant Genetics - BAS, Sofia.

    Efforts of the Bulgarian scientists working in the field of a complex analysis of the standardization documents, which determine the quality of the herbal drugs in Bulgaria, have been made. Results and recommendations for development of an effective system for standardization and control of the herbal drug raw material, according to the recent scientific requirements are presented (47). By the pharmacognostical study of rhizoma cum radicibus and folium Urticae contemporary indexes for a new standard has been developed which should provide higher requirements for that herb as treating means and its realization on the international market (44,17).


    REFERENCES
    1. Angelova S., Varbanova K., Peeva J., Guteva Y. and Dimitrova D. (1994): Cultivation of medicinal plants from the wild flora in Bulgaria - possibilities and prospects. J. Herbs, Spices and Medicinal Plants 2, (2), 3-8.

    2. Benbassat N. and Nikolov St. (1995): Flavonoids from Astragalus onobrychis L., Planta medica, 61, 1, 100.

    3. Bondev I. (Ed.)(1995): Chorological Atlas of the Medicinal Plants in Bulgaria, Sofia, 272.

    4. Christov V., Simeonov M., Velcheva N., Karadjova O., Atanassov N., Ivanova I. and Evstatieva L. (1997): Pyrrolizidine alkaloids from Bulgarian species - genus Senecio and their insecticidal properties. Biotechnol. & Biotechnol. Eq. 11, (1), 53-59.

    5. Dimitrova D., Varbanova K. and Evstatieva L. (1996): Possibilities of in vitro propagation of Atropa belladona L. Proceedings Scientific Papers Second Balkan Scientific Conference on Study, Conservation and Utilisation of Forest Resources, 3-5. June 1996, Sofia. Sofia, 1, 385-388.

    6. Evstatieva L. (2000): Investigation of the threatened medicinal plants in Bulgaria, In: Dimitrova S. (Ed.), Medicinal plants - Solution 2000, 34-40.

    7. Evstatieva L. and Stoyanova K. (1994): Variability in the content of antraquinones in the rhizomes of Rumex alpinus L. Ann.de l'Univ. de Sofia "St.Kl.Ochridski" 2 - Botany, 85, 265-270.

    8. Evstatieva L., Todorova M.N., Ognyanov I.V. and Kuleva L.V. (1996): Chemical composition of the essential oil in Acorus calamus L. Fitologya 48, 19-25.

    9. Evstatieva L. and Vassilev P. (1995): Distribution, ecological peculiarities and phytocoenology of Sideritis syriaca L. in Bulgaria, In: Tsankov, G. (Ed.), Proceedings of Jubileum Symposium (2-3 June 1994) 100 Years from Birthday of the Acad. B. Stefanov (1894-1979), Sofia, 2, 181-186.

    10. Evstatieva L. and Vitkova A. (2000): Biodiversity of medicinal plants in the Central Balkan National Park, In: Sakalian M. (Ed.), Biological diversity of the Central Balkan National Park, Sofia, 58-105.

    11. Genova E. and Ivancheva S. (1995): Quantitative content of the tannins in Geranium macrorrhizum L. growing in Bulgaria, Phytologia Balcanica 1, 93-99.

    12. Genova E., Evstatieva L., Vitkova A., Gussev C., Stoeva T. and Peshevski N. (1996): Mapping of the medicinal plants in some mountains of the Znepole floral region, In: Tsankov G. (Ed.), Proceeding Scientific Papers Second Balkan Scientific Conference on Study, Conservation and Utilization of Forest Resources (3-5 June, 1996, Sofia), Sofia, 1, 367-373.

    13. Genova E., Hristova J. and Beeva J. (1998): Study on the content of some biologically active compounds in Liquorice (Glycyrrhiza glabra L.) Pharmacia 3-4, 10-13.

    14. Genova E., Kitanov G. and Stefanova I. (1995): Study on the alkannin (shikonin) content in the roots of dyer's alkanet (Alkanna tinctoria (L.) Tausch.) Pharmacia 3, 15-17.

    15. Genova E., Rusakova V. and Stefanova I. (1997): On the distribution, ecobiological and phytocoenotic specificities of Alkanna tinctoria (L.) Tausch. in Bulgaria. Phytologia Balcanica 3/2-3, 167-173.

    16. Genova E., Terzieva L., Beeva J., Nikolov S. and Thuan N. (1995): Biological and phytochemical studies of Platycodon grandiflorus (Jacq.) A. DC. introduced in Bulgaria. In: Tsankov G. (Ed.), Proceeding of Jubileum Symposium (2-3 June '94) 100 Years from Birthday of the Acad. B. Stefanov (1894-1979), Sofia, 2, 209-212.

    17. Gussev Ch. and Nikolov St. (1995): Standartization of Folium Urticae, Bulletin of the Maced. Pharmac. Assoc. 41, 1-2, 396-397.

    18. Handjieva N., Popov S. and Evstatieva L. (1996): Constituents of Essential oils from Nepeta cataria L., N. grandiflora M.B. and N. nuda. J. Essent.Oil Res. 8, 1-5.

    19. Ivancheva S., Genova E. and Taleva R. (1994): Dynamics of polyphenols in the Geranium sanguineum L. Ann. de l'Univ. de Sofia, Faculte de Biologie, 85, (2), 271-275.

    20. Ivancheva S. and Nedelcheva A. (1995): Flavonoids in genus Achillea- sect. Filipendulinae (Asteraceae). Polyphenols 94. In: Brouillard R., Jay M., Scalbert A. (Eds), 17th International Conference on Polyphenols, Palma de Mallorca (Spain), May 23-27 1994. Les Colloques, N 69, INRA, Paris, 303-304.

    21. Ivancheva S. and Nedelcheva A. (1997): Exudate flavonoid aglycones of Achillea sp. sect. Millefolium and sect. Filipendulinae - a comparative study. Zbornik radova sa V. Simpozijuma o flori jugoistočne Srbije, Zaječar, 183-188.

    22. Ivancheva S. and Petrova A. (2000): Chemotaxonomic study of eleven Geranium species Biochemical Systematics and Ecology. 28, 255-260.

    23. Ivancheva S., Stojanov D., Stancheva B. and Cherneva Z. (1998): Polyphenol compounds in genus Vaccinium, (Ericaceae). Pharmacia XLV, 3-4, 7-9.

    24. Ivancheva S. and Vitkova A. (1997): Flavonoids in Solidago virgaurea L. Pharmacia XLIV, 1, 17-19.

    25. Kitanov G.M. (1992): Phenolic acids and flavonoids from Stellaria media (L.) Vill. (Caryophyllaceae). Pharmazie. 47, 470-471.

    26. Kitanov G.M. (1995): Dynamics and contents of tannins and flavonoids in Hypericum maculatum Grantz. Farmacia (Sofia). 43 (5-6), 13-17.

    27. Kitanov G.M. (1995): Phytochemical investigation of Potentilla reptans. Bulletin of the Macedonian Pharmaceutical Association. 41, 392-393.

    28. Kitanov G.M. (2000): A method for separation of flavonoid and xanthone aglycones with mono hydroxyphenyl and o-dihydroxyphenyl moiety. Acta Pharm. 50, 69-73.

    29. Kitanov G.M. (2000): The dynamics and contents of hypericin in Hypericum perforatum L. and Hypericum maculatum Grantz. Growing in Bulgaria. Acta Pharm. 50, 65-68.

    30. Kitanov G.M., Dam The Van and Assenov I. (1991): Chemical composition of Gentiana asclepiadea roots, Khim. Prir. Soedin. 27, 425-427.

    31. Kitanov G.M., Dam The Van and Assenov I. (1992): Flavonols from Melaleuca leucadendron leaves. Fitoterapia. 63, 379-380.

    32. Kitanov G.M. and Nedialkov T.T. (1998): Mangiferin and isomangiferin in some Hypericum species. Biochem. Syst. Ecol. 26, 647-653.

    33. Kitanov G.M. and Nikolov N.Tz. (1991): Phenolic constituents of Hypericum maculatum, Fitoterapia. 62, 92.

    34. Kitanov G.M. and Pashankov P. (1994): Quantitative investigation on the dynamics of plumbagin in Plumbago europaea L. roots and herb by HPLC. Pharmazie. 49, 462.

    35. Kitanov G.M. and Spassov S.I. (1992): Naphtodipyranodione from Gentiana asclepiadea. Phytochemistry. 31, 1067-1068.

    36. Kulevanova S., Stoeva T. and Ristić M. The essential oil composition of Thymus tosevii and T. macedonicus from Bulgaria. Bollettino Chimico Farmac. (in press).

    37. Marinova S., Stoichkov J., Benbassat N., Nikolov St. and Surneva M. (1994): Immuno-modulating effect of triterpenoid saponins from Astragalus onobrychis L. Farmacia. 42, 4, 38-41.

    38. Nedelcheva A. and Ivancheva S. (1997): Quantitative study of flavonoid content in species from genus Achillea L., sect. Filipendulinae (DC.) Afan. (Asreraceae). Annuaire de l'Universite de Sofia "St. Kliment Ohridski", 6 eme session scientifique, Sofia'96. 88, (4), 101-105.

    39. Nedialkov T.T., Kitanov G.M. and Tencheva J. (1998): Densitometric determination of the xanthone isomers mangiferin and isomangiferin in plant materials. Acta Pharm. 48, 211-214.

    40. Nguyan Thuan and Nikolov St. (1995): Phytochemical studies on Platycodon grandiflorus (Jacq) A.CD., introduced in Bulgaria, J.Pharm.Univ. of Marmara (Turkey). 11, 1-2, 351-355.

    41. Nikolov St. and Benbassat N. (1995): A triterpenoid saponin from Astragalus onobrychis L., Bulletin of the Maced. Pharmac. Assoc. 41, 1-2, 398-399.

    42. Nikolov St. and Benbassat N. (1995): Flavonoids from Astragalus vesicarius L., Pharmacia. 43, 2, 4-5.
    43. Nikolov St. and Benbassat N. (1997): Triterpenoid saponins and sapogenins from genus Astragalus L., Farmacia, 44, 3-4, 34-48.

    44. Nikolov St. and Gussev Ch. (1992): Determination of quantitative characteristics of rhizome and root of the drug Urtica (Rhizoma cum radicibus Urticae), Biotehnologia & biotehnika. 6, 1, 52-55.

    45. Nikolov St. and Gussev Ch. (1997): A chemotaxonomic comparation of genera Asparagus and Ruscus (Liliaceae), Bocconea, 5, 2, 765-770.

    46. Nikolov St. and Gussev Ch. (1997): The dynamics of acumulation of ruscogenin in the roots and rhizomes of Ruscus aculeatus (Liliaceae), Acta Pharm. 47, 3, 203-206.

    47. Nikolov St., Gussev Ch., Kitanov G. and Assenov I. (1994): Standardization of herbal drugs in Bulgaria - existing situation, problems and present-days requirements, Farmacia. 42, 1-2, 49-54.

    48. Nikolov St., Thuan N. and Zheljazkov V. (1996): Flavonoids from Achyranthes bidentata BC., Acta Horticulture. 426, 75-78.

    49. Nikolova M., Nikolov S. and Koeva Y. (1998): Flavanoids in the overground part of Pulsatilla slavianke (Rumm.) D. Yord. et Koz. (Ranunculaceae), Phytologia Balcanica. 4, 1-2, 55-59.

    50. Obreshkova D., Mitkov S. and Pangarova T. (1999): Alkaloid of Tribulus terrestris L. I. Quantitative determination of total alkaloid content, Pharmacia. 46, 1, 19.

    51. Peev D., Ivancheva S., Angelov G., Kurteva M. and Derebanova P. (1995): Phytomonitoring in Rila mountain- 1994. II. Flavonoids, isoenzymes and pigments of the control populations. Observatiore de montagne de Moussala, 3, 162-171.

    52. Peev D., Ivancheva S., Angelov G., Kurteva M. and Delcheva M. (1996): Phytomonitoring in Rila Mountain- 1995. II. Flavonoids, isoenzymes and pigments of the control populations. Observatoire de montagne de Moussala OM2, 4, 153-162.

    53. Peev D., Ivancheva S., Angelov G., Kurteva M. and Delcheva M. (1997): Phytomonitoring in Rila Mountain- 1996. II. Flavonoids, isoenzymes and pigments of the control populations. Observatoire de montagne de Moussala OM2, 6, S, 106-114.

    54. Peneva B., Krasteva I., Nikolov St. and Nedialkov P. (2000): In vitro release of ruscogenins from suppositories with dry extract of Ruscus aculeatus L. In: 3rd World Meeting on Pharmaceutics Biopharmaceutics Pharmaceutical Technology, Berlin, 3-6 april. 817-818.

    55. Petrova A., Anchev M. and Palamarev E. (1999): Ekskurzionen opredelitel, Sofia, 837.

    56. Philipov S., Evstatieva L. and Michailova M. (1995): Comparative phytochemical study of the alkaloid content of Berberis vulgaris L. Phytologia Balcanica 2, 75-80.

    57. Renke K. and Ivancheva S. (1996): Effect of environment pollution on the chemical composition of medicinal plants. Chemia i inzyneria ekologiczna, 3, (1), 119-123.

    58. Rusakova V., Genova E. and Cherneva Z. (1991): Phytocoenological and ecological peculiarities of the bear berry (Arctostaphylos uva-ursi (L.) Spreng.) in some regions of Bulgaria, Fitologya 39, 33-53.

    59. Rusakova V. and Vitkova A. (1995): Phytocoenological characteristics of Ruta graveolens L. in Bulgaria. Phytologia Balcanica 1, 39-57.

    60. Serkedjieva J. and Ivancheva S. (1996): Antiviral activity of species in Geraniaceae. Polyphenols Communications 96, Bordeaux (France), July 15-18. 2, 449-450.

    61. Serkedjieva J. and Ivancheva S. (1998): Antiviral effects of extracts from Geranium sanguineum L. Polyphenol Communications 98, XIX International Conference on Polyphenols, Lille (France) 1-4 September 1998, 125-126.

    62. Serkedjieva J. and Ivancheva S. (1999): Antiherpes virus activity of extracts from the medicinal plant Geranium sanguineum L. Journal of Ethnopharmacology 64, 59-68.

    63. Stanilova M. (1994): In vitro regeneration of Leucojum aestivum L. from leaf explants, Bulg. J. of Plant Physiol. 20/1-4, 88-94.

    64. Steffanov Z. (1990): Ecologo-biologichni i fitohimichni prouchvanija varhu estestvenite populacii i introducirani proizhodi na blatnoto kokiche (Leucojum aestivum L.) v Bulgaria, Sofia, Doct. Thesis.

    65. Stoeva T., Ivancheva St. and Behar M. (1997): Flavonoids, phenolic and triterpenic acids in Mentha spicata group from Bulgaria. Phytologia Balcanica 3/1, 89-93.

    66. Stojanova M. (1995): Conservation and utilization of the medicinal plants in forest lands. In: Tsankov G. (Ed.), Proceedings Scientific Papers, Jub. Simpos. 125 Years BAS and 65 Years Forest Research Institute, 22-23. September 1994, Sofia, 122-125.

    67. Taskova R. (1999): Sravnitelno chemotaxonomichno prouchvane na vidove ot rodovete Veronica s. l. (Scrophulariaceae) i Plantago L. (Plantaginaceae), Sofia, Doct. Thesis.

    68. Taskova R., Mitova M., Evstatieva L., Anchev M., Peev D., Handjieva N., Bankova V. and Popov S. (1997): Iridoids, flavonoids and terpenoids as taxonomic markers in Lamiaceae, Scrophulariaceae and Rubiaceae. Bocconea 5 (2), 631-636.

    69. Todorova M., Kuleva L., Christov R. and Vitkova A. (1997): Chemotypes of Artemisia lerchiana Weber. From Bulgaria. J. Essent. Oil Res. 9, 299-301.

    70. Tsankova E., Konakchiev A. and Genova E. (1993): Chemical Composition of the Two Hyssopus officinalis Taxa. J. Essential Oil Res. 5, 609-611.

    71. Tsankova E., Konakchiev A. and Genova E.: (1994): Constituents of essential oils from three Salvia species, J. Ess. Oil Res. 6, 375-378.

    72. Varbanova K. and Dimitrova D. (2000): Gathering, reproduction and preservation of species of wild-growing medicinal plants, In: Dimitrova S. (Ed.), Medicinal plants - Solution 2000, 41-55.

    73. Velev G., Ivancheva S. and Bourzeix M. (1995): Antiradiation effect of polyphenols from Geranium sanguineum L. Polyphenols 94. In: Brouillard R., Jay M., Scalbert A. (Eds.), 17th International Conference on Polyphenols, Palma de Mallorca (Spain), May 23-27 1994. Les Colloques, N 69, INRA, Paris, 401-402.

    74. Vitkova A. (1996): A comparative assessment and dynamics of accumulation of flavonoids and tannins in species of Alchemilla L.(Rosaceae). Fitologya 48, 11.

    75. Vitkova A. (1999): Ekologichno i fitohimichno proucvane na Ruta graveolens L. v Bulgaria, Sofia, Doct. Thesis.

    76. Vitkova A. and Evstatieva L. (2000): Biodiversity of medicinal plants in the Rila National Park, In: Sakalian M. (Ed.), Biological diversity of the Rila National Park, Sofia, 79-116.

    [PL-04]