Postępy
nasze czasopisma medyczne
New Medicine
Postępy Nauk Medycznych
Medycyna Rodzinna
Nowa Medycyna
Nowa Pediatria
Nowa Stomatologia
oferta Wydawnictwa Borgis
CzytelniaMedyczna.pl
Księgarnia Medyczna Borgis.pl
Księgarnia ogólna DoPoduszki.pl
Newsletter Biuletyn Telegram
Kosmetyki ziołowe
Postępy
Wydawca:
Wydawnictwo Medyczne Borgis

Organ
Sekcji Fitoterapii PTL
Sekcja Fitoterapii Polskiego Towarzystwa Lekarskiego

Rodzaj Panax – systematyka, skład chemiczny, działanie i zastosowanie oraz analiza fitochemiczna nadziemnych i podziemnych organów żeń-szenia amerykańskiego – Panax quinquefolium L. Metody ekstrakcji i analizy ginsenozydów. Cz. II.

© Borgis - Postepy Fitoterapii 3, s. 139-164
*Tadeusz Wolski, Agnieszka Ludwiczuk, Tomasz Baj, Kazimierz Głowniak
Rodzaj Panax – systematyka, skład chemiczny, działanie i zastosowanie oraz analiza fitochemiczna nadziemnych i podziemnych organów żeń-szenia amerykańskiego – Panax quinquefolium L. Metody ekstrakcji i analizy ginsenozydów. Cz. II.
Genus panax – taxonomy, chemical composition, pharmacological effects, medicinal application and phytochemical analysis of aerial and underground parts of american ginseng (panax quinquefolium l.). method of extraction and determination of ginsenosides. part ii
Katedra i Zakład Farmakognozji z Pracownią Roślin Leczniczych, Uniwersytet Medyczny, Lublin
Kierownik Katedry i Zakładu: prof. dr hab. Kazimierz Głowniak
Summary
In this article literature review regarding extraction and identification methods of ginsenosides occurring in Panax genus was presented. Own data which concern analysis of ginsenosides fraction from Panax quinquefolium L. was also described. The occurrence of these compounds in plant material collected in different periods of ginseng vegetation as well in pharmaceutical preparations received from Panax ginseng was compared. This article contain data concerning preliminary investigations of ginseng e.g. the loss of drying and also quantitative analysis of ginsenosides by spectrophotometric method, optimization of extraction method and chromatographic separation for TLC, HPTLC, OPLC and HPLC methods. The quantitative analysis of the main compounds using TLC and HPLC methods in optimized chromatographic conditions was also performed.
Preliminary investigations concerning the loss of drying showed that underground parts of ginseng are characterized by higher content of moisture in comparison to aerial parts. The content of ginsenosides in roots as well in aerial parts of P. quinquefolium increased along with the age of the plants. The highest yield of ginsenosides was obtained in 4-year-old leaves and roots of P. quinquefolium. Ginseng leaves in comparison to ginseng roots are characterized by highest concentration of ginsenosides. Percentage of ginsenosides in roots is ranged from 6.5 to 12.5% while in ginseng leaves from 24.8 to 37.5%. Stems and fruits of Panax quinquefolium are characterized by much lower content of saponins.
Studies on isolation of ginsenosides were concerned selection of extraction solvent and extraction method. The highest extraction efficiency of ginsenosides in roots and leaves of American ginseng was observed during mechanical shaking with 50% aqueous methanol. Separation of ginsenosides conducted in 26°C and by use of chloroform – methanol – ethyl acetate – water, 15+22+40+9 (v/v) as the mobile phase was characterized by good selectivity and resolution of saponins. The best results for ginsenosides separation, using OPLC method, gave mobile phase: chloroform – methanol – ethyl acetate – water – hexane, 20+22+60+8+4 (v/v). Results from TLC, HPTLC and OPLC analyses of ginsenosides indicated that use of the different development methods resulted in significant differences between the resolutions of the compounds. Forced-flow OPLC is more sensitive method for analysis of ginsenosides.
Quantitative analysis of the main ginsenosides occurring in leaves and roots of American ginseng was executed using TLC and HPLC methods in optimized chromatographic conditions. The linear calibration curves for 6 ginsenosides, concentration ranges, limits of detection and limits of quantification for both TLC and HPLC was determined. Recovery for all investigated ginsenosides was between 88.8 and 101.2%. TLC and HPLC analyses showed that the main compound of American ginseng roots is ginsenoside Rb1. The amount of this compound was ranged from 1.36 to 3% according to extraction method and extraction solvent used. Ginsenosides Rd, Rg2 and Rb2 are the main compounds occurring in ginseng leaves. They content in plant material after mechanical shaking was 3.8, 2.4 and 1.4% respectively. The amount of another ginsenosides in ginseng leaves not exceeded 1%. Ginseng leaves, in comparison to ginseng roots are characterized by higher concentration of ginsenosides. Therefore, based on the concentration of major saponins, leaves can by alternative to root source of ginsenosides used in herbal preparations. The lack of ginsenoside Rf in aerial and undergrounds parts of Panax quinquefolium confirmed its status as a phytochemical marker differentiating American and Asian ginseng.
Key words: panax quinquefolium, american ginseng, ginsenosides, qualitative analysis, quantitative analysis, extraction methods, uae, ase, tlc, oplc
Piśmiennictwo
1. Wolski T, Ludwiczuk A, Baj T i wsp. Rodzaj Panax - systematyka, skład chemiczny, działanie i zastosowanie oraz analiza fitochemiczna nadziemnych i podziemnych organów żen-szenia amerykańskiego - Panax quinquefolium L.). Cz. I. Postępy Fitoterapii 2008; 2:96-114. 2. Ando T, Tanaka O, Shibata S. Chemical studies on the oriental plant drugs. XXV. Comparative studies on the saponins and sapogenins of ginseng and related crude drugs. Svoyakugaku Zasshi 1971; 25:28-32. 3. Chuang WC, Sheu SJ. Determination of ginsenosides in ginseng crude extracts by high-performance liquid chromatography. J Chromatogr A 1994, 685:243-51. 4. Wu J, Lin L, Chau F. Ultrasound-assisted extraction of ginseng saponins from ginseng roots and cultured ginseng cells. Ultrason Sonochem 2001; 8:347-52. 5. Choi JH, Kim DH, Sung HS i wsp. Kinetic studies on the thermal degradation of ginsenosides in ginseng extract. Korean J Food Sci Tech1982; 14(3):197-202. 6. Sung HS, Yang JW. Effect of the heating treatment on the stability of saponin in white ginseng. J. Korean Society of Food and Nutrition 1986; 15:22-6. 7. Mason TJ, Paniwnyk L, Lorimer JP. The uses of ultrasound in food technology. Ultrason Sonochem 1996; 3:253-60. 8. Giergielewicz-Możajska H, Dąbrowski Ł, Namieśnik J. Przegląd technik ekstrakcyjnych wykorzystywanych na etapie wyodrębniania analitów z próbek stałych. Ekologia i Technika 2001; 1(49):3-11. 9. Choi MPK, Chan KKC, Leung HW i wsp. Pressurized liquid extraction of active irgendients (ginsenosides) from medicinal plants using non-ionic surfactant solutions. J Chromatogr A, 2003; 983:153-62. 10. Lopez-Avila V, Young R, Teplitsky N. (1996), Microvawe-assisted extraction as an alternative to Soxhlet, sonication and supercritical fliud extraction. Journal of the Association of Official Analytical Chemists International 1996; 79(1):142-56. 11. Kwon JH, Belanger JMR, Pare JRJ i wsp. Application of the microwave-assisted process (MAPTM) to the fast extraction of ginseng saponins. Food Research International 2003; 36:491-8. 12. Wolski T, Ludwiczuk A. (2001), Ekstrakcja produktów naturalnych gazami w stanie nadkrytycznym. Przem Chem 2001; 80 (7):286-9. 13. Wang J, Sha Y, Li W i wsp. Quinquenoside L9 from leaves and stems of Panax quinquefolium L. J Asian Nat Prod Res 2001; 3(4):293-7. 14. Sticher O, Soldati F. HPLC Trennung und quantitative Bestimmung der Ginsenoside von Panax ginseng, Panax quinquefolium und von Ginseng-Spezialitäten. Planta Med 1979; 36:30-42. 15. Glockl I, Veit M, Blaschke G. Determination of ginsenosides from Panax ginseng using micellar electrokinetic chromatography. Planta Med 2002; 68(2):158-61. 16. Hiai S, Oura H, Hamanaka H i wsp. A color reaction of panaxadiol with vanillin and sulphuric acid. Planta Med 1975; 28:131-8.17. Hiai S, Oura H, Odaka Y i wsp. A colorimetric estimation of ginseng saponins. Planta Med 1975a; 28:363-9. 18. Farmakopea Niemiecka DAB 10, Ginseng radix, 1991. 19. Hostettmann K, Terreaux C, Marston A i wsp. The role of Planar Chromatography in the rapid screening and isolation of bioactive compounds from medicinal plants. J Planar Chromatogr 1997; 10:251-7. 20. Corthout J, Naessens T, Apers S i wsp. Quantitative determination of ginsenosides from Panax ginseng roots and ginseng preparations by thin layer chromatography-densitometry. J Pharm Biomed Anal 1999; 21:187-92. 21. Vanhaelen-Fastre RJ, Faes ML, Vanhaelen MH. High-performance thin-layer chromatographic determination of six major ginsenosides in Panax ginseng. J Chromatogr A 2000; 868:269-76. 22. Nyiredy Sz. Rotation Planar Chromatography. In: Sz. Nyiredy (Editor) Planar Chromatography. A retrospective view for the third millennium. Springer, Budapest, 2001; 178-99. 23. Ludwiczuk A, Nyiredy Sz, Wolski T. Separation of the ginsenosides fraction obtained from the roots of Panax quinquefolium L. cultivated in Poland. J Planar Chromatogr 2005; 2:104-7. 24. Ludwiczuk A, Kołodziej B, Wolski T. Zawartość i skład ginsenozydów w różnych organach żeń-szenia pięciolistnego ( Panax quinquefolium L.) Acta Agrobot 2006; 59(1):507-14. 25. Ligor T, Ludwiczuk A, Wolski T, Buszewski B. Isolation and determination of ginsenosides in American ginseng leaves and root extracts by LC-MS. Anal Bioanal Chem 2005; 383(7-8):1098-105. 26. Wang X, Sakuma T, Asafu-Adjaye E i wsp. Determination of ginsenosides in plant extracts from Panax ginseng and Panax quinquefolium by LC/MS/MS. Anal Chem 1999; 71(8):1579-84. 27. Shangguan D, Han H, Zhao R i wsp. New method for high-performance liquid chromatographic separation and fluorescence detection of ginsenosides. J Chromatogr A 2001; 910:367-72. 28. Cui JF. Identification and quantification of ginsenosides in various commercial ginseng preparations. European J Pharm Sci 1995; 3:77-85. 29. Cui JF, Bjorkhem I, Eneroth P. Gas chromatographic - mass spectrometric determination of 20(S)-protopanaxadiol and 20(S)-protopanaxatriol for study on human urinary excretion of ginsenosides after ingestion of ginseng preparations. J Chromatogr B 1997; 689:349-55. 30. Kubo M, Tani T, Katsuki T i wsp. (1980), Histochemistry. I. Ginsenosides in ginseng ( Panax ginseng C.A. Meyer). J Nat Prod 1980; 43(2):278-84. 31. Segiet-Kujawa E, Lutomski J. Comparison of analytical methods of determining saponins in some Araliaceae species. Herba Pol 1986; 32(1):39-46. 32. Sticher O. Getting to the root of ginseng. CHEMTECH 1998; 28(4):26-32. 33. Dou DQ, Hou WB, Chen YJ. Studies on the characteristic constituents of Chinese ginseng and American ginseng. Planta Med 1998; 64:585-6. 34. Ludwiczuk A, Wolski T, Berbeć S. Chromatographic analysis of ginsenosides occurring in roots of American ginseng ( Panax quinquefolium L.) and Asian ginseng ( Panax ginseng C.A. Mayer) preparations. J Planar Chromatogr 2002; 15:147-50. 35. Dallenbach-Toelke K, Nyiredy Sz, Meszaros SY i wsp., Sticher O. TLC, HPTLC and OPLC separation of ginsenosides. J High Res Chromatogr & Chromatogr Comm 1987; 10:362-4. 36. Soldati F, Sticher O. HPLC separation and quantitative determination of ginsenosides from Panax ginseng, Panax quinquefolium and from Ginseng Drug Preparations. Planta Med 1980; 38:346-57. 37. Court WA, Hendel JG, Elmi J. Reversed-phase high-performance liquid chromatographic determination of ginsenosides of Panax quinquefolium. J Chromatogr A 1996; 775:11-7. 38. Kwon SW, Han SB, Park IH i wsp. Liquid chromatographic determination of less polar ginsenosides in processed ginseng. J Chromatogr A 2001; 921:335-9. 39. Park MK, Kim BK, Park JH i wsp. High-performance liquid-chromatographic determination of ginsenosides using photoreduction fluorescence detection. J Liq Chromatogr, 1995; 18(10):2077-88. 40. Samukawa K, Yamashita H, Matsuda H i wsp. Simultaneous analysis of ginsenosides of various Ginseng radix by HPLC. Yakugaku-Zasshi 1995; 115(3):241-9. 41. Zhou Z, Zhang G. Analysis of ginseng. IV. HPLC determination of ginsenosides in Panax ginseng. Youxue-Xuebao 1988; 23(2):137-41. 42. Park MK, Park JH, Han SB i wsp. High-performance liquid-chromatographic analysis of ginseng saponins using evaporative light-scattering detection. J Chromatogr A 1996; 736(1-2): 77-81. 43. Ji HY, Lee HW, Kim HK i wsp. Simultaneous determination of ginsenoside Rb1 and Rg1 in human plasma by liquid chromatography - mass spectrometry. J Pharm Biomed Anal 2004; 35:207-12. 44. Lau AJ, Woo SO, Koh HL. Analysis of saponins in raw and steamed Panax notoginseng using high-performance chromatography with diode array detection. J Chromatogr A 2003; 1011:77-87. 45. Tani T, Kubo M, Katsuki T i wsp. Histochemistry. II. Ginsenosides in ginseng ( Panax ginseng, root). J Nat Prod 1981; 44(4):401-7. 46. Farmakopea Polska V, tom 1, Supl. PTFarm, Warszawa, 1995. 47. Farmakopea Polska VI, PTFarm, Warszawa, 2002. 48. Metoda Badawcza Nr MB:05F1100202, ZZ Herbapol w Klęce S.A.: Oznaczanie zawartości ginsenozydów w preparacie Extractum Ginseng spir. spiss. 49. Ludwiczuk A. Badania składu chemicznego w ontogenezie żeń-szenia amerykańskiego Panax quinquefolium L.. Praca doktorska. Uniwersytet Medyczny, Lublin, 2005. 50. Ludwiczuk A, Wolski T. Estimation of the content and composition of ginsenosides occurring in extracts from American ginseng and Asian ginseng. Annales UMCS, sec. EEE 2003; 7:53-8. 51. Ouyang MA. Glycosides from the leaves of Ilex latifolia. Chinese J Chem 2001; 19:885-92. 52. Strzelecka H, Kamińska J, Kowalski J i wsp. Chemiczne metody badań roślinnych surowców leczniczych. PZWL, Warszawa, 1987; pp. 312. 53. Nguyen TN. Study on Panax vietnamensis Ha et Grushv. - Araliaceae. Botany - tissue culture. Chemistry - biological properties. Herba Pol 1989; 35, Supl.II: pp.229. 54. Jerzmanowska Z. Substancje roślinne. Metody wyodrębniania. Tom II. PWN, Warszawa, 1970; pp. 200. 55. Gudej J, Tomczyk M, Urban E i wsp. Badania składu chemicznego liści Rubus saxatilis L. Herba Pol 1998; 44 (4):340-4. 56. Sieńko A. Dobór warunków ekstrakcji i rozdziału chromatograficznego ginsenozydów żeń-szenia amerykańskiego Panax quinquefolium L. Praca magisterska, Uniwersytet Medyczny w Lublinie, 2003; 84pp. 57. Ziewiec A. Badania nad składem chemicznym części nadziemnych i podziemnych żeń-szenia amerykańskiego Panax quinquefolium L. Praca magisterska, Uniwersytet Medyczny w Lublinie, 2002; 87pp . 58. Ludwiczuk A, Wolski T, Nyiredy Sz. Circular and linear OPLC of ginsenosides in Panax quinquefolium L. cultivated in Poland. Proceedings of the International Symposium on Planar Separations, Budapeszt, 21-23 czerwiec 2003, Węgry, 291-6. 59. Farmakopea Europejska, European Pharmacopoeia - Supl. 2001; 887-9. 60. Tyihak E, Mincsovics E. Forced-flow Planar Liquid Chromatographic Techniques. J Planar Chromatogr 1988; 1:6-19. 61. Tyihak E, Mincsovics E. Trends in Overpressured Layer Chromatography. J Planar Chromatogr 1991; 4:288-92. 62. Tyihak E, Mincsovics E. Overpressured-layer chromatography (optimum performance laminar chromatography). In: Sz. Nyiredy (Editor) Planar Chromatography. A retrospective view for the third millennium. Springer, Budapest, 2001; 137-76. 63. Nyiredy Sz. The bridge between TLC and HPLC: overpressured layer chromatography (OPLC). Trends in Analytical Chemistry 2001; 20 (2):91-101. 64. Nyiredy Sz. Planar chromatographic method development using the PRISMA optimization system and flow charts. J Chromatogr Sci 2002; 40:1-11. 65. Nyiredy Sz, Fater Zs, Botz L i wsp. The role of chamber saturation in optimization of planar chromatography. J Planar Chromatogr 1992; 5:308-15. 66. Dallenbach-Toelke K, Sticher O. Comparison of Circular and Linear Overpressured Layer Chromatography (OPLC). J Planar Chromatogr 1988; 1:73-5. 67. Szabady B. The different modes of development. In: Sz. Nyiredy (Editor). Planar Chromatography. A retrospective view for the third millennium. Springer, Budapest, 2001; 88-102. 68. Nyiredy Sz, Meszaros SY, Dallenbach-Toelke K i wsp. The "Disturbing Zone" in Overpressure Layer Chromatography (OPLC). J High Res Chromatogr and Chromatogr Comm 1987; 10:352-6. 69. Shibata S, Tanaka O, Shojii J i wsp. Chemistry and pharmacology of Panax. In H. Wagner, H. Hikkino and N.R. Farnsworth (Eds.) Economic and Medicinal Plant Research, Vol.1. Orlando, Fla: Academic Press, 1985; 218-84.

otrzymano/received: 2008-08-21
zaakceptowano/accepted: 2008-09-05

Adres/address:
*Tadeusz Wolski
Katedra i Zakład Farmakognozji z Pracownią Roślin Leczniczych Uniwersytetu Medycznego w Lublinie
ul. Chodźki 1, 20-093 Lublin
tel.: (0-81) 742-38-10, fax: (0-81) 742-38-09
e-mail: twolski@pharmacognosy.org
Copyright © Wydawnictwo Medyczne Borgis 2006-2013
Chcesz być na bieżąco? Polub nas na Facebooku: strona Wydawnictwa na Facebooku
do góry strony