Characterization of the flavonolignans in Silybummarianum L. Grown Naturally in Iraqi-Kurdistan Region


Mohammed NawzadSabir, Ali AbdulrahmanAskari

School of Pharmacy, University of Sulaimani 



Abstract: 

 (Abstracted from MSc. Thesis entitles "Pharmacognostic and Pharmacologic study of Silybum marianum grown naturally in Iraqi-Kurdistan Region") 
The antihepatotoxic principles found in the indigenous plant Silybum marianum L. (Milk thistle) was obtained using a multi-extraction/ separation and isolation procedures. Starting with identification of the plant and collection of the seeds. A pretreatment (defatting) step with light petroleum ether preceded the extraction with organic solvent (ethanol) in a soxhlet extractor. The alcoholic extract after drying was partitioned with n-hexane, Chloroform, Ethyl acetate, and n-butanol successively. Each fraction was separated by flash column chromatography. The sub-fractions after column chromatography were tested with thin layer chromatography (TLC) for the presence of the flavonolignans. Fractions showed spots with the same Rf-values were combined and furthermore isolated with high performance liquid chromatography (HPLC). Components were then purified and identified using UV, IR-spectroscopy. The results were compared with a standard sample of silybin (obtained from Sigma-Aldrich Co.). Results confirmed that the plant Silybum marianum L. which grows indigenous in Kurdistan Region/ Iraq, reserves the same active components (flavonolignans) present in the plant that grows elsewhere. 


References 
1-Bahl A., Bahl B.S.; A text book of Organic chemistry.18th ed 2008; S CHAND; 81-219-2614-9; pp: 7-20, 172-188. 347, 426. 
2-Prakash A. K., Lata S. K., Sudeep T., Kumar S.,; Invention of a novel process for the isolation of a hepatoprotective agent Silymarin from    the seeds of the plant Silybum marianum; Oct. 30, 2001; A patent number 6309678. 
3-Barreto J. F., Wallace S. N., Carrier D. J., Clausen E. C.; Extraction of nutraceuticals from milk thistle: II. Extraction with organic solvents; J of  Applied Biochem and Biotech; 2003; (105-108); 891-903. 
4-Benthin B., Danz H., Hamburger M.; Pressurized liquid extraction of medicinal plants; J of Chromatogr A.; 1999; (837); 211-219. 
5-Bhat S.V., Nagasampagi B.A., Sivakumar M.; Chemistry of natural products; 1st ed 2007; Springer, Berlin; 3-540-40669-7; (1-14), 585-  605. 
6-Klaassen C., Watkins III J. B.; Casarett & Doull’s Essentials of Toxicology, 2nd ed 2003; Mc Graw Hill, Lange; 978-0071622400; pp: 87, 199,  202, 366. 
7-Kroll David J., Shaw H. S., Oberlies N. H.; Milk thistle nomenclature: Why it matters in Cancer research and Pharmacokinetic studies; J of  Integr Cancer Ther. 2007; (6); 110.  
8-Stahl E.; Thin layer chromatography; 2nd ed 1988; Springer-Verlag Berlin; 08247-0895-4; 268-272. 
9-Ahmed I., Malik A., Ferheen S., Afza N., Muhammad A., Lodhi A., Choudhary M. I.; Chymotrypsin Inhibitory Triterpenoids from Silybum  marianum; Chem and Pharmacol Bull; 2006; (54) 1 :103-106. 
10-Loudon G. M.,; Organic chemistry; 3rd ed Jan 1995; Benjamin-Cummings; 139780805366648; 1413-1450. 
11-Wagner H., Bladt S., Zgainski E.M.; Plant Drug analysis, A thin layer chromatography atlas; 1st ed 1984; Springer; 3-540-13195-7; 117-  119. 
12-Pridham J. B.; Methods in polyphenols chemistr; 1st ed 1963; Oxford; 0080108873; 13-35, 37-48, 73-79, 131-144. 
13-Murphy J. M.; Milk thistle (Silybum marianum); The Longwood Herbal Task Force; 2000; http://www.mcp.edu/herbal/default.htm. 
14-Vladimir L.,; Method for isolation of silymarin from Silybum marianum seeds; January 15th 2008; United States Patent No. 7,318,940.  http://www.freepatentsonline.com/7318940.html. 
15-Kim N. C., Graf T. N., Sparacino C. M., Wani M. C., Wall M. E.; Complete isolation and characterization of silybins and isosilybins from milk  thistle (Silybum marianum); Org. Biomol. Chem.; 2003; (1)1684-1689. 
16-Dewick P. M; Medicinal Natural Products and Biosynthetic approach; 1st ed, 1998; John Wiley & Sons; 0-471-97477-3; 5-162,  
17-Pradhan S.C., Girish C.; Hepatoprotective herbal drug, silymarin from experimental pharmacology to clinical medicine; Ind J of Med 2006,  (124)491504. 
18-Pretsch E., Buhlmann P., Affolter C.; Structure determination of organic compounds: Table of spectral data; 1st ed 2009; Springer Verlag;  3-540-67815-8; 263-267. 
19-Silverstein R. M., Bassler  G. C., Morrill T. C.; Spectrometric identification of organic compounds; 4th ed 1981; John Wiley & Sons;  9780471029908; 88-96 . 
20-Luper Scott, N.D.; A review of plants used in the treatment of liver disease: Part 1; J of altern med rev, 1998, (3)6; 410-421. 
21-Danie S. S., Edgar J. C.; Pretreatment of milk thistle seeds to increase the silymarin yield :An alternative to petroleum ether defatting; The  New Engl J of Med, May 1 2008; 99(7): 2501-6. 
22-Wallace S. N., Jennifer R., Danielle J. C., Katherine L. V., Carl L. G., Edgar C. C., Shanmugam N.; Pressurized water versus ethanol as a  Silybum marianum extraction solvent for inhibition of low-density lipoprotein oxidation mediated by copper and J774 macrophage cells; Can  J of Physiol Pharmacol,; Sep. 2007; 85 (9):894-902 18066135. 
23-Wallace S. N., Carrier D. J., and Clausen E. C.; Batch solvent extraction of flavonolignans from Milk thistle (Silybum marianum L. Gaertner);  J of Phytochem anal.; 2005; (16)7-16. 
24-Wallace S., Clausen D. J., Robert B., Edgar C., Carl L. Griffis; HPLC-UV and 
 LC-MS-MS Characterization of Silymarin in Milk Thistle Seeds and Corresponding Products; 2003; (4)2; 37-4810. 
25-Evans W. C.; Trease and Evans Pharmacognosy; 15th ed. 2002; Elsevier; 8131200876, 9788131200872; 3-10, 137-169, 214-224. 
26-Kern Vladimirn, Walterova Daniel; Silybin and Silymarin – New effects and applications; J of Biomed papers; 2005; 149(1), 29-41. 
27-Pokharkar R. D., Takate S. B., Deshmukh R. D., Gite V. N.; Hepatoprotective activity of Launaea pinnatifida against CCl4-induced hepatic  injury in Rats; J of Pharmacol online; 2007; (2); 128-133.