بررسی استفاده از آب در استخراج سیلی‌مارین از دانه‌های گیاه خارمریم (Silybum marianum L. Gaertn)

الموضوعات :

طاهره حسنلو ¹ , روشنک سپهری فر ²

1 - بخش فیزیولوژی، مؤسسه تحقیقات بیوتکنولوژی کشاورزی، ابتدای جاده ماهدشت، کرج
2 - بخش فیزیولوژی، مؤسسه تحقیقات بیوتکنولوژی کشاورزی، ابتدای جاده ماهدشت، کرج

تاريخ الإرسال : 19 السبت , جمادى الثانية, 1427 تاريخ التأكيد : 05 السبت , ذو الحجة, 1428 تاريخ الإصدار : 13 الأربعاء , صفر, 1429

الکلمات المفتاحية: حلال, سیلیمارین, خار مریم,

ملخص المقالة :

سیلی مارین ترکیبی دارویی است که از دانه‌های گیاه دارویی خار مریم (Silybum marianum (L.) Gaertn) استخراج و در درمان بیماری های کبدی, دیابت نوع П و کلسترول بالا استفاده می شود. به منظور استخراج سیلی مارین از حلال های آلی همچون متانول, اتانول, استونیتریل و استون استفاده می‌گردد. در این تحقیق به منظور مقایسه استفاده از حلال آب با توجه به ارزانی, سهولت و بی خطر بودن به عنوان یک حلال مناسب پیشنهاد شده است. لذا با روش‌های مختلف, مقدار و نوع ترکیب‌های فلاونولیگنانی عصاره آبی استخراج شده از دانه‌های گیاه خار مریم توسط روش کروماتوگرافی مایع با کارایی بالا اندازه‌گیری شد. نتایج نشان داد که بیشترین مقدار تاکسی فولین, سیلی کریستین و سیلی دیانین استخراج شده (به ترتیب 6/0، 9/0 و 3/2 میلی گرم در گرم ماده خشک) با استفاده از روش سوکسله با آب بعد از انجام یک مرحله چربی‌زدایی می‌باشد. بیشترین مقدار سیلی بین و ایزوسیلی بین استخراج شده (به ترتیب 24/0 , 15/0 میلی گرم در گرم ماده خشک) با استفاده از روش سوکسله با آب بدون انجام چربی زدایی قبل از استخراج می‌باشد. در مقایسه با متانول به عنوان حلال مقدار ماده استخراجی با آب بسیار پایین تر بود. به منظور افزایش بازده, با تغییر روش وافزایش زمان استخراج می‌توان در آینده به نتایج بهتری دست یافت.

المصادر:

حسنلو، ط.، خاوری نژاد، ر.ع.، مجیدی، ا. و ضیایی س. ع. (1383) مطالعه و تعیین فلاونولیگنان ها در میوه‌های گیاه خارمریم جمع آوری شده از نقاط مختلف ایران به روش‌های اسپکتروفتومتری TLC و HPLC. فصلنامه گیاهان دارویی، ویژه نامه خارمریم، سال چهارم، صفحات 25-32.

ضیایی، س. ع.، حسینی، ح. ف.، رجبیان، ط.، پور حسینی، ل.، نقدی بادی، ح. و رضازاده، ش. (1383) بررسی اثر حلال­های مختلف در  استخراج  سیلی­مارین از بذر گیاه خارمریم. فصلنامه گیاهان دارویی، ویژه نامه خارمریم، سال چهارم، صفحات 7-12.

ولاگ، ژ. و استودولا، ژ.، ترجمه ساعد زمان، س.(1376) گیاهان دارویی، انتشارات دانشگاه تهران، صفحه 309.

 

 

Alikaridis, F., D. Papadakis, K. Pantelia and T. Kephalas (2000) Flavonolignan  production from Silybum marianum transformed and untransformed root culture. Fitotrapia, 71: 379-384.

Alvarezbarreto, J., S. N. Wallace, D. J. Carrier and E. C. Clausene (2003) Extraction of nutraceuticals from milk thistle. I. Hot water extraction. Applied Biochemistry and Biotechnology, 105: 881-889.

Basile, A., M. Jimenez-Carmona and A. Clifford (1998) Extraction of rosemary by superheated water. Journal of Agricultural and Food Chemistry, 46: 5205-5209.

Benthin B., H. Danz and M. Hamburger (1999) Pressurized liquid extraction of medicinal plants. Journal of Chromatography A, 837: 211-219.

Bosisio, E., C. Benelli and O. Pirola (1992) Effect of the flavonolignans of Silybum marianum L. Gaertn on lipid peroxidation in rat liver microsomes and freshly isolated hepatocytes. Pharmacology Research, 25: 147-154.

Bourgand, F., A. Gravot, S. Milesi and E. Gontier (2001) Production of plant secondary metabolites: a historical perspective. Plant Science, 161: 839-851.

Brenda, W.S. (2000) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiology, 126: 485-493.

Briskin, D.P. (2000) Medicinal plants and phytomedicines. linking plant biochemistry and physiology to human health. Plant Physiology, 124: 507-514.

Cacho, M., M. Moran, P. Corchete and J. Fernandez-Tarrago (1999) Influence of medium composition on the accumulation of flavonolignans in cultured cells of Silybum marianum (L.)Gaertn. Plant Science, 144: 63-68.

 

 

 

Ding, T.M., S.J. Tian Z.X. Zhang, D.Z. Gu, Y.F. Chen, Y.H. Shi and Z.P. Sun (2001) Determination of active component in silymarin by RP-LC and LC/MS. Journal of Phamaceutical and Biochemical Analysis, 26: 155-161.

Duan, L., D. J. Carrier and E. C. Clausen (2004) Silymarin extraction from milk thistle using hot water. Applied Biochemistry and Biotechnology, 114: 559-568.

Hasanloo, T., R. A. Khavari-Nejad, E. Majidi and M. R. Shams- Ardakani (2005) Analysis of flavonolignans in dried fruits of Silybum marianum (L.) Gaertn from Iran. Pakistan Journal of Biological Science, 8: 1778-1782.

Kahol, A., K. Singh, S. Tandon and S. Kumar (2001) Process for isolation of hepaprotective agent silymarin from the seeds of the plant silybum marianum. Indian Patent Number: 06309678.

Katiyar, S. K. (2002) Treatment of silymarin, a plant flavonoid, prevents ultraviolet light- induced immune suppression and oxidative stress in mouse skin. International Journal of Oncology, 21: 1213-1222.

Kren, V., J. Ulrichova, P. Kosina, D. Stevenson, P. Sedmera, V. Prikrylova, P. Halada and V. Simanek (2000) Chemoenzymatic preparation of silybin β- glucoronides and their biological evaluation. Drag Metobolism and Disposition, 28: 1513-1517.

Kubatova A., D. Miller and S. Hawthorne (2001) Selective extraction of oxygenated natural products with subcritical water. 10^th International symposium and exhibit on supercritical fluid chromatography, extraction and processing. August 19-22, 2001. Myrtle Beach, SC, USA. E-05.

Li, W., K. Koike, Y. Asada, T. Yoshikawa and T. Nikaido (2005) Rosmarinic acid production by coleus forskohlii hairy root cultures. Plant Cell, Tissue and Organ Cultur, 80: 151-155.

Minakhmetov, R. A., L. A. Onuchak, V. A. Kurkin, E. V. Avdeeva and A. V. Volotsueva (2001) Analysis of flavonoids in silybum marianum fruit by HPLC. Chemistry of Natural Compounds, 3: 318-321.

Narayana, K.R., R. Sripal, M. R. Chaluvadi and D. R. Krishan, D.R. (2001) Bioflavonoids classification, pharmacological, biochemical effects and therapeutic potential. Indian Journal of Pharmacology, 33:2-6.

Quaglia, M.G., E. Bossu, E. Donati, G. Mazzanti and A. Brandt (1999) Determination of silymarin in the extract from the dried Silybum marianum fruits by high performance liquid chromatography and capillary electrophoresis. Journal of Pharmaceutical and Biomedical Analysis, 19: 435-442.

Samuelsson, G. (1999) Drugs of natural origin. 4th revised edition. Swedish pharmaceutical press, Stockholm, Sweden. 226-233.

Sánchez-Sampedro, A., H.K. Kim, H.Y. Choi, R. Verpoorte and P. Corchete (2007) Metabolomic alterations in elicitor treated Silybum marianum suspension cultures monitored by nuclear magnetic resonance spectroscopy. J Biotechnol., 130: 133-142.

Schnfeld, J.V., B. Weisbrod and M. K. Muller (1997) Silibinin, a plant extract with antioxidant and membrane stabilizing properties protects exocrine pancreas from cyclosporine A toxicity. Cellular and Molecular Life Sciences, 53:917-920.

Sonnenbichler, J., F. Scaleram, I. Sonnenbichler and R. Weyhenmeyer (1999) Stimulatory effects of silibinin and silichristin form the Milk thistle silybum marianum on kidney cells. The Journal of Pharmacology and Experimental Therapeutics, 290: 1375-1383.

Titel, G. and H. Wagner (1977) High-performance liquid chromatographic seperation of silymarins and their determination in a raw extract of silybum marianum. Journal of Chromatography, 135: 499-501.

Venkataramanan, R. (2000) Milke thistle, a herbal supplement, decreases the activity of CYP3A4 and uridine diphosphoglucuronosyl transferase in human hepatocyte cultures. Drug Metabolism and Dispositions, 28: 1270-1273.

Wagner, H., P. Diesel and M. Seites (1974) The chemistry and analysis of silymarin from Silybum marianum. Arzheimittle Forschung, 24: 466-71.

Wagner, H., L. Horhammer and M. Seitz (1968) Chemical evaluation of a silymarin-contaning   flavonoid concentrate from Silybum marianum. Arzheimittel Forschung, 18: 696-8.

Wallace, S. N., D. J. Carrier and E. C. Clausen (2003) Extraction of nutraceuticals from milk thistle: part II. Extraction with organic solvents. Applied Biochemistry Biotechnology, 105: 891-903.

Wallace, S. N., D. J. Carrier and E. C. Clausen (2005) Batch solvent extraction of flavanolignans from milk thistle (Silybum marianum L. Gaertn). Phytochemical Analysis, 16: 7-16

حسنلو، ط.، خاوری نژاد، ر.ع.، مجیدی، ا. و ضیایی س. ع. (1383) مطالعه و تعیین فلاونولیگنان ها در میوه‌های گیاه خارمریم جمع آوری شده از نقاط مختلف ایران به روش‌های اسپکتروفتومتری TLC و HPLC. فصلنامه گیاهان دارویی، ویژه نامه خارمریم، سال چهارم، صفحات 25-32.

ضیایی، س. ع.، حسینی، ح. ف.، رجبیان، ط.، پور حسینی، ل.، نقدی بادی، ح. و رضازاده، ش. (1383) بررسی اثر حلال­های مختلف در  استخراج  سیلی­مارین از بذر گیاه خارمریم. فصلنامه گیاهان دارویی، ویژه نامه خارمریم، سال چهارم، صفحات 7-12.

ولاگ، ژ. و استودولا، ژ.، ترجمه ساعد زمان، س.(1376) گیاهان دارویی، انتشارات دانشگاه تهران، صفحه 309.

 

 

Alikaridis, F., D. Papadakis, K. Pantelia and T. Kephalas (2000) Flavonolignan  production from Silybum marianum transformed and untransformed root culture. Fitotrapia, 71: 379-384.

Alvarezbarreto, J., S. N. Wallace, D. J. Carrier and E. C. Clausene (2003) Extraction of nutraceuticals from milk thistle. I. Hot water extraction. Applied Biochemistry and Biotechnology, 105: 881-889.

Basile, A., M. Jimenez-Carmona and A. Clifford (1998) Extraction of rosemary by superheated water. Journal of Agricultural and Food Chemistry, 46: 5205-5209.

Benthin B., H. Danz and M. Hamburger (1999) Pressurized liquid extraction of medicinal plants. Journal of Chromatography A, 837: 211-219.

Bosisio, E., C. Benelli and O. Pirola (1992) Effect of the flavonolignans of Silybum marianum L. Gaertn on lipid peroxidation in rat liver microsomes and freshly isolated hepatocytes. Pharmacology Research, 25: 147-154.

Bourgand, F., A. Gravot, S. Milesi and E. Gontier (2001) Production of plant secondary metabolites: a historical perspective. Plant Science, 161: 839-851.

Brenda, W.S. (2000) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiology, 126: 485-493.

Briskin, D.P. (2000) Medicinal plants and phytomedicines. linking plant biochemistry and physiology to human health. Plant Physiology, 124: 507-514.

Cacho, M., M. Moran, P. Corchete and J. Fernandez-Tarrago (1999) Influence of medium composition on the accumulation of flavonolignans in cultured cells of Silybum marianum (L.)Gaertn. Plant Science, 144: 63-68.

 

 

 

Ding, T.M., S.J. Tian Z.X. Zhang, D.Z. Gu, Y.F. Chen, Y.H. Shi and Z.P. Sun (2001) Determination of active component in silymarin by RP-LC and LC/MS. Journal of Phamaceutical and Biochemical Analysis, 26: 155-161.

Duan, L., D. J. Carrier and E. C. Clausen (2004) Silymarin extraction from milk thistle using hot water. Applied Biochemistry and Biotechnology, 114: 559-568.

Hasanloo, T., R. A. Khavari-Nejad, E. Majidi and M. R. Shams- Ardakani (2005) Analysis of flavonolignans in dried fruits of Silybum marianum (L.) Gaertn from Iran. Pakistan Journal of Biological Science, 8: 1778-1782.

Kahol, A., K. Singh, S. Tandon and S. Kumar (2001) Process for isolation of hepaprotective agent silymarin from the seeds of the plant silybum marianum. Indian Patent Number: 06309678.

Katiyar, S. K. (2002) Treatment of silymarin, a plant flavonoid, prevents ultraviolet light- induced immune suppression and oxidative stress in mouse skin. International Journal of Oncology, 21: 1213-1222.

Kren, V., J. Ulrichova, P. Kosina, D. Stevenson, P. Sedmera, V. Prikrylova, P. Halada and V. Simanek (2000) Chemoenzymatic preparation of silybin β- glucoronides and their biological evaluation. Drag Metobolism and Disposition, 28: 1513-1517.

Kubatova A., D. Miller and S. Hawthorne (2001) Selective extraction of oxygenated natural products with subcritical water. 10^th International symposium and exhibit on supercritical fluid chromatography, extraction and processing. August 19-22, 2001. Myrtle Beach, SC, USA. E-05.

Li, W., K. Koike, Y. Asada, T. Yoshikawa and T. Nikaido (2005) Rosmarinic acid production by coleus forskohlii hairy root cultures. Plant Cell, Tissue and Organ Cultur, 80: 151-155.

Minakhmetov, R. A., L. A. Onuchak, V. A. Kurkin, E. V. Avdeeva and A. V. Volotsueva (2001) Analysis of flavonoids in silybum marianum fruit by HPLC. Chemistry of Natural Compounds, 3: 318-321.

Narayana, K.R., R. Sripal, M. R. Chaluvadi and D. R. Krishan, D.R. (2001) Bioflavonoids classification, pharmacological, biochemical effects and therapeutic potential. Indian Journal of Pharmacology, 33:2-6.

Quaglia, M.G., E. Bossu, E. Donati, G. Mazzanti and A. Brandt (1999) Determination of silymarin in the extract from the dried Silybum marianum fruits by high performance liquid chromatography and capillary electrophoresis. Journal of Pharmaceutical and Biomedical Analysis, 19: 435-442.

Samuelsson, G. (1999) Drugs of natural origin. 4th revised edition. Swedish pharmaceutical press, Stockholm, Sweden. 226-233.

Sánchez-Sampedro, A., H.K. Kim, H.Y. Choi, R. Verpoorte and P. Corchete (2007) Metabolomic alterations in elicitor treated Silybum marianum suspension cultures monitored by nuclear magnetic resonance spectroscopy. J Biotechnol., 130: 133-142.

Schnfeld, J.V., B. Weisbrod and M. K. Muller (1997) Silibinin, a plant extract with antioxidant and membrane stabilizing properties protects exocrine pancreas from cyclosporine A toxicity. Cellular and Molecular Life Sciences, 53:917-920.

Sonnenbichler, J., F. Scaleram, I. Sonnenbichler and R. Weyhenmeyer (1999) Stimulatory effects of silibinin and silichristin form the Milk thistle silybum marianum on kidney cells. The Journal of Pharmacology and Experimental Therapeutics, 290: 1375-1383.

Titel, G. and H. Wagner (1977) High-performance liquid chromatographic seperation of silymarins and their determination in a raw extract of silybum marianum. Journal of Chromatography, 135: 499-501.

Venkataramanan, R. (2000) Milke thistle, a herbal supplement, decreases the activity of CYP3A4 and uridine diphosphoglucuronosyl transferase in human hepatocyte cultures. Drug Metabolism and Dispositions, 28: 1270-1273.

Wagner, H., P. Diesel and M. Seites (1974) The chemistry and analysis of silymarin from Silybum marianum. Arzheimittle Forschung, 24: 466-71.

Wagner, H., L. Horhammer and M. Seitz (1968) Chemical evaluation of a silymarin-contaning   flavonoid concentrate from Silybum marianum. Arzheimittel Forschung, 18: 696-8.

Wallace, S. N., D. J. Carrier and E. C. Clausen (2003) Extraction of nutraceuticals from milk thistle: part II. Extraction with organic solvents. Applied Biochemistry Biotechnology, 105: 891-903.

Wallace, S. N., D. J. Carrier and E. C. Clausen (2005) Batch solvent extraction of flavanolignans from milk thistle (Silybum marianum L. Gaertn). Phytochemical Analysis, 16: 7-16

 

حسنلو، ط.، خاوری نژاد، ر.ع.، مجیدی، ا. و ضیایی س. ع. (1383) مطالعه و تعیین فلاونولیگنان ها در میوه‌های گیاه خارمریم جمع آوری شده از نقاط مختلف ایران به روش‌های اسپکتروفتومتری TLC و HPLC. فصلنامه گیاهان دارویی، ویژه نامه خارمریم، سال چهارم، صفحات 25-32.

ضیایی، س. ع.، حسینی، ح. ف.، رجبیان، ط.، پور حسینی، ل.، نقدی بادی، ح. و رضازاده، ش. (1383) بررسی اثر حلال­های مختلف در  استخراج  سیلی­مارین از بذر گیاه خارمریم. فصلنامه گیاهان دارویی، ویژه نامه خارمریم، سال چهارم، صفحات 7-12.

ولاگ، ژ. و استودولا، ژ.، ترجمه ساعد زمان، س.(1376) گیاهان دارویی، انتشارات دانشگاه تهران، صفحه 309.

 

 

Alikaridis, F., D. Papadakis, K. Pantelia and T. Kephalas (2000) Flavonolignan  production from Silybum marianum transformed and untransformed root culture. Fitotrapia, 71: 379-384.

Alvarezbarreto, J., S. N. Wallace, D. J. Carrier and E. C. Clausene (2003) Extraction of nutraceuticals from milk thistle. I. Hot water extraction. Applied Biochemistry and Biotechnology, 105: 881-889.

Basile, A., M. Jimenez-Carmona and A. Clifford (1998) Extraction of rosemary by superheated water. Journal of Agricultural and Food Chemistry, 46: 5205-5209.

Benthin B., H. Danz and M. Hamburger (1999) Pressurized liquid extraction of medicinal plants. Journal of Chromatography A, 837: 211-219.

Bosisio, E., C. Benelli and O. Pirola (1992) Effect of the flavonolignans of Silybum marianum L. Gaertn on lipid peroxidation in rat liver microsomes and freshly isolated hepatocytes. Pharmacology Research, 25: 147-154.

Bourgand, F., A. Gravot, S. Milesi and E. Gontier (2001) Production of plant secondary metabolites: a historical perspective. Plant Science, 161: 839-851.

Brenda, W.S. (2000) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiology, 126: 485-493.

Briskin, D.P. (2000) Medicinal plants and phytomedicines. linking plant biochemistry and physiology to human health. Plant Physiology, 124: 507-514.

Cacho, M., M. Moran, P. Corchete and J. Fernandez-Tarrago (1999) Influence of medium composition on the accumulation of flavonolignans in cultured cells of Silybum marianum (L.)Gaertn. Plant Science, 144: 63-68.

 

 

 

Ding, T.M., S.J. Tian Z.X. Zhang, D.Z. Gu, Y.F. Chen, Y.H. Shi and Z.P. Sun (2001) Determination of active component in silymarin by RP-LC and LC/MS. Journal of Phamaceutical and Biochemical Analysis, 26: 155-161.

Duan, L., D. J. Carrier and E. C. Clausen (2004) Silymarin extraction from milk thistle using hot water. Applied Biochemistry and Biotechnology, 114: 559-568.

Hasanloo, T., R. A. Khavari-Nejad, E. Majidi and M. R. Shams- Ardakani (2005) Analysis of flavonolignans in dried fruits of Silybum marianum (L.) Gaertn from Iran. Pakistan Journal of Biological Science, 8: 1778-1782.

Kahol, A., K. Singh, S. Tandon and S. Kumar (2001) Process for isolation of hepaprotective agent silymarin from the seeds of the plant silybum marianum. Indian Patent Number: 06309678.

Katiyar, S. K. (2002) Treatment of silymarin, a plant flavonoid, prevents ultraviolet light- induced immune suppression and oxidative stress in mouse skin. International Journal of Oncology, 21: 1213-1222.

Kren, V., J. Ulrichova, P. Kosina, D. Stevenson, P. Sedmera, V. Prikrylova, P. Halada and V. Simanek (2000) Chemoenzymatic preparation of silybin β- glucoronides and their biological evaluation. Drag Metobolism and Disposition, 28: 1513-1517.

Kubatova A., D. Miller and S. Hawthorne (2001) Selective extraction of oxygenated natural products with subcritical water. 10^th International symposium and exhibit on supercritical fluid chromatography, extraction and processing. August 19-22, 2001. Myrtle Beach, SC, USA. E-05.

Li, W., K. Koike, Y. Asada, T. Yoshikawa and T. Nikaido (2005) Rosmarinic acid production by coleus forskohlii hairy root cultures. Plant Cell, Tissue and Organ Cultur, 80: 151-155.

Minakhmetov, R. A., L. A. Onuchak, V. A. Kurkin, E. V. Avdeeva and A. V. Volotsueva (2001) Analysis of flavonoids in silybum marianum fruit by HPLC. Chemistry of Natural Compounds, 3: 318-321.

Narayana, K.R., R. Sripal, M. R. Chaluvadi and D. R. Krishan, D.R. (2001) Bioflavonoids classification, pharmacological, biochemical effects and therapeutic potential. Indian Journal of Pharmacology, 33:2-6.

Quaglia, M.G., E. Bossu, E. Donati, G. Mazzanti and A. Brandt (1999) Determination of silymarin in the extract from the dried Silybum marianum fruits by high performance liquid chromatography and capillary electrophoresis. Journal of Pharmaceutical and Biomedical Analysis, 19: 435-442.

Samuelsson, G. (1999) Drugs of natural origin. 4th revised edition. Swedish pharmaceutical press, Stockholm, Sweden. 226-233.

Sánchez-Sampedro, A., H.K. Kim, H.Y. Choi, R. Verpoorte and P. Corchete (2007) Metabolomic alterations in elicitor treated Silybum marianum suspension cultures monitored by nuclear magnetic resonance spectroscopy. J Biotechnol., 130: 133-142.

Schnfeld, J.V., B. Weisbrod and M. K. Muller (1997) Silibinin, a plant extract with antioxidant and membrane stabilizing properties protects exocrine pancreas from cyclosporine A toxicity. Cellular and Molecular Life Sciences, 53:917-920.

Sonnenbichler, J., F. Scaleram, I. Sonnenbichler and R. Weyhenmeyer (1999) Stimulatory effects of silibinin and silichristin form the Milk thistle silybum marianum on kidney cells. The Journal of Pharmacology and Experimental Therapeutics, 290: 1375-1383.

Titel, G. and H. Wagner (1977) High-performance liquid chromatographic seperation of silymarins and their determination in a raw extract of silybum marianum. Journal of Chromatography, 135: 499-501.

Venkataramanan, R. (2000) Milke thistle, a herbal supplement, decreases the activity of CYP3A4 and uridine diphosphoglucuronosyl transferase in human hepatocyte cultures. Drug Metabolism and Dispositions, 28: 1270-1273.

Wagner, H., P. Diesel and M. Seites (1974) The chemistry and analysis of silymarin from Silybum marianum. Arzheimittle Forschung, 24: 466-71.

Wagner, H., L. Horhammer and M. Seitz (1968) Chemical evaluation of a silymarin-contaning   flavonoid concentrate from Silybum marianum. Arzheimittel Forschung, 18: 696-8.

Wallace, S. N., D. J. Carrier and E. C. Clausen (2003) Extraction of nutraceuticals from milk thistle: part II. Extraction with organic solvents. Applied Biochemistry Biotechnology, 105: 891-903.

Wallace, S. N., D. J. Carrier and E. C. Clausen (2005) Batch solvent extraction of flavanolignans from milk thistle (Silybum marianum L. Gaertn). Phytochemical Analysis, 16: 7-16