Simple and Rapid HPLC Separation and Quantification of Flavonoid, Flavonolignans, and 2,3-Dehydroflavonolignans in Silymarin

. 2020 Jan 21 ; 9 (2) : . [epub] 20200121

Status PubMed-not-MEDLINE Jazyk angličtina Země Švýcarsko Médium electronic

Typ dokumentu časopisecké články

Perzistentní odkaz   https://www.medvik.cz/link/pmid31973217

Grantová podpora
18-00132S Grantová Agentura České Republiky
LTC18003, and LTC19039 Ministerstvo Školství, Mládeže a Tělovýchovy
CA16112 NutRedOx and CA16205 UNGAP European Cooperation in Science and Technology

Herbal preparations from Silybum marianum have been used since the fourth century BC in liver disease treatment and against numerous other pathologies. Consumption of silymarin containing drugs and food supplements continues to increase. Precise, fast, reliable, and complex determination of all components of silymarin preparations is paramount for assessing its pharmacological quality. We present here simple and fast HPLC-DAD and LC-MS analytical methods for the determination and quantification of all known silymarin components, including 2,3-dehydroflavonolignans that has not been achieved so far. The first method, using a common C18 column, allows baseline separation of previously inseparable silychristin A, B, isosilychristin, and silydianin. Moreover, this method allowed detection of three so far unknown silymarin components. In addition, the first analytical separation of enantiomers of 2,3-dehydrosilybin was achieved using a Lux 3μ Cellulose-4 chiral column, providing even more accurate description of silymarin composition. 2,3-Dehydroflavonolignans were isolated for the first time from silymarin using preparative chromatography on C18 and ASAHIPAK columns, and 2,3-dehydrosilychristin and 2,3-dehydrosilybin were for the first time conclusively confirmed by HPLC, MS, and NMR to be silymarin components. Using the optimized analytical methods, six various silymarin preparations were analyzed showing substantial differences in the composition.

Zobrazit více v PubMed

Martin R.J., Lauren D.R., Smith W.A., Jensen D.J., Deo B., Douglas J.A. Factors influencing silymarin content and composition in variegated thistle (Silybum marianum) N. Z. J. Crop Hortic. Sci. 2006;34:239–245. doi: 10.1080/01140671.2006.9514413. DOI

Graf T.N., Cech N.B., Polyak S.J., Oberlies N.H. A validated UHPLC-tandem mass spectrometry method for quantitative analysis of flavonolignans in milk thistle (Silybum marianum) extracts. J. Pharm. Biomed. Anal. 2016;126:26–33. doi: 10.1016/j.jpba.2016.04.028. PubMed DOI PMC

Kuki Á., Nagy L., Deák G., Nagy M., Zsuga M., Kéki S. Identification of silymarin constituents: An improved HPLC–MS method. Chromatographia. 2012;75:175–180. doi: 10.1007/s10337-011-2163-7. DOI

Wallace S., Carrier D.J., Beitle R.R., Clausen E.C., Griffis C.L. HPLC-UV and LC-MS-MS characterization of silymarin in milk thistle seeds and corresponding products. J. Nutraceutic. Funct. Med. Food. 2003;4:37–48. doi: 10.1300/J133v04n02_05. DOI

Althagafy H.S., Meza-Aviña M.E., Oberlies N.H., Croatt M.P. Mechanistic study of the biomimetic synthesis of flavonolignan diastereoisomers in milk thistle. J. Org. Chem. 2013;78:7594–7600. doi: 10.1021/jo4011377. PubMed DOI PMC

Gažák R., Svobodová A., Psotová J., Sedmera P., Přikrylová V., Walterová D., Křen V. Oxidised derivatives of silybin and their antiradical and antioxidant activity. Bioorg. Med. Chem. 2004;12:5677–5687. doi: 10.1016/j.bmc.2004.07.064. PubMed DOI

Chambers C.S., Holečková V., Petrásková L., Biedermann D., Valentová K., Buchta M., Křen V. The silymarin composition… and why does it matter? Food Res. Int. 2017;100:339–353. doi: 10.1016/j.foodres.2017.07.017. PubMed DOI

Biedermann D., Vavříková E., Cvak L., Křen V. Chemistry of silybin. Nat. Prod. Rep. 2014;31:1138–1157. doi: 10.1039/C3NP70122K. PubMed DOI

Plíšková M., Vondráček J., Křen V., Gažák R., Sedmera P., Walterová D., Psotová J., Šimánek V., Machala M. Effects of silymarin flavonolignans and synthetic silybin derivatives on estrogen and aryl hydrocarbon receptor activation. Toxicology. 2005;215:80–89. doi: 10.1016/j.tox.2005.06.020. PubMed DOI

Davis-Searles P.R., Nakanishi Y., Kim N.-C., Graf T.N., Oberlies N.H., Wani M.C., Wall M.E., Agarwal R., Kroll D.J. Milk thistle and prostate cancer: Differential effects of pure flavonolignans from Silybum marianum on antiproliferative end points in human prostate carcinoma cells. Cancer Res. 2005;65:4448–4457. doi: 10.1158/0008-5472.CAN-04-4662. PubMed DOI

Biedermann D., Buchta M., Holečková V., Sedlák D., Valentová K., Cvačka J., Bednárová L., Křenková A., Kuzma M., Škuta C., et al. Silychristin: Skeletal alterations and biological activities. J. Nat. Prod. 2016;79:3086–3092. doi: 10.1021/acs.jnatprod.6b00750. PubMed DOI

Marhol P., Bednář P., Kolářová P., Večeřa R., Ulrichová J., Tesařová E., Vavříková E., Kuzma M., Křen V. Pharmacokinetics of pure silybin diastereoisomers and identification of their metabolites in rat plasma. J. Funct. Food. 2015;14:570–580. doi: 10.1016/j.jff.2015.02.031. DOI

Filippopoulou K., Papaevgeniou N., Lefaki M., Paraskevopoulou A., Biedermann D., Křen V., Chondrogianni N. 2,3-Dehydrosilybin A/B as a pro-longevity and anti-aggregation compound. Free Radic. Biol. Med. 2017;103:256–267. doi: 10.1016/j.freeradbiomed.2016.12.042. PubMed DOI

Rajnochová Svobodová A., Gabrielová E., Ulrichová J., Zálešák B., Biedermann D., Vostálová J. A pilot study of the UVA-photoprotective potential of dehydrosilybin, isosilybin, silychristin, and silydianin on human dermal fibroblasts. Arch. Dermatol. Res. 2019;311:477–490. doi: 10.1007/s00403-019-01928-7. PubMed DOI

Kosina P., Paloncýová M., Rajnochová Svobodová A., Zálešák B., Biedermann D., Ulrichová J., Vostálová J. Dermal delivery of selected polyphenols from Silybum marianum. Theoretical and experimental study. Molecules. 2018;24:61. doi: 10.3390/molecules24010061. PubMed DOI PMC

Vostálová J., Tinková E., Biedermann D., Kosina P., Ulrichová J., Rajnochová Svobodová A. Skin protective activity of silymarin and its flavonolignans. Molecules. 2019;24:1022. doi: 10.3390/molecules24061022. PubMed DOI PMC

Tilley C., Deep G., Agarwal C., Wempe M.F., Biedermann D., Valentová K., Kren V., Agarwal R. Silibinin and its 2,3-dehydro-derivative inhibit basal cell carcinoma growth via suppression of mitogenic signaling and transcription factors activation. Mol. Carcinog. 2016;55:3–14. doi: 10.1002/mc.22253. PubMed DOI PMC

Fenclová M., Nováková A., Viktorová J., Jonatová P., Džuman Z., Ruml T., Křen V., Hajšlová J., Vítek L., Stránská-Zachariášová M. Poor chemical and microbiological quality of the commercial milk thistle-based dietary supplements may account for their reported unsatisfactory and non-reproducible clinical outcomes. Sci. Rep. 2019;9:11118. doi: 10.1038/s41598-019-47250-0. PubMed DOI PMC

Csupor D., Csorba A., Hohmann J. Recent advances in the analysis of flavonolignans of Silybum marianum. J. Pharm. Biomed. Anal. 2016;130:301–317. doi: 10.1016/j.jpba.2016.05.034. PubMed DOI

Karkanis A., Bilalis D., Efthimiadou A. Cultivation of milk thistle (Silybum marianum L. Gaertn.), a medicinal weed. Ind. Crop. Prod. 2011;34:825–830. doi: 10.1016/j.indcrop.2011.03.027. DOI

Džubák P., Hajdúch M., Gažák R., Svobodová A., Psotová J., Walterová D., Sedmera P., Křen V. New derivatives of silybin and 2,3-dehydrosilybin and their cytotoxic and P-glycoprotein modulatory activity. Bioorg. Med. Chem. 2006;14:3793–3810. doi: 10.1016/j.bmc.2006.01.035. PubMed DOI

Biedermann D., Moravcová V., Valentová K., Kuzma M., Petrásková L., Císařová I., Křen V. Oxidation of flavonolignan silydianin to unexpected lactone-acid derivative. Phytochem. Lett. 2019;30:14–20. doi: 10.1016/j.phytol.2019.01.006. DOI

Křenek K., Marhol P., Peikerová Ž., Křen V., Biedermann D. Preparatory separation of the silymarin flavonolignans by Sephadex LH-20 gel. Food Res. Int. 2014;65:115–120. doi: 10.1016/j.foodres.2014.02.001. DOI

Pyszková M., Biler M., Biedermann D., Valentová K., Kuzma M., Vrba J., Ulrichová J., Sokolová R., Mojovic M., Popovic-Bijelic A., et al. Flavonolignan 2,3-dehydroderivatives: Preparation, antiradical and cytoprotective activity. Free Radic. Biol. Med. 2016;90:114–125. doi: 10.1016/j.freeradbiomed.2015.11.014. PubMed DOI

Novotná M., Gažák R., Biedermann D., Di Meo F., Marhol P., Kuzma M., Bednárová L., Fuksová K., Trouillas P., Křen V. cis–trans Isomerization of silybins A and B. Beilstein J. Org. Chem. 2014;10:1047–1063. doi: 10.3762/bjoc.10.105. PubMed DOI PMC

Liu H., Du Z., Yuan Q. A novel rapid method for simultaneous determination of eight active compounds in silymarin using a reversed-phase UPLC-UV detector. J. Chromatogr. B. 2009;877:4159–4163. doi: 10.1016/j.jchromb.2009.11.001. PubMed DOI

AbouZid S.F., Chen S.N., Pauli G.F. Silymarin content in Silybum marianum populations growing in Egypt. Ind. Crop. Prod. 2016;83:729–737. doi: 10.1016/j.indcrop.2015.12.012. PubMed DOI PMC

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. doi: 10.1039/b300099k. PubMed DOI

Cai X., Li D., Qiao J., Lian H., Wang S. Determination of silymarin flavonoids by HPLC and LC-MS and investigation of extraction rate of silymarin in Silybum marianum fruits by boiling water. Asian J. Chem. 2009;21:63–74.

Keshavarz Afshar R., Chaichi M.R., Ansari Jovini M., Jahanzad E., Hashemi M. Accumulation of silymarin in milk thistle seeds under drought stress. Planta. 2015;242:539–543. doi: 10.1007/s00425-015-2265-9. PubMed DOI

Poppe L., Petersen M. Variation in the flavonolignan composition of fruits from different Silybum marianum chemotypes and suspension cultures derived therefrom. Phytochemistry. 2016;131:68–75. doi: 10.1016/j.phytochem.2016.09.003. PubMed DOI

Bilia A.R., Salvini D., Mazzi G., Vincieri F.F. Characterization of calendula flower, milk-thistle fruit, and passion flower tinctures by HPLC-DAD and HPLC-MS. Chromatographia. 2000;53:210–215. doi: 10.1007/BF02491573. DOI

Gažák R., Sedmera P., Marzorati M., Riva S., Křen V. Laccase-mediated dimerization of the flavonolignan silybin. J. Mol. Catal. B Enzym. 2008;50:87–92. doi: 10.1016/j.molcatb.2007.09.005. DOI

Nejnovějších 20 citací...

Zobrazit více v
Medvik | PubMed

Effects of catechins, resveratrol, silymarin components and some of their conjugates on xanthine oxidase-catalyzed xanthine and 6-mercaptopurine oxidation

. 2025 Mar 30 ; 105 (5) : 2765-2776. [epub] 20241128

Flavonolignans silybin, silychristin and 2,3-dehydrosilybin showed differential cytoprotective, antioxidant and anti-apoptotic effects on splenocytes from Balb/c mice

. 2025 Feb 15 ; 15 (1) : 5631. [epub] 20250215

Mutual Interactions of Silymarin and Colon Microbiota in Healthy Young and Healthy Elder Subjects

. 2024 Nov ; 68 (22) : e2400500. [epub] 20241030

Sulfated Phenolic Substances: Preparation and Optimized HPLC Analysis

. 2022 May 20 ; 23 (10) : . [epub] 20220520

Silymarin Dehydroflavonolignans Chelate Zinc and Partially Inhibit Alcohol Dehydrogenase

. 2021 Nov 25 ; 13 (12) : . [epub] 20211125

Chirality Matters: Biological Activity of Optically Pure Silybin and Its Congeners

. 2021 Jul 23 ; 22 (15) : . [epub] 20210723

Metabolism of 2,3-Dehydrosilybin A and 2,3-Dehydrosilybin B: A Study with Human Hepatocytes and Recombinant UDP-Glucuronosyltransferases and Sulfotransferases

. 2021 Jun 14 ; 10 (6) : . [epub] 20210614

Dehydroflavonolignans from Silymarin Potentiate Transition Metal Toxicity In Vitro but Are Protective for Isolated Erythrocytes Ex Vivo

. 2021 Apr 27 ; 10 (5) : . [epub] 20210427

Identification of Human Sulfotransferases Active towards Silymarin Flavonolignans and Taxifolin

. 2020 Aug 12 ; 10 (8) : . [epub] 20200812

Multidrug Resistance Modulation Activity of Silybin Derivatives and Their Anti-inflammatory Potential

. 2020 May 25 ; 9 (5) : . [epub] 20200525

Dual SMO/BRAF Inhibition by Flavonolignans from Silybum marianum †

. 2020 May 05 ; 9 (5) : . [epub] 20200505

Biotransformation of Silymarin Flavonolignans by Human Fecal Microbiota

. 2020 Jan 09 ; 10 (1) : . [epub] 20200109

Najít záznam

Citační ukazatele

Nahrávání dat ...

Možnosti archivace

Nahrávání dat ...