Tracking and quantifying the major silymarin flavonolignans in the fruit extracts of milk thistle (Silybum marianum L.) using a validated HPTLC-MS method
Mahgoub, Y., El-Hawiet, A., El Sebakhy, N., Darwish, F., Shawky, E. (2025). Tracking and quantifying the major silymarin flavonolignans in the fruit extracts of milk thistle (Silybum marianum L.) using a validated HPTLC-MS method. EKB Journal Management System, 2(1), 17-23. doi: 10.21608/japs.2024.290405.1027
Yasmin Adel Mahgoub; Amr Moustafa El-Hawiet; Nadia Abdel Aziz El Sebakhy; Fikria Ali Darwish; Eman Shawky. "Tracking and quantifying the major silymarin flavonolignans in the fruit extracts of milk thistle (Silybum marianum L.) using a validated HPTLC-MS method". EKB Journal Management System, 2, 1, 2025, 17-23. doi: 10.21608/japs.2024.290405.1027
Mahgoub, Y., El-Hawiet, A., El Sebakhy, N., Darwish, F., Shawky, E. (2025). 'Tracking and quantifying the major silymarin flavonolignans in the fruit extracts of milk thistle (Silybum marianum L.) using a validated HPTLC-MS method', EKB Journal Management System, 2(1), pp. 17-23. doi: 10.21608/japs.2024.290405.1027
Mahgoub, Y., El-Hawiet, A., El Sebakhy, N., Darwish, F., Shawky, E. Tracking and quantifying the major silymarin flavonolignans in the fruit extracts of milk thistle (Silybum marianum L.) using a validated HPTLC-MS method. EKB Journal Management System, 2025; 2(1): 17-23. doi: 10.21608/japs.2024.290405.1027

Tracking and quantifying the major silymarin flavonolignans in the fruit extracts of milk thistle (Silybum marianum L.) using a validated HPTLC-MS method

Journal of Advanced Pharmaceutical Sciences
Volume 2, Issue 1, January 2025, Page 17-23  XML PDF (617.85 K)
Document Type: Short Communication
DOI: 10.21608/japs.2024.290405.1027
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Authors
Yasmin Adel Mahgoub email ; Amr Moustafa El-Hawiet; Nadia Abdel Aziz El Sebakhy; Fikria Ali Darwish; Eman Shawky
Department of Pharmacognosy, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt
Abstract
High performance thin layer chromatography hyphenated with mass spectrometry (HPTLC-MS) was implemented for the first time for the simultaneous tracking and quantitation of the major flavonolignans in fruit extracts of milk thistle (Silybum marianum L.) varieties; marianum and albiflora.  A mobile phase of chloroform: acetone: formic acid (110: 16.5: 8.5 v/v/v) was utilized for the development of the tracked flavonolignans followed by their visualization under UV 366 nm after NP/PEG derivatization. The tracked flavonolignans were identified, using mass analysis of the developed flavonolignans directly from the developed plate, and quantified. The developed method's linearity, accuracy, and precision were all confirmed in accordance with ICH requirements. Linear calibrations for the quantified flavonolignans were obtained with a correlation coefficient > 0.98. Precision was confirmed with a relative standard deviation ranging between 1.34 and 1.83%.  In addition, acceptable accuracy was obtained with recovery exceeding 97%, as well as appropriate limit of detection (LOD) and limit of quantitation (LOQ).
 
Keywords
HPTLC-MS; Silybum varieties; Flavonolignans
References
  1. Boulos L. Flora of Egypt, Volume three (Verbenaceae - Compositae). Al Hadara Publishing; Cairo, Egypt; 2002. 157–159 p.
  2. Mashaly IA, El shahaby OA EAY. Floristic features of the canal bank habitats, Egypt. Journal of environmental sciences. 2010;39(4):483–501.
  3. Marceddu R, Dinolfo L, Carrubba A, Sarno M, Di Miceli G. Milk Thistle (Silybum Marianum L.) as a Novel Multipurpose Crop for Agriculture in Marginal Environments: A Review. Vol. 12, Agronomy. MDPI; 2022.
  4. Wang X, Zhang Z, Wu SC. Health Benefits of Silybum marianum: Phytochemistry, Pharmacology, and Applications. J Agric Food Chem. 2020 Oct 21;68(42):11644–64.
  5. Song K, Li M, Yang Y, Zhang Z, Zhu Q, Liu J, et al. Natural flavonolignans as potential therapeutic agents against common diseases. Vol. 74, Journal of Pharmacy and Pharmacology. Oxford University Press; 2022. p. 337–50.
  6. Yin F, Liu J, Ji X, Wang Y, Zidichouski J ZJ. silibinin: a novel inhibitor of AB aggregation. Neurochemistry international journal. 2011;58:399–403.
  7. Yarnell E, Abascal K. Silybum marianum and silymarin. Vol. 6, Integrative Medicine. 2007. p. 21–5.
  8. FH, A.R. H, S.M. A. A review of herbal medicine: Silybum marianum. Vol. 3, Journal of Medicinal Plants. 2004. p. 14.
  9. Martinelli T, Whittaker A, Benedettelli S, Carboni A, Andrzejewska J. The study of flavonolignan association patterns in fruits of diverging Silybum marianum (L.) Gaertn. chemotypes provides new insights into the silymarin biosynthetic pathway. Phytochemistry. 2017;
  10. Polyak SJ, Morishima C, Lohmann V, Pal S, Lee DYW, Liu Y, et al. Identification of hepatoprotective flavonolignans from silymarin. Proc Natl Acad Sci U S A. 2010;107(13):5995–9.
  11. Tuckar D, Vrsalovic M, Kustrak Danica. Thin-layer chromatographic analysis of milk thistle preparations - {Silybum} marianum ({L}.) {Gaertn}. Farm Glas. 2003;59(Copyright (C) 2016 American Chemical Society (ACS). All Rights Reserved.):303–10.
  12. Hasanloo T, Khavari-Negad R, Majidi E, Ziai SA, Shams-Ardekani MR. Determination of flavonolignans of dried fruits of Silybum marianum (L.) Gaertn. collected from different areas of Iran by spectrophotometry, TLC and HPLC. Journal of Medicinal Plants. 2005;4(SUPPL. 1):25–32.
  13. ICH Topic Q2 (R1) Validation of Analytical Procedures : Text and Methodology. International Conference on Harmonization. 2005;1994(November 1996):17.
  14. Ich Harmonised Tripartite Guideline, Pharmaceutical Development Q8(R2). ICH Harmonised Tripartite Guideline. 2009;8(August):1–28.
  15. Tuckar D, Vrsalovic M, Kustrak Danica. Thin-layer chromatographic analysis of milk thistle preparations - {Silybum} marianum ({L}.) {Gaertn}. Farm Glas. 2003;59(Copyright (C) 2016 American Chemical Society (ACS). All Rights Reserved.):303–10.
  16. Hasanloo T, Khavari-Negad R, Majidi E, Ziai SA, Shams-Ardekani MR. Determination of flavonolignans of dried fruits of Silybum marianum (L.) Gaertn. collected from different areas of Iran by spectrophotometry, TLC and HPLC. Journal of Medicinal Plants. 2005;4(SUPPL. 1):25–32.
  17. Csupor D, Csorba A, Hohmann J. Recent advances in the analysis of flavonolignans of Silybum marianum. J Pharm Biomed Anal. 2016;130:301–17.
  18. Kuki Á, Biri B, Nagy L, Deák G, Kalmár J, Mándi A, et al. Collision induced dissociation study of the major components of silymarin. Int J Mass Spectrom. 2012;315:46–54.
  19. Fenclova M, Stranska-Zachariasova M, Benes F, Novakova A, Jonatova P, Kren V, et al. Liquid chromatography–drift tube ion mobility–mass spectrometry as a new challenging tool for the separation and characterization of silymarin flavonolignans. Anal Bioanal Chem. 2020;412(4):819–32.
  20. Wallace S, Carrier DJ, Beitle RR, Clausen EC, Griffis CL. HPLC-UV and LC-MS-MS Characterization of Silymarin in Milk Thistle Seeds and Corresponding Products. Journal of Nutraceuticals, Functional and Medical Foods. 2003;4(2):37–48.
  21. Zhang C xiao, Chen H, Liu Y bin, Yang X juan, Chen Jun. Comparative study on the extraction of silymarin and silybin from {Silybum} marianum dregs and {Silybum} marianum shells. Zhongchengyao. 2013;35(Copyright (C) 2016 American Chemical Society (ACS). All Rights Reserved.):2137–41.
  22. Saleh IA, Vinatoru M, Mason TJ, Abdel-Azim NS, Aboutabl EA, Hammouda FM. Ultrasonic-assisted extraction and conventional extraction of silymarin from Silybum marianum seeds; A comparison. Res J Pharm Biol Chem Sci. 2015;6(2):709–17.
  23. AbouZid SF, Chen SN, Pauli GF. Silymarin content in Silybum marianum populations growing in Egypt. Ind Crops Prod. 2016.
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