Skip to main content
SpringerLink
Log in

Determination of Bioactive Compounds of Fenugreek (Trigonella foenum-graecum) Seeds Using LC-MS Techniques

  • Protocol
  • First Online:
Legume Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2107))

Abstract

Trigonella foenum-graecum (fenugreek), a legume has been used as spice throughout the world to enhance the sensory quality of foods, is considered to have a high nutraceutical value. The antidiabetic actions of Trigonella seeds have been speculated due to the presence of steroid saponins, alkaloids, and fiber content. Five bioactive compounds, namely, trigonelline, isoorientin, orientin, vitexin, and isovitexin, were detected and quantified by HPLC system. Further rapid detection of bioactive compounds was achieved using high-performance liquid chromatography hybrid electrospray quadrupole time-of-flight mass spectrometric (HPLC-ESI-QTOF-MS/MS) method. Twenty-five compounds were tentatively identified based on their retention time and measured accurate mass. The fragmentation patterns of known compounds were utilized to elucidate their structures. Six of these identified compounds, that is, 4-hydroxyisoleucine, trigonelline, isoorientin, isovitexin, pinitol, and sarsasapogenin, were simultaneously quantified by UHPLC-ESI-MS/MS method under the multiple reactions monitoring (MRM) mode using ultrahigh-performance liquid chromatography hybrid electrospray triple quadrupole linear ion trap mass spectrometer. The analytical method was validated and successfully applied for simultaneous determination of compounds in Fenugreek seeds, which indicated the suitability of the validated method for its quality control.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Snehlata HS, Payal DR (2012) Fenugreek (Trigonella foenum-graecum L.) an overview. Int J Curr Pharm Rev Res 2(4):169–187

    Google Scholar 

  2. Hojjat SS, Hojjat H (2015) Effect of nano silver on seed germination and seedling growth in fenugreek seed. Int J Food Eng 1(2):106–110

    Google Scholar 

  3. Acharya SN, Thomas JE, Basu SK (2008) Fenugreek, an alternative crop for semiarid regions of North America. Crop Sci 48(3):841–853

    Article  CAS  Google Scholar 

  4. Petropoulos GA (2003) Fenugreek: the genus Trigonella. CRC Press, Boca Raton, Florida

    Google Scholar 

  5. Krishnaswamy K (2008) Traditional Indian spices and their health significance. Asia Pac J Clin Nutr 17:265–268

    PubMed  Google Scholar 

  6. Lobo V, Patil A, Phatak A, Chandra N (2010) Free radicals, antioxidants and functional foods: impact on human health Pharmacognosy reviews. Pharmacogn Rev 4(8):118

    Article  CAS  Google Scholar 

  7. Reddy RR, Srinivasan K (2011) Hepatoprotective and antioxidant effect of fenugreek (Trigonella foenum-graecum) seeds in mice under lithogenic condition. J Food Biochem 35(6):1619–1626

    Article  CAS  Google Scholar 

  8. Ribes G, Sauvaire Y, Costa CD, Baccou JC, Loubatieres-Mariani MM (1986) Antidiabetic effects of subfractions from fenugreek seeds in diabetic dogs. Proc Soc Exp Biol Med 182(2):159–166

    Article  CAS  Google Scholar 

  9. Wani SA, Kumar P (2018) Fenugreek: a review on its nutraceutical properties and utilization in various food products. J Saudi Soc Agric Sci 17(2):97–106

    Google Scholar 

  10. Dash BK, Sultana S, Sultana N (2011) Antibacterial activities of methanol and acetone extracts of fenugreek (Trigonella foenum-gracum) and coriander (Coriandrum sativum). Life Sci Med Res 27:1–8

    Google Scholar 

  11. Shabbeer S, Sobolewski M, Anchoori RK, Kachhap S, Hidalgo M, Jimeno A, Davidson NE, Carducci M, Khan SR (2009) Fenugreek: a naturally occurring edible spice as an anticancer agent. Cancer Biol Ther 8(3):272–278

    Article  CAS  Google Scholar 

  12. Yoshinari O, Sato H, Igarashi K (2009) Anti-diabetic effects of pumpkin and its components, trigonelline and nicotinic acid, on Goto-Kakizaki rats. Biosci Biotechnol Biochem 73(5):1033–1041

    Article  CAS  Google Scholar 

  13. Khorshidian N, YousefiAsli M, Arab M, AdeliMirzaie A, Mortazavian AM (2016) Fenugreek: potential applications as a functional food and nutraceutical. Nutr Food Sci Res 3(1):5–16

    Article  CAS  Google Scholar 

  14. Kunnumakkara AB, Koca C, Dey S, Gehlot P, Yodkeeree S, Danda D, Sung B, Aggarwal BB (2009) Traditional uses of spices: an overview. In: Molecular targets and therapeutic uses of spices: modern uses for ancient medicine, vol 2. World Scientific Publishing, Singapore, pp 1–24

    Google Scholar 

  15. Younesy S, Amiraliakbari S, Esmaeili S, Alavimajd H, Nouraei S (2014) Effects of fenugreek seed on the severity and systemic symptoms of dysmenorrhea. J. Reprod. Infertil. 15(1):41

    PubMed  PubMed Central  Google Scholar 

  16. Hakimi S, Mohammad Alizadeh S, Delazar A, Abbasalizadeh F, Bamdad Mogaddam R, Siiahi MR, Mostafa Garabagi P (2006) Probable effects of fenugreek seed on hot flash in menopausal women. J. Med. Plants 3(19):9–14

    Google Scholar 

  17. Bashtian MH, Emami SA, Mousavifar N, Esmaily HA, Mahmoudi M, Poor AHM (2013) Evaluation of fenugreek (Trigonella foenum-graceum L.), effects seeds extract on insulin resistance in women with polycystic ovarian syndrome. Iran J Pharm Res 12(2):475

    Google Scholar 

  18. Suresh P, Kavitha CN, Babu SM, Reddy VP, Latha AK (2012) Effect of ethanol extract of Trigonella foenum graecum (fenugreek) seeds on Freund’s adjuvant-induced arthritis in albino rats. Inflammation 35(4):1314–1321

    Article  CAS  Google Scholar 

  19. Arrak JK, Al-Zyadi AJ (2015) The therapeutic role of alcoholic extract of fenugreek seeds on hypothyroidism state induced by thiourea and some blood parameters in adult male rabbits. Iraqi J Vet Med 39(1):1–7

    Google Scholar 

  20. Avalos-Soriano A, De la Cruz-Cordero R, Rosado JL, Garcia-Gasca T (2016) 4-Hydroxyisoleucine from fenugreek (Trigonella foenum-graecum): effects on insulin resistance associated with obesity. Molecules 21(11):1596

    Article  Google Scholar 

  21. Roberts KT (2011) The potential of fenugreek (Trigonella foenum-graecum) as a functional food and nutraceutical and its effects on glycemia and lipidemia. J Med Food 14(12):1485–1489

    Article  CAS  Google Scholar 

  22. Sharma RD, Sarkar A, Hazra DK, Misra B, Singh JB, Maheshwari BB, Sharma SK (1996) Hypolipidaemic effect of fenugreek seeds: a chronic study in non-insulin dependent diabetic patients. Phytother Res 10(4):332–334

    Article  Google Scholar 

  23. Olaiya CO, Soetan KO (2014) A review of the health benefits of fenugreek (Trigonella foenum-graecum L.): nutritional, biochemical and pharmaceutical perspectives. Am J Soc Issues Humanties 4(2):3–12

    Google Scholar 

  24. Charles DJ (2012) Fenugreek. In: Antioxidant properties of spices, herbs and other sources. Springer, New York, NY, pp 295–303

    Chapter  Google Scholar 

  25. Bahmani M, Shirzad H, Mirhosseini M, Mesripour A, Rafieian-Kopaei M (2016) A review on ethnobotanical and therapeutic uses of fenugreek (Trigonella foenum-graceum L). J Evidence-Based Complementary Altern Med 21(1):53–62

    Article  CAS  Google Scholar 

  26. Belguith-Hadriche O, Bouaziz M, Jamoussi K, El Feki A, Sayadi S, Makni-Ayedi F (2010) Lipid-lowering and antioxidant effects of an ethyl acetate extract of fenugreek seeds in high-cholesterol-fed rats. J Agric Food Chem 58(4):2116–2122

    Article  CAS  Google Scholar 

  27. Nagulapalli Venkata KC, Swaroop A, Bagchi D, Bishayee A (2017) A small plant with big benefits: fenugreek (Trigonella foenum-graecum Linn.) for disease prevention and health promotion. Mol Nutr Food Res 61(6):1600950

    Article  Google Scholar 

  28. Mishra P (2009) Isolation, spectroscopic characterization and molecular modeling studies of mixture of Curcuma longa, ginger and seeds of fenugreek. Int. J. PharmTech Res. 1(1):79–95

    CAS  Google Scholar 

  29. Yadav UC, Baquer NZ (2014) Pharmacological effects of Trigonella foenum-graecum L. in health and disease. Pharm Biol 52(2):243–254

    Article  Google Scholar 

  30. Taylor WG, Zaman MS, Mir Z, Mir PS, Acharya SN, Mears GJ, Elder JL (1997) Analysis of steroidal sapogenins from amber fenugreek (Trigonella foenum-graecum) by capillary gas chromatography and combined gas chromatography/mass spectrometry. J Agric Food Chem 45(3):753–759

    Article  CAS  Google Scholar 

  31. Chopra S, Ahmad FJ, Khar RK, Motwani SK, Mahdi S, Iqbal Z, Talegaonkar S (2006) Validated high-performance thin-layer chromatography method for determination of trigonelline in herbal extract and pharmaceutical dosage form. Anal Chim Acta 577(1):46–51

    Article  CAS  Google Scholar 

  32. Faeste CK, Christians U, Egaas E, Jonscher KR (2010) Characterization of potential allergens in fenugreek (Trigonella foenum-graecum) using patient sera and MS-based proteomic analysis. J Proteome 73(7):1321–1333

    Article  CAS  Google Scholar 

  33. Bebrevska L, Bravo L, Vandervoort J, Pieters L, Vlietinck A, Apers S (2007) Development and validation of an HPLC method for quality control of Pueraria lobata flower. Planta Med 73:1606–1613

    Article  CAS  Google Scholar 

  34. Tistaert C, Dejaegher B, Vander Heyden Y (2011) Chromatographic separation techniques and data handling methods for herbal fingerprints: a review. Anal Chim Acta 690(2):148–161

    Article  CAS  Google Scholar 

  35. Vígh S, Cziaky Z, Sinka LT, Pribac C, Moş L, Turcuş V, Remenyik J, Mathe E (2017) Analysis of phytoconstituent profile of fenugreek–Trigonella Foenuem-graecum L.-seed extracts. Stud Univ Babes-Bolyai, Chem 62:145–166

    Article  Google Scholar 

  36. Benayad Z, Gómez-Cordovés C, Es-Safi N (2014) Characterization of flavonoid glycosides from fenugreek (Trigonella foenum-graecum) crude seeds by HPLC–DAD–ESI/MS analysis. Int J Mol Sci 15(11):20668–20685

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge the Sophisticated Analytical Instrument Facility, CSIR-CDRI, Lucknow, India, where all the mass spectrometric studies were carried out.

Author information

Authors and Affiliations

  1. Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India

    Pratibha Singh, Vikas Bajpai, Vikaskumar Gond & Brijesh Kumar

  2. Academy of Scientific and Innovative Research (AcSIR), New Delhi, India

    Pratibha Singh, Ashok Kumar, Narender Tadigoppula & Brijesh Kumar

  3. Medicinal and Process Chemistry, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India

    Ashok Kumar & Narender Tadigoppula

Authors
  1. Pratibha Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vikas Bajpai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vikaskumar Gond
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ashok Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Narender Tadigoppula
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Brijesh Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Brijesh Kumar .

Editor information

Editors and Affiliations

  1. School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi, India

    Mukesh Jain

  2. Department of Life Sciences, School of Natural Sciences, Shiv Nadar University, Gautam Buddha Nagar, Uttar Pradesh, India

    Rohini Garg

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Singh, P., Bajpai, V., Gond, V., Kumar, A., Tadigoppula, N., Kumar, B. (2020). Determination of Bioactive Compounds of Fenugreek (Trigonella foenum-graecum) Seeds Using LC-MS Techniques. In: Jain, M., Garg, R. (eds) Legume Genomics. Methods in Molecular Biology, vol 2107. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0235-5_21

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0235-5_21

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0234-8

  • Online ISBN: 978-1-0716-0235-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation