Skip to main content
Log in

High-speed homogenization coupled with microwave-assisted extraction followed by liquid chromatography–tandem mass spectrometry for the direct determination of alkaloids and flavonoids in fresh Isatis tinctoria L. hairy root cultures

  • Note
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

A new, simple and efficient analysis method for fresh plant in vitro cultures—namely, high-speed homogenization coupled with microwave-assisted extraction (HSH–MAE) followed by liquid chromatography–tandem mass spectrometry (LC–MS/MS)—was developed for simultaneous determination of six alkaloids and eight flavonoids in Isatis tinctoria hairy root cultures (ITHRCs). Compared with traditional methods, the proposed HSH–MAE offers the advantages of easy manipulation, higher efficiency, energy saving, and reduced waste. Cytohistological studies were conducted to clarify the mechanism of HSH–MAE at cellular/tissue levels. Moreover, the established LC–MS/MS method showed excellent linearity, precision, repeatability, and reproducibility. The HSH–MAE–LC–MS/MS method was also successfully applied for screening high-productivity ITHRCs. Overall, this study opened up a new avenue for the direct determination of secondary metabolic profiles from fresh plant in vitro cultures, which is valuable for improving quality control of plant cell/organ cultures and sheds light on the metabolomic analysis of biological samples.

HSH-MAE-LC-MS/MS opened up a new avenue for the direct determination of alkaloids and flavonoids in fresh Isatis tinctoria hairy root cultures

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

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Hamburger M (2002) Isatis tinctoria – from the rediscovery of an ancient medicinal plant towards a novel anti-inflammatory phytopharmaceutical. Phytochem Rev 1:333–344

    CAS  Google Scholar 

  2. Mohn T, Plitzko I, Hamburger M (2009) A comprehensive metabolite profiling of Isatis tinctoria leaf extracts. Phytochemistry 70:924–934

    Article  CAS  Google Scholar 

  3. Mohn T, Potterat O, Hamburger M (2007) Quantification of active principles and pigments in leaf extracts of Isatis tinctoria by HPLC/UV/MS. Planta Med 73:151–156

    CAS  Google Scholar 

  4. Mohn T, Cutting B, Ernst B, Hamburger M (2007) Extraction and analysis of intact glucosinolates—a validated pressurized liquid extraction/liquid chromatography–mass spectrometry protocol for Isatis tinctoria, and qualitative analysis of other cruciferous plants. J Chromatogr A 1166:142–151

    CAS  Google Scholar 

  5. Mohn T, Suter K, Hamburger M (2008) Seasonal changes and effect of harvest on glucosinolates in Isatis leaves. Planta Med 74:582–587

    CAS  Google Scholar 

  6. Gai QY, Jiao J, Luo M, Wang W, Ma W, Zu YG, Fu YJ (2015) Establishment of high-productive Isatis tinctoria L. hairy root cultures: a promising approach for efficient production of bioactive alkaloids. Biochem Eng J 95:37–47

    CAS  Google Scholar 

  7. Steinmann D, Ganzera M (2011) Recent advances on HPLC/MS in medicinal plant analysis. J Pharmaceut Biomed Anal 55:744–757

    CAS  Google Scholar 

  8. Kole PL, Venkatesh G, Kotecha J, Sheshala R (2011) Recent advances in sample preparation techniques for effective bioanalytical methods. Biomed Chromatogr 25:199–217

    CAS  Google Scholar 

  9. Romanik G, Gilgenast E, Przyjazny A, Kamiński M (2007) Techniques of preparing plant material for chromatographic separation and analysis. J Biochem Biophys Methods 70:253–261

    CAS  Google Scholar 

  10. Chan CH, Yusoff R, Ngoh GC, Kung FWL (2011) Microwave-assisted extractions of active ingredients from plants. J Chromatogr A 1218:6213–6225

    CAS  Google Scholar 

  11. Kursinszki L, Hank H, László I, Szőke É (2005) Simultaneous analysis of hyoscyamine, scopolamine, 6β-hydroxyhyoscyamine and apoatropine in Solanaceous hairy roots by reversed-phase high-performance liquid chromatography. J Chromatogr A 1091:32–39

    CAS  Google Scholar 

  12. Wahby I, Arráez-Román D, Segura-Carretero A, Ligero F, Caba JM, Fernández-Gutiérrez A (2006) Analysis of choline and atropine in hairy root cultures of Cannabis sativa L. by capillary electrophoresis-electrospray mass spectrometry. Electrophoresis 27:2208–2215

    Article  CAS  Google Scholar 

  13. Farhat A, Ginies C, Romdhane M, Chemat F (2009) Eco-friendly and cleaner process for isolation of essential oil using microwave energy: experimental and theoretical study. J Chromatogr A 1216:5077–5085

    CAS  Google Scholar 

  14. Hu ZB, Du M (2006) Hairy root and its application in plant genetic engineering. J Integr Plant Biol 48:121–127

    CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge financial support by the Application Technology Research and Development Program of Harbin (2013AA3BS014), the Fundamental Research Funds for the Central Universities (2572015AA14, 2572014AA06), and the Special Fund of the National Natural Science Foundation of China (31270618).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yu-Jie Fu.

Additional information

Jiao Jiao and Qing-Yan Gai contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 348 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jiao, J., Gai, QY., Zhang, L. et al. High-speed homogenization coupled with microwave-assisted extraction followed by liquid chromatography–tandem mass spectrometry for the direct determination of alkaloids and flavonoids in fresh Isatis tinctoria L. hairy root cultures. Anal Bioanal Chem 407, 4841–4848 (2015). https://doi.org/10.1007/s00216-015-8668-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-015-8668-4

Keywords

Navigation