Skip to main content

Advertisement

SpringerLink
Log in

Metabolite Profiling of In Vitro Cultured and Field Grown Rhizomes of Acorus calamus from Mongolia Using GC–MS

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

Acorus calamus (sweet flag) is used in the traditional Chinese and Indian medicines for various ailments. Due to its extensive use in herbal medicine, natural resources from the world’s forests are being depleted at an alarming rate. In the present study, an in vitro cell culture technique is being explored as an alternative to field grown A. calamus with respect to the metabolite profile, antioxidant properties, total phenol, and total flavonoid content. Gas chromatography mass spectrometry (GC–MS) was utilized to compare the metabolite profiling between methanolic extracts of in vitro and field grown rhizome tissues of A. calamus. A statistical analysis indicated an upregulation of α-selinene, which is representative of sesquiterpene ketones, and a cyclic polyol, d-pinitol, which has an insulin mimicking effect in the in vitro cultivated rhizome tissue when compared to field grown rhizomes. Significantly higher free-radical scavenging activity (IC50 69.32 μg mL−1), total phenolic content (71.60 mg GAE g−1), and total flavonoid content (42.34 mg CE g−1) were observed in in vitro rhizome tissues compared with those from field grown rhizomes. These observations suggest that the in vitro cultivation of Acorus rhizomes could be exploited as an alternative to field grown A. calamus, as it is an endangered medicinal plant. The production of useful metabolites by the in vitro cultured rhizomes can be explored successfully for utilization by various food and drug industries.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Kirtikar KR, Basu BD (1987) Indian Medicinal Plants. Mahendra Pal Sing Publishers, Dehradun

    Google Scholar 

  2. Sharma Vikas, Singh I, Chaudhary P (2014) Acorus calamus (the healing plant): a review on its medicinal potential, micropropagation and conservation. Nat Prod Res 28:1454–1466

    Article  CAS  Google Scholar 

  3. Imam H, Riaz Z, Azhar M, Sofi G, Hussain A (2013) Sweet flag (Acorus calamus Linn.): An incredible medicinal herb. Int J Green Pharm 7:288

  4. Raina V, Srivastava S, Syamasunder K (2003) Essential oil composition of Acorus calamus L. from the lower region of the Himalayas. Flavour Fragr J 18:18–20

    Article  CAS  Google Scholar 

  5. Verma S, Singh N (2012) In vitro mass multiplication of Acorus calamus L.: an endangered medicinal plant. Am-Euras J Agric Environ Sci 12:1514–1521

    CAS  Google Scholar 

  6. Hettiarachchi A, Fernando KKS, Jayasuriya AHM (2012) In vitro propagation of wadakaha (Acorus calamus L.). J Natl Sci Found 25:151–157

    Google Scholar 

  7. Hall R, Beale M, Fiehn O, Hardy N, Sumner L, Bino R (2002) Plant metabolomics the missing link in functional genomics strategies. Plant Cell 14:1437–1440

    Article  CAS  Google Scholar 

  8. Lim CK, Lord GA (2002) Current developments in LCMS for pharmaceutical analysis. Biol Pharm Bull 25:547–557

    Article  CAS  Google Scholar 

  9. Pasikanti KK, Ho PC, Chan ECY (2008) GC–MS in metabolic profiling of biological fluids. J Chromatogr B 871:202–211

    Article  CAS  Google Scholar 

  10. Lenz EM, Wilson ID (2007) Analytical strategies in metabonomics. J Proteome Res 6:443–458

    Article  CAS  Google Scholar 

  11. Aharoni A, Ric de Vos CH, Verhoeven HA, Maliepaard C, Kruppa G, Bino R, Goodenowe DB (2002) Nontargeted metabolome analysis byu use of FTICRMS. OMICS J Integr Biol 6:217–234

    Article  CAS  Google Scholar 

  12. Powers R (2009) NMR metabolomics and drug discovery. Magn Reson Chem 47:S2–S11

    Article  CAS  Google Scholar 

  13. Lu H, Dunn WB, Shen H, Kell DB, Liang Y (2008) Comparative evaluation of software for deconvolution of metabolomics data based on GC ToF MS. Trends Anal Chem 27:215–227

    Article  CAS  Google Scholar 

  14. Dai J, Mumper RJ (2010) Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules 15:7313–7352

    Article  CAS  Google Scholar 

  15. Senthil K, Thirugnanasambantham P, Oh TJ, Kim SH, Choi HK (2015) Free Radical Scavenging Activity and Comparative Metabolic Profiling of In vitro Cultured and Field Grown Withania somnifera Roots. PLoS One e0123360

  16. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  17. Vaz JA, Barros L, Martins A, Santos-Buelga C, Vasconcelos MH, Ferreira ICFR (2011) Chemical composition of wild edible mushrooms and antioxidant properties of their water soluble polysaccharidic and ethanolic fractions. Food Chem 126:610–616

    Article  CAS  Google Scholar 

  18. Hithamani G, Srinivasan K (2014) Effect of domestic processing on the polyphenol content and bioaccessibility in finger millet (Eleusine coracana) and pearl millet (Pennisetum glaucum). Food Chem 164:55–62

    Article  CAS  Google Scholar 

  19. Lahouar L, Arem AE, Ghrairi F, Chahdoura H, Ben SH, Felah ME (2014) Phytochemical content and antioxidant properties of diverse varieties of whole barley (Hordeum vulgare L.) grown in Tunisia. Food Chem 145:578–583

    Article  CAS  Google Scholar 

  20. Rolland F, Moore B, Sheen J (2002) Sugar sensing and signaling in plants. Plant Cell 14:S185–S205

    CAS  Google Scholar 

  21. Sengun IY, Karabiyikli S (2011) Importance of acetic acid bacteria in food industry. Food Control 22:647–656

    Article  CAS  Google Scholar 

  22. Omura Y, O’Young B, Jones M, Pallos A, Duvvi H, Shimotsuura Y (2011) Caprylic acid in the effective treatment of intractable medical problems of frequent urination, incontinence, chronic upper respiratory infection, root canalled tooth infection, ALS, etc., caused by asbestos & mixed infections of Candida albicans, Helicobacter pylori & Cytomegalovirus with or without other microorganisms & mercury. Acupunct Electrother Res 36:19–64

    Article  Google Scholar 

  23. Ashaari Z, Hanim R, Tahir PM, Nizam N (2004) Effects of peroxide and oxalic acid bleaching on the colour and gluing properties of some tropical bamboos. J Biol Sci 4:90–94

    Article  Google Scholar 

  24. Spina A, Sorvillo L, Esposito A, Borgia A, Sapio L, Naviglio S (2012) Inorganic phosphate as a signaling molecule: a potential strategy in osteosarcoma treatment. Curr Pharm Des 19:5394–5403

    Article  Google Scholar 

  25. Wald SL, McLaurin RL (1982) Oral glycerol for the treatment of traumatic intracranial hypertension. J Neurosurg 56:323–331

    Article  CAS  Google Scholar 

  26. Cok B, Tsiropoulos I, Roes AL, Patel MK (2014) Succinic acid production derived from carbohydrates: an energy and greenhouse gas assessment of a platform chemical toward a bio-based economy. Biofuels Bioprod Biorefin 8:16–29

    Article  CAS  Google Scholar 

  27. Bizzari, Sebastian N, Blagoev, Milen, “Propionic Acid,” CEH Marketing Research Report: Chemical Economics Handbook, SRI Consulting, November 2007, pages 6, and 14–16

  28. Kasumyan AO, Døving KB (2003) Taste preferences in fishes. Fish Fish 4:289–347

    Article  Google Scholar 

  29. Devi SA, Mali AL, Rahee MA, Belinda EDS (2014) Antioxidant properties of Alpha asarone. Asian J Biochem 9:107–113

    Article  CAS  Google Scholar 

  30. Kircheis G, Nilius R, Held C, Berndt H, Buchner M, Gortelmeyer R, Hendricks R, Kruger B, Kuklinski B, Meister H, Otto HJ (1997) Therapeutic efficacy of l-ornithine-l-aspartate infusions in patients with cirrhosis and hepatic encephalopathy: results of a placebo-controlled, double-blind study. Hepatology 25:1351–1360

    Article  CAS  Google Scholar 

  31. Bader A, Caponi C, Cioni PL, Flamini G, Morelli I (2003) Acorenone in the essential oil of flowering aerial parts of Seseli tortuosum L. Flavour and fragrance journal 18:57–58

    Article  CAS  Google Scholar 

  32. Olguín HJ, Guzmán DC, García EH, Mejía GB (2016) The Role of Dopamine and Its Dysfunction as a Consequence of Oxidative Stress. Oxid Med Cell Longev 2016(11):1–13

    Article  Google Scholar 

  33. Govindarajan C, Pitchaipillai R, Shanmugasundaram B, Thangam S, Arokiasamy J, Pillai MS (2015) Myoinositol: A review of its use in patients with polycystic ovary syndrome. World J Pharm Pharmaceut Sci 4:137–155

    CAS  Google Scholar 

  34. Bernstein AM, Roizen MF, Martinez L (2014) Purified palmitoleic acid for the reduction of high-sensitivity C-reactive protein and serum lipids: a double-blinded, randomized, placebo controlled study. J Clin Lipidol 8:612–617

    Article  Google Scholar 

  35. Kris-Etherton PM, Griel AE, Psota TL, Gebauer SK, Zhang J, Etherton TD (2005) Dietary stearic acid and risk of cardiovascular disease: intake, sources, digestion, and absorption. Lipids 40:1193–1200

    Article  CAS  Google Scholar 

  36. Bates SH, Jones RB, Bailey CJ (2000) Insulin-like effect of pinitol. Br J Pharmacol 130:1944–1948

    Article  CAS  Google Scholar 

  37. Vernon D, Bohnert HJ (1992) A novel methyl transferase induced by osmotic stress in the facultative halophyte Mesembryanthemum crystallinum. EMBO J 11:2077–2086

    CAS  Google Scholar 

  38. Lau VW, Journoud M, Jones PJ (2005) Plant sterols are efficacious in lowering plasma LDL and non-HDL cholesterol in hypercholesterolemic type 2 diabetic and nondiabetic persons. Am J Clin Nutri 81:1351–1358

    CAS  Google Scholar 

  39. Raskin I, Ribnicky DM, Komarnytsky S, Ilic N, Poulev A, Borisjuk N, Brinker A, Moreno DA, Ripoll C, Yakoby N, O’Neal JM (2002) Plants and human health in the twenty-first century. Trends Biotechnol 20:522–531

    Article  CAS  Google Scholar 

  40. Ponchet J, Martin-Tanguy A, Marais Martin MC (1982) Hydroxycinnamoyl acid amides and aromatic amines in the inflorescences of some Araceaespecies. Phytochem 21:2865–2869

    Article  CAS  Google Scholar 

  41. Kulma A, Szopa J (2007) Catecholamines are active compounds in plants. Plant Sci 172:433–440

    Article  CAS  Google Scholar 

  42. WuH Fang Y (2004) Pharmacokinetics of beta asarone in rats. Acta Pharm Sin 39:836–838

    Google Scholar 

  43. Lander V, Schreier P (1990) Acorenone and c asarone: indicators of the origin of calamus oils (A. calamus L.). Flav Fragr J 5:75–79

    Article  CAS  Google Scholar 

  44. Venskutonis P, Dagilyte A (2003) Composition of essential oil of sweet flag leaves at different growing phases. J Essent Oil Res 15:313–318

    Article  CAS  Google Scholar 

  45. Gretšušnikova T, Koel M, Orav A (year not mentioned) Comparison of the essential oil composition of Acorus calamus obtained by supercritical carbon dioxide extraction and hydrodistillation methods

  46. Jaiswal Y, Liang Z, Ho A, Chen H, Zhao Z (2015) Metabolite Profiling of Tissues of Acorus calamus and Acorus tatarinowii rhizomes by using LMD, UHPLC-QTOF MS, and GC-MS. Planta Med 81:333–341

    Article  CAS  Google Scholar 

  47. Mazza G (1985) Gas chromatographic and mass spectrometric studies of the constituents of the rhizome of calamus. I. the volatile constituents of the essential oil. J Chromatogr A 328:179–194

    Article  CAS  Google Scholar 

  48. Shailajan S, Menon S, Swar G, Singh D, Nair S (2015) Estimation and quantitation of β-asarone from Acorus calamus rhizome and its formulations using validated RP-HPLC method. Pharm Methods 6:94

    Article  Google Scholar 

  49. Wua H, Sia M, Loua J, Zhoub C, Shena J, Wua H, Yanga B, Hea Q (2010) Insulin releasing and alpha-glucosidase inhibitory activity of ethyl acetate fraction of Acorus calamus in vitro and in vivo. J Ethnopharmacol 128:154–159

    Article  Google Scholar 

  50. Mijares AH, Banuls C, Peris JE, Monzo N, Jover A, Bellod L, Victor VM, Rocha M (2013) A single acute dose of pinitol from a naturally-occurring food ingredient decreases hyperglycaemia and circulating insulin levels in healthy subjects. Food Chem 141:1267–1272

    Article  Google Scholar 

  51. Sivakumar S, Subramanian SP (2009) Pancreatic tissue protective nature of d-pinitol studied in streptozotocin-mediated oxidative stress in experimental diabetic rats. Eur J Pharmacol 622:65–70

    Article  CAS  Google Scholar 

  52. Kim MJ, Yoo KH, Kim JH, Seo YT, Ha BW, Kho JH, Shin YG, Chung CH (2007) Effect of pinitol on glucose metabolism and adipocytokines in uncontrolled type 2 diabetes. Diabetes Res Clin Pract 77S:S247–S251

    Article  Google Scholar 

  53. Szoke E, Maday E, Marczal G, Lemberkovics E (2003) Analysis of biologically active essential oil components of chamomiles in Hungary (In vivo–In vitro). Acta horticulturae

  54. MacLeod G, Ames JM (1989) Volatile components of celery and celeriac. Phytochem 28:1817–1824

    Article  CAS  Google Scholar 

  55. Li XG, Zhang TL, Wang XX, Hua K, Zhao L, Ham ZM (2013) The composition of root exudates from two different resistant peanut cultivars and their effects on the growth of soil-borne pathogen. Int J Biol Sci 9:164–173

    Article  Google Scholar 

  56. Denton MD, Veneklaas EJ, Lambers H (2007) Does phenotypic plasticity in carboxylate exudation differ among rare and widespread Banksia species (Proteacea)? New Phytol 173:592–599

    Article  CAS  Google Scholar 

  57. Kedare SB, Singh RP (2011) Genesis and development of DPPH method of antioxidant assay. J Food Sci Technol 48:412–422

    Article  CAS  Google Scholar 

  58. Devi SA, Ganjewala D (2011) Antioxidant activities of methanolic extracts of sweet flag (Acorus calamus) leaves and rhizomes. J Herbs Spiced Med Plants 17:1–11

    Article  CAS  Google Scholar 

  59. Elayaraja A, Vijayalakshmi M, Devalarao G (2010) In vitro free radical scavenging activity of various root and rhizome extracts of Acorus calamus Linn. Int J Pharm Biol Sci 1:301–304

    Google Scholar 

  60. Nikolova M (2011) Screening of radical scavenging activity and polyphenol content of Bulgarian plant species. Pharmacogn Res 3:256

    Article  CAS  Google Scholar 

  61. Pereira DM, Valentão P, Pereira JA, Andrade PB (2009) Phenolics: From chemistry to biology. Molecules 14:2202–2211

    Article  CAS  Google Scholar 

Download references

Acknowledgments

PDD acknowledges the Department of Biotechnology-IISc Grant-in-aid Partnership Programme for financial support. We thank Steve Madden for providing the Mass Profiler Professional MPP software. The MPP software was supported by Durairaj, Strand Life Sciences Pvt Ltd., and MPP software team of Agilent Technologies is greatly appreciated. The financial grant under the Research Training Fellowship for Developing Country Scientists (RTF-DCS), Centre for Science and Technology of the Non-Aligned, and Other Developing Countries to OK is gratefully acknowledged.

Author information

Author notes

    Authors and Affiliations

    1. GC–MS Facility, Division of Biological Sciences, Indian Institute of Science, Bengaluru, 560 012, India

      Putchen Dakshinamoorthy Deepalakshmi

    2. Laboratory of Plant Biotechnology, Institute of General and Experimental Biology, Mongolian Academy of Sciences, Ulaanbaatar, 13330, Mongolia

      Khongorzul Odgerel, Oyunbileg Yungeree & Altanzul Khorolragchaa

    3. Department of Biochemistry, Biotechnology and Bioinformatics, Avinashilingam University for Women, Coimbatore, 641 043, India

      Khongorzul Odgerel, Pankajavalli Thirugnanasambantham & Kalaiselvi Senthil

    Authors
    1. Putchen Dakshinamoorthy Deepalakshmi
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Khongorzul Odgerel
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Pankajavalli Thirugnanasambantham
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Oyunbileg Yungeree
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Altanzul Khorolragchaa
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Kalaiselvi Senthil
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Kalaiselvi Senthil.

    Ethics declarations

    Funding

    This work is funded by the Department of Biotechnology—IISc Grant-in-aid Partnership Programme and Research Training Fellowship for Developing Country Scientists (RTF-DCS), Centre for Science & Technology of the Non-Aligned. and Other Developing Countries.

    Conflict of interest

    We declare that we have no conflicts of interest in the authorship or publication of this contribution.

    Ethical approval

    This article does not contain any studies with human participants or animals performed by any of the authors.

    Additional information

    P. D. Deepalakshmi and P. Thirugnanasambantham contributed equally to this work.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    Supplementary material 1 (PDF 1687 kb)

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Deepalakshmi, P.D., Odgerel, K., Thirugnanasambantham, P. et al. Metabolite Profiling of In Vitro Cultured and Field Grown Rhizomes of Acorus calamus from Mongolia Using GC–MS. Chromatographia 79, 1359–1371 (2016). https://doi.org/10.1007/s10337-016-3152-7

    Download citation

    • Received:

    • Revised:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s10337-016-3152-7

    Keywords

    Search

    Navigation