Rapid validated high performance thin layer chromatography method for simultaneous estimation of mangiferin and scopoletin in Canscora decussata (South Indian Shankhpushpi) extract

Abstract

Mangiferin (polyphenolic xanthone) and scopoletin (phenolic coumarin) are well-studied biological markers present in Canscora decussata (Roxb.) Roem. & Schult., Gentianaceae. The objective set for the present studies is to establish and develop a new, simple, selective, sensitive, and precise high performance thin layer chromatography method for the simultaneous estimation of mangiferin and scopoletin in hydroalcoholic extract of C. decussata. The thin layer chromatographic separation of these biomarkers was carried out on aluminum plate pre-coated with silica gel 60F254, eluted with ethyl acetate:acetic:acid:formic acid:water (10:0.5:0.5:1.5). The plate was then dried and densitometric scanning was performed at 254 nm using a Camag TLC scanner III. The system was found to give compact spots for mangiferin (RF0.22) and scopoletin (RF0.78). A good relationship of linear precision between the concentrations (100–600 ng/spot) and peak areas was obtained with correlation coefficient (r) of 0.9979 (mangiferin) and 0.9962 (scopoletin), respectively. The limits of detection and limit of quantification were determined to be 46 and 94 ng/spot for mangiferin and 31 and 78 ng/spot for scopoletin respectively. The percentage of recovery was found from 99.91 to 99.94% for mangiferin and 99.75 to 99.86% for scopoletin. Results obtained from recovery studies showed excellent reliability and reproducibility of the method. Present communication on validated high performance thin layer chromatography method may provide a new, selective, sensitive, and precise method to estimate mangiferin and scopoletin as phytomarkers in the hydroalcoholic extract of C. decussata used in Ayurvedic formulations.© 2015 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. All rights reserved.

References

  1. Abourashed, E.A., Mossa, J.S., 2004. HPTLC determination of caffeine in stimulant herbal products and power drinks. J. Pharm. Biomed. Anal. 36, 617–620.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Agrawal, H., Kaul, N., Paradkar, A.R., Mahadik, K.R., 2004. HPTLC method for guggulsterone. I. Quantitative determination of E- and Z-guggulsterone in herbal extract and pharmaceutical dosage form. J. Pharm. Biomed. Anal. 36, 33–34.

    CAS  PubMed  Google Scholar 

  3. Agrawal, R., Sethiya, N.K., Mishra, S.H., 2013. Antidiabetic activity of alkaloids of Aerva lanata roots on streptozotocin-nicotinamide induced type-II diabetes in rats. Pharm. Biol. 51, 635–642.

    CAS  PubMed  Google Scholar 

  4. Bhatt, D., Sethiya, N.K., Mishra, S.H., 2010. Pharmacognostic studies on the aerial parts of Suaeda maritima Linn (Chenopodiaceae). Niger. J. Nat. Prod. Med. 14, 1–5.

    Google Scholar 

  5. Bhatt, M.K., Dholwani, K.K., Saluja, A.K., 2011. Isolation and structure elucidation of scopoletin from Ipomoea reniformis (Convolvulaceae). J. Appl. Pharm. Sci. 1, 138–144.

    Google Scholar 

  6. Bhuvaneswari, K., 2013. Isolation of mangiferin from leaves of Mangifera indica Var Alphonso. Asian J. Pharm. Clin. Res. 6, 173–174.

    CAS  Google Scholar 

  7. Biringanine, G., Chiarelli, M.T., Faes, M., Duez, P., 2006. A validation protocol for the HPTLC standardization of herbal products: application to the determination of acteoside in leaves of Plantago palmata Hook f.s. Talanta 69, 418–424.

    CAS  PubMed  Google Scholar 

  8. Chaudhuri, R.K., Ghosal, S., 1971. Xanthones of Canscora decussata Schult. Phytochemistry 10, 2425–2432.

    CAS  Google Scholar 

  9. Darmawan, A., Kosela, S., Kardono, L.B.S., Syah, Y.M., 2012. Scopoletin, a coumarin derivative compound isolated from Macaranga gigantifolia Merr. J. Appl. Pharm. Sci. 2, 175–177.

    Google Scholar 

  10. Dineshkumar, B., Mitra, A., Manjunatha, M., 2010. Studies on the antidiabetic and hypolipidemic potentials of mangiferin (xanthone glucoside) in streptozotocininduced type 1 and type 2 diabetic model rats. Int. J. Adv. Pharm. Sci. 1, 75–85.

    CAS  Google Scholar 

  11. Goodwin, R.H., Kavanagh, F., 1949. The isolation of scopoletin, a blue-fluorescing compound from oat roots. Bull. Torrey Bot. Club 76, 255–265.

    CAS  Google Scholar 

  12. ICH, 1996. Q2B. Validation of Analytical Procedure: Methodology. International Conference on Harmonization, Geneva.

    Google Scholar 

  13. ICH, 2005. Q2A. Validation of Analytical Procedures: Text and Methodology. International Conference on Harmonization, Geneva.

    Google Scholar 

  14. Jubert, E., Botha, M., Maicu, C., De Beer, D., Manley, M., 2012. Rapid screening methods for estimation of mangiferin and xanthone contents of Cyclopia subternata plant material. S. Afr. J. Bot. 82, 113–122.

    Google Scholar 

  15. Kapadia, N.S., Acharya, N.S., Acharya, S.A., Shah, M.B., 2006. Use of HPTLC to establish a distinct chemical profile for shankhpushpi and for quantification of scopoletin in Convolvulus pluricaulis Choisy and in commercial formulations of Shankhpushpi. J. Planar Chromatogr. Mod. TLC 19, 195–199.

    CAS  Google Scholar 

  16. Kim, C.Y., Ahn, M., Kim, J., 2006. Preparative isolation of mangiferin from Anemarrhena asphodeloides rhizomes by centrifugal partition chromatography. J. Liq. Chromatogr. Relat. Technol. 29, 869–875.

    Google Scholar 

  17. Krivut, B.A., Fedyunina, N.A., Kocherga, S.I., Rusakoya, S.V., 1976. Spectrophotometric determination of mangiferin. Chem. Nat. Compd. 12, 36–38.

    Google Scholar 

  18. Kumar, V., Mukherjee, K., Kumar, S., Mal, M., Mukherjee, P.K., 2008. Validation of HPTLC method for the analysis of taraxerol in Clitorea ternatea. Phytochem. Anal. 19, 244–250.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Luo, F., Lv, Q., Zhao, Y., Hu, G., Huang, G., Zhang, J., Sun, C., Li, X., Chen, K., 2012. Quantification and purification of mangiferin from Chinese Mango (Mangifera indica L.) cultivars and its protective effect on human umbilical vein endothelial cells under H2O2-induced stress. Int. J. Mol. Sci. 13, 11260–11274.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Mukherjee, D., Kumar, N.S., Khatua, T., Mukherjee, P.K., 2010. Rapid validated HPTLC method for estimation of betulinic acid in Nelumbo nucifera (Nymphaeaceae) rhizome extract. Phytochem. Anal. 21, 556–560.

    CAS  PubMed  Google Scholar 

  21. Nahata, A., Dixit, V.K., 2008. Spectrofluorimetric estimation of scopoletin in Evolvulus alsinoides Linn. and Convulvulus pluricaulis Choisy. Indian J. Pharm. Sci. 70, 834–837.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Nahata, A., Patil, U.K., Dixit, V.K., 2010. Effect of Evolvulus alsinoides Linn. on learning behavior and memory enhancement activity in rodents. Phytother. Res. 24, 486–493.

    CAS  PubMed  Google Scholar 

  23. Rastogi, S., Pandey, M.M., Rawat, A.K.S., 2007. A new, convenient method for determination of mangiferin, an anti-diabetic compound, in Mangifera indica L. J. Planar Chromatogr. Mod. TLC 20, 317–320.

    CAS  Google Scholar 

  24. Risner, C.H., 1994. The determination of scopoletin in environmental tobacco smoke by high-performance liquid chromatography. J. Liq. Chromatogr. 17 (12), 2723–2736.

    CAS  Google Scholar 

  25. Sellamuthu, P.S., Arulselvan, P., Muniappan, B.P., Kandasamy, M., 2012. Effect of mangiferin isolated from Salacia chinensis regulates the kidney carbohydrate metabolism in streptozotocin-induced diabetic rats. Asian. Pac. J. Trop. Biomed. 2, S1583–S1587.

    Google Scholar 

  26. Sethiya, N.K., Mishra, S.H., 2014. Investigation of mangiferin, as a promising natural polyphenol xanthone on multiple targets of Alzheimer’s disease. J. Biol. Active Prod. Nat. 4, 111–119.

    CAS  Google Scholar 

  27. Sethiya, N.K., Nahata, A., Dixit, V.K., 2008. Simultaneous spectrofluorimetric determination of scopoletin and mangiferin in a methanol extract of Canscora decussata Schult. Asian J. Tradit. Med. 3, 224–229.

    CAS  Google Scholar 

  28. Sethiya, N.K., Nahata, A., Dixit, V.K., 2009a. Comparative thin layer chromatographic investigations on sources of Shankhpushpi. Pharmacogn. J. 1, 224–226.

    CAS  Google Scholar 

  29. Sethiya, N.K., Nahata, A., Dixit, V.K., 2010a. Anxiolytic activity of Canscora decussata in albino rats. J. Complement. Integr. Med. 7, 19.

    Google Scholar 

  30. Sethiya, N.K., Nahata, A., Dixit, V.K., Mishra, S.H., 2012. Cognition boosting effect of Canscora decussata (a South Indian Shankhpushpi). Eur. J. Integr. Med. 4, 113–121.

    Google Scholar 

  31. Sethiya, N.K., Nahata, A., Mishra, S.H., Dixit, V.K., 2009b. An update on Shankhpushpi a cognition boosting Ayurvedic medicine. Zhong Xi Yi Jie He Xue Bao 7, 1001–1022.

    CAS  PubMed  Google Scholar 

  32. Sethiya, N.K., Patel, M.B., Mishra, S.H., 2010b. Phytopharmacologic aspects of Canscora decussata Roem and Schult. Pharmacogn. Rev. 4, 49–57.

    PubMed  PubMed Central  Google Scholar 

  33. Sethiya, N.K., Raja, M.K.M.M., Mishra, S.H., 2013. Antioxidant markers based TLCDPPH differentiation on four commercialized botanical sources of Shankhpushpi (medhya rasayana)–a preliminary assessment. J. Adv. Pharm. Technol. Res. 4, 25–30.

    PubMed  PubMed Central  Google Scholar 

  34. Shastry, V., Haldankar, A., Kadam, N., 2009. HPLC estimation of mangiferin in Salacia chinensis Linn. Asian J. Chem. 21, 6679–6682.

    CAS  Google Scholar 

  35. Silva, W.P., Deraniyagala, S.A., Wijesundera, R.L., Karunanayake, E.H., Priyanka, U.M., 2002. Isolation of scopoletin from leaves of Hevea brasiliensis and the effect of scopoletin on pathogens of H. brasiliensis. Mycopathologia 153, 199–202.

    CAS  PubMed  Google Scholar 

  36. Suryawanshi, S., Asthana, R.K., Gupta, R.C., 2007. Simultaneous estimation of mangiferin and four secoiridoid glycosides in rat plasma using liquid chromatography tandem mass spectrometry and its application to pharmacokinetic study of herbal preparation. J. Chromatogr. B: Anal. Technol. Biomed. Life Sci. 858, 211–219.

    CAS  Google Scholar 

  37. Trivedi, A., Sethiya, N.K., Mishra, S.H., 2011. Preliminary pharmacognostic and phytochemical analysis of Granthika (Leonotis nepetaefolia). An Ayurvedic herb. Indian J. Tradit. Know. 10, 682–688.

    Google Scholar 

  38. Tuzimski, T., Bartosiewicz, E., 2003. Correlation of retention parameters of pesticides in normal and RP systems and their utilization for the separation of a mixture of ten urea herbicides and fungicides by two dimensional TLC on cyano propyl-bonded polar stationary phase and two-adsorbent-layer multi-K plate. Chromatographia 58, 781–788.

    CAS  Google Scholar 

  39. Upadhyay, V., Sharma, N., Tiwari, A.K., Joshi, H.M., Malik, A., Singh, B., Kalakoti, B.S., 2013. Standardization of HPLC method of scopoletin in different extracts of Convolvulus pluricaulis. Int. J. Pharm. Sci. Drug Res. 5, 28–31.

    Google Scholar 

  40. Wagner, S., Urena, A., Reich, E., Merfort, I., 2008. Validated HPTLC methods for the determination of salicin in Salix sp. and of harpagoside in Harpogophytum procumbens. J. Pharm. Biomed. Anal. 48, 587–591.

    CAS  PubMed  Google Scholar 

  41. Xia, Y., Dai, Y., Wang, Q., Liang, H., 2007. Determination of scopoletin in rat plasma by high performance liquid chromatographic method with UV detection and its application to a pharmacokinetic study. J. Chromatogr. B: Anal. Technol. Biomed. Life Sci. 857, 332–336.

    CAS  Google Scholar 

Download references

Acknowledgements

The authors thank Laila Impex, Vijayawada, India for the gift sample of standard scopoletin. One of the authors Neeraj K. Sethiya is thankful to University Grants Commission, New Delhi (India) for providing Junior Research Fellowship for the project. Neeraj K. Sethiya and Ashish Trivedi are thankful to Anchrom HPTLC Technologists, Mumbai (India) for providing the facilities for technical advice regarding instrument handling.

Author information

Affiliations

Authors

Contributions

NKS and AT carried out the laboratory work and the data analyses. NKS and SHM wrote the manuscript. SHM designed the study and supervised the laboratory work. All the authors have read the final manuscript and approved the submission.

Corresponding author

Correspondence to Neeraj K. Sethiya.

Ethics declarations

The authors declare no conflicts of interest.

Rights and permissions

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Sethiya, N.K., Trivedi, A. & Mishra, S.H. Rapid validated high performance thin layer chromatography method for simultaneous estimation of mangiferin and scopoletin in Canscora decussata (South Indian Shankhpushpi) extract. Rev. Bras. Farmacogn. 25, 193–198 (2015). https://doi.org/10.1016/j.bjp.2015.04.002

Download citation

Keywords

  • Mangiferin
  • Scopoletin
  • HPTLC
  • Validation