Journal of Natural Medicines

, Volume 67, Issue 3, pp 554–561 | Cite as

Comparative study of three Plumbago L. species (Plumbaginaceae) by microscopy, UPLC-UV and HPTLC

  • Ahmed M. Galal
  • Vijayasankar Raman
  • Bharathi Avula
  • Yan-Hong Wang
  • Chidananda Swamy Rumalla
  • Aruna Dharmapriya Weerasooriya
  • Ikhlas A. Khan
Original Paper


This paper presents a comparative study of anatomy of leaves, stems and roots of three species of Plumbago, namely P. auriculata Lam., P. indica L. and P. zeylanica L. by light microscopy. The paper also provides qualitative and quantitative analysis of the naphthoquinone, plumbagin—a major constituent present in these species—using UPLC-UV. Microscopic examinations revealed the presence of distinctive differences in the anatomical features of the leaf, stem and root of the three species, and these can thus be used for identification and authentication of these species. UPLC-UV analysis showed the highest concentration of plumbagin in the roots of P. zeylanica (1.62 % w/w) followed by the roots of P. indica (0.97 % w/w) and then P. auriculata (0.33–0.53 % w/w). In contrast, plumbagin was not detected in the stems and leaves of P. indica and in the leaves of P. auriculata, whereas very low concentrations (<0.02 % w/w) of plumbagin were detected in the stems and leaves of P. zeylanica and in the stems of P. auriculata. HPTLC fingerprints of the leaf and root of the three species exhibited distinguishable profiles, while those of the stems were undifferentiated.


Plumbago Plumbaginaceae Anatomy Microscopy Plumbagin UPLC-UV HPTLC 


  1. 1.
    Mabberley DJ (1997) The plant-book. A portable dictionary of the vascular plants. Cambridge University Press, CambridgeGoogle Scholar
  2. 2.
    Kini DP, Pandey S, Shenoy BD, Singh UV, Udupa N, Umadevi P, Kamath R, Nagarajkumari, Ramanarayan K (1997) Antitumor and antifertility activities of plumbagin controlled release formulations. Indian J Exp Biol 35:374–379PubMedGoogle Scholar
  3. 3.
    Gopinath S, Muralidharan S, Rajan S, Danaphal SP (2009) Simultaneous estimation of plumbagin and embelin by reverse phase-high performance liquid chromatographic method. Pharm Lett 1:135–142Google Scholar
  4. 4.
    Krishnaswamy M, Purushothaman KK (1980) Plumbagin: a study of its anticancer, antibacterial and antifungal properties. Indian J Exp Biol 18:876–877PubMedGoogle Scholar
  5. 5.
    Tilak JC, Adhikari S, Devasagayam TPA (2004) Antioxidant properties of Plumbago zeylanica, an Indian medicinal plant and its active ingredient, plumbagin. Redox Rep 9:219–227PubMedCrossRefGoogle Scholar
  6. 6.
    Nile SH, Khobragade CN (2010) Antioxidant activity and flavonoid derivatives of Plumbago zeylanica. J Nat Prod 3:130–133Google Scholar
  7. 7.
    Van dVLM (1972) Distribution of plumbagin in the Plumbaginaceae. Phytochemistry 11:3247–3248CrossRefGoogle Scholar
  8. 8.
    Gullett NP, Ruhul AARM, Bayraktar S, Pezzuto JM, Shin DM, Khuri FR, Aggarwal BB, Surh Y-J, Kucuk O (2010) Cancer prevention with natural compounds. Semin Oncol 37:258–281PubMedCrossRefGoogle Scholar
  9. 9.
    Chatterjee A, Pakrashi SC (1997) The treatise on Indian medicinal plants. Council of Scientific and Industrial Research, New DelhiGoogle Scholar
  10. 10.
    Sharma PC, Yelne MB, Dennis TJ (2000) Database on medicinal plants used in ayurveda (on CD). Central Council for Research in Ayurveda and Siddha, New DelhiGoogle Scholar
  11. 11.
    Ajayi GO, Olagunju JA, Ademuyiwa O, Martins OC (2011) Gas chromatography-mass spectrometry analysis and phytochemical screening of ethanolic root extract of Plumbago zeylanica Linn. J Med Plant Res 5:1756–1761Google Scholar
  12. 12.
    Dorni AIC, Vidyalakshmi KS, Vasanthi HR, Rajamanickam GV (2010) HPTLC method for the quantification of plumbagin in three Plumbago species. Res J Phytochem 4:207–212Google Scholar
  13. 13.
    Pawar RK, Sharma S, Singh KC, Sharma RK (2010) HPTLC method for the determination of Plumbagin from Plumbago zeylanica Linn. (root). Int J Pharm Pharm Sci 2:219–223Google Scholar
  14. 14.
    Mallavadhani UV, Sahu G, Muralidhar J (2002) Screening of Plumbago species for the bioactive marker plumbagin. Pharm Biol 40:508–511CrossRefGoogle Scholar
  15. 15.
    Pawar RK, Sharma S, Singh KC, Sharma RKR (2011) Physico-chemical standardisation and development of HPTLC method for the determination of Plumbagin in Kalmegh Navayas Loha-an ayurvedic formulation. Int J Curr Pharm Res 3:43–48Google Scholar
  16. 16.
    Sasikumar M, Sriram SK (2010) HPTLC analysis of various market samples of a traditional drug source—Kodiveli (Plumbago zeylanica Linn.). Int J Pharm Pharm Sci 2:130–132Google Scholar
  17. 17.
    Dubey N, Dubey N, Mehta R, Saluja AK (2009) Determination of psoralen and plumbagin from its polyherbal oil formulations by an HPTLC densitometric method. J AOAC Int 92:779–784PubMedGoogle Scholar
  18. 18.
    Michelitsch A, Wurglics M, Schubert-Zsilavecz M, Likussar W (1999) Determination of 5-hydroxynaphthoquinones in phytotherapeutic Drosera preparations by differential pulse polarography. Phytochem Anal 10:64–68CrossRefGoogle Scholar
  19. 19.
    Meyyanathan SN, Rajan S, Muralidharan S, Satyanarayana IVV, Suresh B (2009) Quantification of plumbagin in Plumbago zeylanica L. by RP-HPLC method. Int J Recent Prog Medicinal Plants 24:61–67Google Scholar
  20. 20.
    Wang Y-C, Huang T-L (2005) High-performance liquid chromatography for quantification of plumbagin, an anti-Helicobacter pylori compound of Plumbago zeylanica L. J Chromatogr A 1094:99–104PubMedCrossRefGoogle Scholar
  21. 21.
    Gupta MM, Verma RK, Uniyal GC, Jain SP (1993) Determination of plumbagin by normal-phase high-performance liquid chromatography. J Chromatogr 637:209–212CrossRefGoogle Scholar
  22. 22.
    Marston A, Hostettmann K (1984) High-performance liquid chromatography of some naturally occurring naphthoquinones. J Chromatogr 295:526–529CrossRefGoogle Scholar
  23. 23.
    Stensen W, Jensen E (1994) High-performance liquid chromatographic separations of naphthoquinones and their derivatives. Effect of hydrogen bonding on retention. J Chromatogr A 659:87–93CrossRefGoogle Scholar
  24. 24.
    Hsieh Y-J, Lin L-C, Tsai T-H (2005) Determination and identification of plumbagin from the roots of Plumbago zeylanica L. by liquid chromatography with tandem mass spectrometry. J Chromatogr A 1083:141–145PubMedCrossRefGoogle Scholar
  25. 25.
    Babula P, Mikelova R, Adam V, Kizek R, Havel L, Sladky Z (2006) Using of liquid chromatography coupled with diode array detector for determination of naphthoquinones in plants and for investigation of influence of pH of cultivation medium on content of plumbagin in Dionaea muscipula. J Chromatogr B Anal Technol Biomed Life Sci 842:28–35CrossRefGoogle Scholar
  26. 26.
    Hsieh Y-J, Lin L-C, Tsai T-H (2006) Measurement and pharmacokinetic study of plumbagin in a conscious freely moving rat using liquid chromatography/tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 844:1–5CrossRefGoogle Scholar
  27. 27.
    Chamberlain CJ (1901) Methods in plant histology. University of Chicago Press, ChicagoGoogle Scholar
  28. 28.
    Ruzin SE (1999) Plant microtechnique and microscopy. Oxford University Press, New YorkGoogle Scholar
  29. 29.
    ICH (2005) Validation of analytical procedures: text and methodology. ICH harmonized tripartite guidelinesGoogle Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan 2012

Authors and Affiliations

  • Ahmed M. Galal
    • 1
  • Vijayasankar Raman
    • 1
  • Bharathi Avula
    • 1
  • Yan-Hong Wang
    • 1
  • Chidananda Swamy Rumalla
    • 1
  • Aruna Dharmapriya Weerasooriya
    • 1
  • Ikhlas A. Khan
    • 1
    • 2
  1. 1.National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of PharmacyUniversity of MississippiUniversityUSA
  2. 2.Department of Pharmacognosy, Research Institute of Pharmaceutical Sciences, School of PharmacyUniversity of MississippiUniversityUSA

Personalised recommendations