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RP-HPLC Analysis of Anti-Parkinson’s Drug l-DOPA Content in Mucuna Species from Indian Subcontinent

  • Ravishankar Patil
  • Chetan Aware
  • Swaroopsingh Gaikwad
  • Manisha Rajebhosale
  • Vishwas Bapat
  • Shrirang Yadav
  • Jyoti JadhavEmail author
Research Article
  • 14 Downloads

Abstract

Genus Mucuna has tremendous value in the Indian Ayurveda, and the species have been used for the management of diseases including Parkinson’s disease. Unfortunately, species other than M. pruriens are neglected for scientific investigation. In the present study, quantification of anti-Parkinson’s drug l-DOPA content using HPLC from eighteen different Mucuna samples from Indian subcontinent comprising nine species, five varieties of M. pruriens and a commercial Mucuna powder from Zandu Pharmaceutical Ltd. (MZP) is carried out. Seeds of eighteen Mucuna accessions were analyzed for l-DOPA content using RP-HPLC. The l-DOPA content from different samples is compared with commonly used M. pruriens var utilis. Moreover, possible correlation of protein and carbohydrate with l-DOPA synthesis is hypothesized. The authors found varying l-DOPA content in different species and varieties; and higher l-DOPA containing species were M. macrocarpa (11.6 ± 0.21%), M. sanjappae (10.81 ± 0.2%) and M. atropurpurea (10.80 ± 0.1%). Some of the M. pruriens accessions contained higher level of l-DOPA including M. pruriens var hirsuta (10.2 ± 0.15%), M. pruriens var utilis (Manipur) (9.9 ± 0.15%) and M. pruriens var pruriens (Aurangabad) (9.41 ± 0.07 and 9.5 ± 0.12%). The species, M. gigentea had the lowest l-DOPA content (3.68 ± 0.17%). The strong correlation between l-DOPA and carbohydrate content (R2 = 0.7997) was observed, whereas insignificant correlation (R2 = 0.0277) between protein and l-DOPA content was observed. Higher anti-Parkinson’s drug l-DOPA-containing species were found from India which can be explored further for better extraction of this immense drug. The present study opens a new avenue for metabolite modeling in plant for commercial production of l-DOPA at industrial level.

Keywords

Ayurveda HPLC l-DOPA Mucuna Parkinson’s disease Shikimic acid pathway 

Notes

Acknowledgements

The corresponding author thanks to the Department of Biotechnology, Govt. of India, for funding through DBT-IPLS program (Ref. No.: BT/PR4572/INF/22/147) sanctioned to Department of Biotechnology, Shivaji University, Kolhapur, India. The first author acknowledges SUK-DBT IPLS program and N-PDF funded by SERB (PDF/2016/002075) for providing financial support. The second and third authors acknowledge SUK-DBT IPLS program for the fellowship. Prof. Vishwas Bapat thanks National Academy of Sciences, (NASI) Allahabad, India, for NASI-Honorary Scientist Fellowship.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest to publish this manuscript.

References

  1. 1.
    Jadiya P, Khan A, Sammi S, Kaur S, Mir S, Nazir A (2011) Anti-Parkinsonian effects of Bacopa monnieri: insights from transgenic and pharmacological Caenorhabditis elegans models of Parkinson’s disease. Biochem Biophys Res Commun 413(4):605–610CrossRefGoogle Scholar
  2. 2.
    Thomas B, Flint Beal M (2007) Parkinson’s disease. Hum Mol Genet 16:183–194CrossRefGoogle Scholar
  3. 3.
    Inamdar SA, Surwase SN, Jadhav SB, Bapat VA, Jadhav JP (2013) Statistically optimized biotransformation protocol for continuous production of l-DOPA using Mucuna monosperma callus culture. SpringerPlus 2:570–578CrossRefGoogle Scholar
  4. 4.
    Prakash J, Yadav SK, Chouhan S, Singh SP (2013) Neuroprotective role of Withania somnifera root extract in maneb–paraquat induced mouse model of parkinsonism. Neurochem Res 38(5):972–980CrossRefGoogle Scholar
  5. 5.
    Politisa M, Niccolinia F (2014) Serotonin in Parkinson’s disease. Behav Brain Res 277:136–145CrossRefGoogle Scholar
  6. 6.
    Guggenheim Z (1913) Physiol Chem 88:276CrossRefGoogle Scholar
  7. 7.
    Damodaran M, Ramaswamy R (1937) Isolation of l-3: 4-dihydroxyphenylalanine from the seeds of Mucuna pruriens. Biochem J 31(12):2149CrossRefGoogle Scholar
  8. 8.
    Kofman O (1971) Treatment of Parkinson’s disease with l-DOPA: a current appraisal. Can Med Assoc J 104(6):483–487Google Scholar
  9. 9.
    Vijayakumari K, Siddhuraju P, Janardhanan K (1996) Effects of different post-harvest treatments on anti-nutritional factors in seeds of the tribal pulse, Mucuna pruriens (L.) DC. Int J Food Sci Nutr 47(3):263–272CrossRefGoogle Scholar
  10. 10.
    Aguiyi JC, Igweh AC, Egesie UG, Leoncini R (1999) Studies on possible protection against snake venom using Mucuna pruriens protein immunization. Fitoterapia 70(1):21–24CrossRefGoogle Scholar
  11. 11.
    Amin KMY, Khan MN, Hakim SZR et al (1996) Sexual function improving effect of Mucuna pruriens in sexually normal male rats. Fitoterapia 67(1):53–58Google Scholar
  12. 12.
    Lorenzetti F, Mac Issac S, Arnason JT, Awang DVC, Buckles D (1998) The phytochemistry, toxicology and food potential of velvet bean (cow(h)age, cowitch) (Mucuna adans spp., Fabaceae). In: Buckles D, Eteka A, Osiname O, Galiba M, Galino G (eds) Cover crops in West Africa contributing to sustainable agriculture. IDRC, Ottawa, pp 67–84Google Scholar
  13. 13.
    Brilliant MH (2015) Mining retrospective data for virtual prospective drug repurposing: l-DOPA and age-related macular degeneration. Am J Med 129(3):292–298CrossRefGoogle Scholar
  14. 14.
    Koyanagi T, Katayama T, Suzuki H, Nakazawab H, Yokozeki K, Kumagai H (2005) Effective production of 3,4-dihydroxyphenyl-l-alanine (l-DOPA) with Erwinia herbicola cells carrying a mutant transcriptional regulator TyrR. J Biotechnol 115(3):303–306CrossRefGoogle Scholar
  15. 15.
    Pulikkalpura H, Kurup R, Mathew PJ, Baby S (2015) Levodopa in Mucuna pruriens and its degradation. Sci Rep 5:11078CrossRefGoogle Scholar
  16. 16.
    Rathod B, Patel NM (2014) Development of validated RP-HPLC method for the estimation of l-DOPA from Mucuna pruriens, its extracts and in Aphrodisiac formulation. IJPSR 5:508–513Google Scholar
  17. 17.
    Lowry OH, Rosbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275Google Scholar
  18. 18.
    Trevelyan WE, Forrest RS, Harrison JS (1952) Determination of yeast carbohydrates with the anthrone reagent. Nature 170:626–627CrossRefGoogle Scholar
  19. 19.
    Krishnaveni R, Rathod V, Thakur M, Neelgund Y (2009) Transformation of l-tyrosine to l-DOPA by a novel fungus, Acremonium rutilum, under submerged fermentation. Curr Microbiol 58(2):122–128CrossRefGoogle Scholar
  20. 20.
    Raina A, Khatri R (2011) Quantitative determination of l-DOPA in seeds of Mucuna pruriens Germplasm by high performance thin layer chromatography. Indian J Pharm Sci 73(4):459–462Google Scholar
  21. 21.
    Sundaram U, Gurumoorthi P (2012) Validation of HPTLC method for quantitative estimation of l-DOPA from Mucuna pruriens. Int Res Pharm 3:300–304Google Scholar
  22. 22.
    Sampath VA, Novel MFK, Mani N, Babu UV (2013) A novel approach of the Isolation of l-DOPA from the methanolic extract of Mucuna pruriens seeds and its quantitative analysis by HPTLC. IJPPR 5(4):259–262Google Scholar
  23. 23.
    Shah PH, Joshi B (2010) Estimation of l-DOPA from Mucuna pruriens Linn and formulations containing M. pruriens by spectrofluorimetric method. Int J Pharmtech Res 2(2):1033–1036Google Scholar
  24. 24.
    Mennickent S, Nail M, Vega M, de Diego M (2007) Quantitative determination of l-DOPA in tablets by high performance thin layer chromatography. J Sep Sci 30(12):1893–1898CrossRefGoogle Scholar
  25. 25.
    Stankovič DM, Samphao A, Dojcinović B, Kalcher K (2016) Rapid electrochemical method for the determination of l-DOPA in extract from the seeds of Mucuna prurita. Acta Chim Slov 63(2):220–226CrossRefGoogle Scholar
  26. 26.
    Mehran SMM, Golshani B (2013) Simultaneous determination of levodopa and carbidopa from fava bean, green peas and green beans by high performance liquid gas chromatography. J Clin Diagn Res 7(6):1004–1007Google Scholar
  27. 27.
    Ingale PK (2003) l-DOPA bearing plants. Nat Prod Radiance 2(3):126–133Google Scholar
  28. 28.
    Patil R, Gholave A, Yadav S, Bapat V, Jadhav J (2015) Mucuna sanjappae aitawade et Yadav: a new species of Mucuna with promising yield of anti-Parkinson’s drug l-DOPA. Genet Resour Crop Evol 62(1):155–162CrossRefGoogle Scholar
  29. 29.
    Soares AR, Marchiosi R, de Cássia Rita, Siqueira-Soares Rogério Barbosa, de Lima Wanderley, dos Santos Dantas, Ferrarese-Filho Osvaldo (2014) The role of l-DOPA in plants. Plant Signal Behav 9(4):e28275CrossRefGoogle Scholar

Copyright information

© The National Academy of Sciences, India 2019

Authors and Affiliations

  • Ravishankar Patil
    • 1
    • 2
    • 4
  • Chetan Aware
    • 2
  • Swaroopsingh Gaikwad
    • 3
  • Manisha Rajebhosale
    • 2
  • Vishwas Bapat
    • 2
  • Shrirang Yadav
    • 3
  • Jyoti Jadhav
    • 1
    • 2
    Email author
  1. 1.Department of BiochemistryShivaji UniversityVidyanagar, KolhapurIndia
  2. 2.Department of BiotechnologyShivaji UniversityVidyanagar, KolhapurIndia
  3. 3.Department of BotanyShivaji UniversityVidyanagar, KolhapurIndia
  4. 4.Amity Institute of BiotechnologyAmity UniversityMumbaiIndia

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