Comparative Study to Assess Chemo-Profile variation of Maricha (piper nigrum Linn) between Non-Grafted and Grafted Variety Sample by RP-UPLC Method

Abstract

In Ayurveda, therapeutic success depends on four essential factors, among which Dravya (medicine) holds prime importance after the physician. Traditionally, medicinal plants were collected directly by Vaidyas; however, current practices largely depend on market-sourced raw materials. Maricha (Piper nigrum Linn.) is an extensively used drug in Ayurvedic formulations, and due to increasing demand, modern cultivation techniques such as grafting are being adopted over conventional methods. Grafting is known to enhance yield, improve disease resistance, and reduce harvesting time. The present study aims to compare the pharmacognostic, physicochemical, phytochemical, and chemical profiles of non-grafted and grafted Maricha. Fresh seeds of Piper nigrum cultivated through non-grafted and grafted methods were collected and shade-dried following Good Cultivation Practices (GCP). Both samples were evaluated through pharmacognostic, physicochemical, and phytochemical analyses, along with chromatographic profiling using the RP-UPLC method. The results revealed that grafted plants exhibited superior morphological characteristics, including thicker stems, larger leaves, and bigger fruits. Phytochemical screening showed the presence of similar constituents such as saponins, tannins, alkaloids, volatile oils, and starch in both samples, while physicochemical parameters remained within standard limits. RP-UPLC analysis indicated a higher piperine content in grafted Maricha (4.57%) compared to non-grafted samples (3.57%). The study concludes that grafting positively influences the morphological and chemical attributes of Maricha, supporting its use for improved quality and efficacy of Ayurvedic formulations and emphasizing the need for further research on genetic and environmental influences.

Introduction

In Ayurveda, Dravya (medicinal substance) is a fundamental component of treatment under Chikitsa Chatushpada, and its therapeutic effectiveness depends not only on inherent properties but also on proper cultivation, collection, processing, and storage. With modern commercial sourcing of raw materials, concerns regarding quality and authenticity have increased. Scientific cultivation practices are therefore essential to ensure safety, efficacy, sustainability, and phytochemical integrity of medicinal plants.

Maricha (Piper nigrum Linn., black pepper) is a widely used Ayurvedic drug and key ingredient in classical formulations such as Trikatu. Due to high demand, cultivation methods have evolved, including the adoption of grafting techniques, where a scion is grafted onto disease-resistant rootstock (e.g., Piper colubrinum) to improve yield and resistance to foot rot disease. Although grafting offers agronomic benefits, its impact on the chemical composition and medicinal properties of Maricha remains unclear, necessitating scientific evaluation.

The study aims to compare the chemoprofile variations between grafted and non-grafted Maricha. Objectives include pharmacognostic, physicochemical, and phytochemical analysis, along with quantitative estimation of piperine (the major active constituent) using RP-UPLC.

Methodology involved collecting grafted (M120) and non-grafted (M121) samples from the same field to control environmental variables. After authentication, samples were dried, powdered, and subjected to:

• Pharmacognostic evaluation (macroscopic and microscopic analysis)

• Physicochemical tests (moisture content, ash values, extractive values, pH, foreign matter)

• Phytochemical screening (organic and inorganic constituents)

• TLC analysis using piperine as a reference standard

• RP-UPLC analysis for quantitative estimation of piperine under standardized chromatographic conditions

A calibration curve for piperine (100–500 µg/mL) was prepared to ensure accurate quantification.

Overall, the study seeks to scientifically assess whether grafting influences the phytochemical composition and therapeutic quality of Maricha, thereby supporting evidence-based cultivation and standardization in Ayurvedic medicine.

Conclusion

1) Macroscopic study reveals better growth of the Grafted Maricha variety compared to Non-grafted Maricha variety in size, shape and plant growth. 2) Organoleptic study shows presence of higher concentrations of aromatic compounds in Non-grafted Maricha sample compared to Grafted Maricha sample. 3) Microscopic study of both non-grafted and grafted samples confirmed that grafting didn’t alter internal anatomy of Maricha. 4) Physicochemical and Phytochemical studies showed the results as per API standards in both the samples thus confirming genuinity. 5) TLC results confirmed additional bioactive components in the grafted variety of Maricha sample. 6) RP-UPLC study carried out for quantification of Piperine in both the samples confirmed the genuinity and higher Chemo-profile was observed in Grafted variety of Maricha. 7) Grafting was effective not only for increasing yield, Reduction of Harvest period and Foot rot infestation in cultivation of Maricha but also have additional chemo-profile variation compared to conventional method of cultivation with quantitative increase in important bioactive compounds such as Piperine. 8) Thus, Grafting may be beneficial in additional medicinal properties compared to conventional method of cultivation.

References

[1] Agnivesha. Charaka Samhita. Elaborated by Charaka and Dridhabala. Edited with Ayurveda Deepika sanskrit commentary by Vaidya Jhadavji Trikamji. Varanasi: Chaukhamba Orientalia; 2015. p. 23. Verse 126. [2] Bhavapraksha. Bhavaprakasha Nighantu. Edited with Hindi vyakhyana by Padmshri Prof. Chunekar Krishnachandra. Varanasi:Chaukamba Bharathi Academy;2020. p.18. Verse 62-63. [3] Bhavapraksha. Bhavaprakasha Nighantu. Edited with Hindi vyakhyana by Padmshri Prof. Chunekar Krishnachandra. Varanasi:Chaukamba Bharathi Academy;2020. p.19. Verse 66. [4] Bhavapraksha. Bhavaprakasha Nighantu. Edited with Hindi vyakhyana by Padmshri Prof. Chunekar Krishnachandra. Varanasi:Chaukamba Bharathi Academy;2020. p.19. Verse 66. [5] Nysanth NS, Divya S, Nair CB, Anju AB, Praveena R, Anith KN. Biological control of foot rot (Phytophthora capsici Leonian) disease in black pepper (Piper nigrum L.) with rhizospheric microorganisms. Rhizosphere. 2022;23:100578. doi:10.1016/j.rhisph.2022.100578. [6] Council of Scientific and Industrial Research. The Wealth of India: A Dictionary of Indian Raw Materials and Industrial Products. 1st ed. Raw Materials, Vol. 8. New Delhi: Council of Scientific and Industrial Research; p. 111. [7] Council of Scientific and Industrial Research. The Wealth of India: A Dictionary of Indian Raw Materials and Industrial Products. 1st ed. Raw Materials, Vol. 8. New Delhi: Council of Scientific and Industrial Research; p. 109. [8] Ministry of Health and Family Welfare, Govt of India. Ayurvedic Pharmacopoeia of India. 1st ed. Reprint 2001. Part 1, Vol III. New Delhi: Dept of Indian Systems of Medicine and Homeopathy, Controller of Publications Civil Lines; 2001. p.115. [9] Ministry of Health and Family Welfare, Govt of India. Ayurvedic Pharmacopoeia of India. 1st ed. Reprint 2001. Part 1, Vol III. New Delhi: Dept of Indian Systems of Medicine and Homeopathy, Controller of Publications Civil Lines; 2001. p.115. [10] Sushruta. Sushruta Samhita. Edited with Nibandhasangraha commentary of Sri Dalhanacharya by Vaidya Jhadavji Trikamji. Varanasi: Chaukhambha Surbharati prakashan; 2003.Chikitsa sthana. Verse 28/27

Copyright

Copyright © 2026 Dr. Guruprasad Hegde, Dr. Rajashekar S Ganigera, Dr. Dharani , Dr. Shivananda B Karigar, Dr. Sampathkumar Bellamma. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.