Development of Analytical Screening Method for Rapid Identification of Carica Papaya Seed Adulterants in Black Pepper

Abstract

Adulteration is a critical issue that undermines the credibility and therapeutic potential of Ayurvedic medicine. Ensuring authenticity through scientific methods and regulatory oversight is essential to preserve the heritage and promote global acceptance of Ayurveda. With regards to above concern HPLC and HPTLC method has been established for identification of adulteration of Carica papaya seed in black pepper. The methanolic extracts of Carica papaya seeds, along with authenticated samples of Piper nigrum and an adulterated mixture, were simultaneously applied onto the TLC plate. After the development, the plates were scanned at a wavelength of 254 nm. The HPLC analysis showing specific Rt at 28.1 min. for the specific marker present in Carica papaya seeds and resulting chromatogram of HPTLC revealed a distinct Rf value of 0.69 in both the Carica papaya seed extract and the adulterated mixture track, thereby proving the presence of papaya seed in the adulterated sample. This method offers a reliable approach for rapid identification of Carica papaya seed adulteration in black pepper.

Keywords

HPTLC / adulteration / quality control / standardization / ayurveda / piperine

References

[1] Wu K.; Ruan C.Acupuncture for Chronic Obstructive Pulmonary Disease: A 38-Year Bibliometric Landscape of Global Research Trends and Knowledge Evolution ( 1986-2024). Int. J. Chron. Obstruct. Pulmon. Dis. 2025, 20, 2393-2408.

[2] Lepcha I.; Patra B.; Saha D.Medicinal Plants in Ayurveda and Unani Systems including Siddha in the Indian Scenario; Shashwat Publication: Bilaspur, India, 2023.

[3] Wang H.; Chen Y.; Wang L.; et al. Advancing herbal medicine: Enhancing product quality and safety through robust quality control practices. Front. Pharmacol. 2023, 14, 1265178.

[4] Ekar T.; Kreft S. Common risks of adulterated and mislabeled herbal preparations. Food Chem. Toxicol. 2019, 123, 288-297.

[5] Ichim M.C. The DNA-based authentication of commercial herbal products reveals their globally widespread adulteration. Front. Pharmacol. 2019, 10, 1227.

[6] Mohite P.; Puri A. Adulteration and evaluation of crude drugs of natural origin. In Pharmacognosy and Phytochemistry: Principles, Techniques, and Clinical Applications, 1st ed.; Willey: Hoboken, NJ, USA, 2025; pp. 101-121.

[7] Saggar S.; Mir P.A.; Kumar N.; et al. Traditional and herbal medicines: Opportunities and challenges. Pharmacog. Res. 2022, 14, 107-114.

[8] Balkrishna A.; Sharma N.; Srivastava D.; et al. Exploring the safety, efficacy, and bioactivity of herbal medicines: Bridging traditional wisdom and modern science in healthcare. Future Integr. Med. 2024, 3, 35-49.

[9] Acharya R.; Narayanan R.V.; Mahajon B.; et al. Contributions to research and development by Central Council for Research in Ayurvedic Sciences (CCRAS): An appraisal. Int. J. Res. 2022, 3, 86-101.

[10] Alswaidi F.M.; Abualssayl A.A. Ayurveda; safety, effectiveness, and acceptance around the world. F1000Research 2025, 14, 62.

[11] Verma S.K.; Pandey M.; Sharma A.; et al. Exploring Ayurveda: Principles and their application in modern medicine. Bull. Natl. Res. Centre 2024, 48, 77.

[12] Pugalenthi M.; Deepa M.A.; Pradheeba M.; et al. An Insight to Intensify the Potential Prospects of Underutilized Wild Species of Piper. In Pomotherapeutic Insights on Wild Edible Fruits; Springer Nature Singapore: Singapore, 2025; pp. 89-113.

[13] Patil P.; Gupta M.; Semwal A.; et al. Reviewing Bioenhancers in Traditional Indian Medicine: Exploring their Potential Mechanisms and Applications. Front. Health Infor. 2024, 13, 256.

[14] Haq I.U.; Imran M.; Nadeem M.; et al. Piperine: A review of its biological effects. Phytother. Res. 2021, 35, 680-700.

[15] Singh S.; Shukla A. Therapeutic potential of piperine: A comprehensive review. Nat. Prod. Jour. 2024, 14, 53-67.

[16] Variyar P.S.; Singh I.P.; Adiani V.; et al. Peppers:Biological, Health, and Postharvest Perspectives; CRC Press:Boca Raton, FL, USA, 2024.

[17] Ibrahim M.; Detroja A.; Sheth B.P.; et al. Existing status and future advancements of adulteration detection techniques in herbal products. Mol. Bio. Rep. 2024, 51, 151.

[18] Paul T.; Kumar K.J. Standardization of herbal medicines for lifestyle diseases. In Role of Herbal Medicines:Management of Lifestyle Diseases; Springer Nature: Singapore, 2024; pp. 545-557.

[19] Takooree H.; Aumeeruddy M.Z.; Rengasamy K.R.; et al. A systematic review on black pepper (Piper nigrum L.): From folk uses to pharmacological applications. Crit. Rev. Food Sci. Nutr. 2019, 59, S210-S243.

[20] Ayurvedic Pharmacopoeia Committee. The Ayurvedic Formulary of India; Controller of Publications: Delhi, India, 2003.

[21] Joshi V.K.; Joshi A.; Dhiman K.S. The Ayurvedic Pharmacopoeia of India, development and perspectives. J. Ethnopharm. 2017, 197, 32-38.

[22] The Ayurvedic Pharmacopoeia of India. Available online: http://www.ayurveda.hu/api/API-Vol-3.pdf (accessed on 16 May 2024).

[23] Department of Ayurveda.Thin Layer Chromatographic (TLC) Atlas of Ayurvedic Pharmacopoeial Drugs, 1st ed.; Part I, Volume I; Department of Ayurveda: New Delhi, India, 2016.

[24] Khan Z.R.; Moni F.; Sharmin S.; et al. Isolation of bulk amount of piperine as active pharmaceutical ingredient (API) from black pepper and white pepper (Piper nigrum L.). Pharmacol. Pharm. 2017, 8, 253-262.

[25] Parihar A.K.S.; Sahu U.; Karbhal K.S.; et al. Quality Control of Triphala Churna. J. Med. Nat. Prod. 2024, 1, 100003.

[26] Sahu U.; Chauhan N.S.; Parihar A.K.S.; et al. Development of Simultaneous HPTLC Method and Validation for the Quality Assessment of Ayurvedic Formulation—Ayush Kvatha Churna by Using Marker Compound Rosmarinic Acid, Trans-Cinnamaldehyde and Piperine. J. Chromatogr. Sci. 2025, 63, bmae019.

[27] Patel S.; Baghel G.S.; Chauhan N.S.; et al. High-performance thin-layer chromatographic standardization and quantification of marker compounds in an Ayurvedic polyherbal formulation: Krishnadi Churna. JPC--Modern TLC 2021, 34, 493-502.

[28] Parihar A.K.; Sahu U.; Karbhal K.S.; et al. Green HPTLC-method to estimate trans-Cinnamaldehyde in Ayurvedic formulation Sitopaladi Churna. TMMM 2023, 6, 1-8.