COMPREHENSIVE PHARMACOGNOSTIC AND PHYTOCHEMICAL PROFILING OF TABERNAEMONTANA ALTERNIFOLIA LEAVES: QUANTIFICATION OF PHENOLICS, ALKALOIDS, FLAVONOIDS, AND EVALUATION OF AFLATOXIN AND HEAVY METAL CONTAMINATION

Authors

  • PALLAVI V BHOKARE Reserach Scholar, Marthwada Mitra Mandal’s College of Pharmacy, Thergaon (Kalewadi), (Affiliated to Savitribai Phule Pune University, Pune) Pune, Maharashtra, India https://orcid.org/0000-0003-2154-4931
  • MANOHAR J PATIL Reserach Scholar, Marthwada Mitra Mandal’s College of Pharmacy, Thergaon (Kalewadi), (Affiliated to Savitribai Phule Pune University, Pune) Pune, Maharashtra, India
  • PARSAD V KADAM Reserach Scholar, Marthwada Mitra Mandal’s College of Pharmacy, Thergaon (Kalewadi), (Affiliated to Savitribai Phule Pune University, Pune) Pune, Maharashtra, India https://orcid.org/0000-0002-9599-2123

DOI:

https://doi.org/10.22159/ajpcr.2026v19i2.57614

Keywords:

Pharmacognosy, Phenolic content, Flavonoids, Alkaloids, Heavy metal analysis, Aflatoxins, Herbal standardisation

Abstract

Objectives: The study aimed to establish pharmacognostic standards and quantify major bioactive constituents, phenolics, alkaloids, and flavonoids in the leaves of Tabernaemontana alternifolia (Apocynaceae).

Methods: Leaves were collected from the Ratnagiri region of Maharashtra, India, authenticated, and subjected to detailed macroscopic, microscopic, physicochemical, and phytochemical evaluations. Microscopy was used to identify diagnostic anatomical features, whereas physicochemical parameters such as ash values and extractive values were determined using standard procedures. Qualitative and quantitative phytochemical analyses were performed to assess the presence and levels of key metabolites. Heavy metal content was analyzed using atomic absorption spectroscopy, and aflatoxins were assessed using liquid chromatography-tandem mass spectrometry (MS)/MS.

Results: Microscopic examination revealed a dorsiventral leaf with bicollateral vascular bundles and abundant microsphenoidal calcium oxalate crystals. Physicochemical parameters included total ash (5.67±0.76% w/w), acid-insoluble ash (0.54±0.01% w/w), and water-soluble ash (3.21±0.04% w/w), confirming minimal inorganic contamination. Soxhlet extraction produced the highest extractive value (17.93±1.37% w/w). Phytochemical screening confirmed the presence of alkaloids, phenolics, flavonoids, tannins, saponins, proteins, and reducing sugars. Quantitative analysis showed notable levels of total phenolics (11.82 mg gallic acid equivalents/g), flavonoids (7.53 mg rutin equivalents/g), and alkaloids (4.0 mg atropine equivalents/g). Heavy metals (Cd, Pb, As, and Hg) were below detectable limits, and aflatoxins (B1, B2, G1, and G2) were absent.

Conclusion: This study establishes key pharmacognostic and phytochemical benchmarks for T. alternifolia leaves. Soxhlet extraction yielded the highest extractive value, whereas quantitative analyses revealed appreciable levels of phenolics, flavonoids, and alkaloids, indicating notable bioactive potential. The absence of heavy metals and aflatoxins confirms material safety, supporting its standardization, quality control, and further pharmacological development.

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References

1. Gonçalves BM, Duarte N, Ramalhete C, Barbosa F, Madureira AM, Ferreira MJ. Monoterpene indole alkaloids with anticancer activity from Tabernaemontana species. Phytochem Rev. 2025 Jun 27;24(3):2271-307. doi: 10.1007/s11101-024-09964-6

2. Neela S, Ajitha M, Kuchana V. Comprehensive phytochemical profiling, GC-MS analysis, molecular docking and antiproliferative activity of ethanol fraction of Tabernaemontana coronaria and Thunbergia alata. J Nat Remedies. 2024 Aug 31;24:1739-51. doi: 10.18311/jnr/2024/41547

3. Jain D, Meena M, Janmeda P, Seth CS, Arora J. Analysis of quantitative phytochemical content and antioxidant activity of leaf, stem, and bark of Gymnosporia senegalensis (Lam.) Loes. Plants (Basel). 2024 May 21;13(11):1425. doi: 10.3390/plants13111425, PMID 38891234

4. Das S, Sultana KW, Mondal M, Chandra I, Ndhlala AR. Unveiling the dual nature of heavy metals: Stressors and promoters of phenolic compound biosynthesis in Basilicum polystachyon (L.) Moench in vitro. Plants (Basel). 2023 Dec 28;13(1):98. doi: 10.3390/plants13010098, PMID 38202406

5. Validandi V, Kurella S, Gorain S, Sagubandi Y, Rudhra A, Annamanedi SK, et al. Heavy metal contamination in commonly consumed herbal medicines from local markets in Hyderabad, India, and associated health risks. Food Addit Contam Part B Surveill. 2025 Apr 3;18(2):187-98. doi: 10.1080/19393210.2025.2471065, PMID 40009029

6. Rani S, Sisodia R. Heavy metal contamination of medicinal plants in India - A perspective. Asian J Water Environ Pollut. 2023 Nov 27;20(6):9-17. doi: 10.3233/ajw230074

7. Khatoon A, Jain S, Shukla K, Bapna RS. Phytochemical investigation and determination of total phenols, flavonoid, saponins and alkaloids concentration in root extract of Tabernaemontana divaricata R.Br. Ex roem. And schult. J Pharm Negat Results. 2022;13(10):4822-6.

8. Islam MM, Saxena N, Sharma D. Phytoremediation as a green and sustainable prospective method for heavy metal contamination: A review. RSC Sustain. 2024;2(5):1269-88. doi: 10.1039/d3su00440f

9. Naidoo CM, Naidoo Y, Dewir YH, Singh M, Daniels AN, El-Ramady H. In vitro investigation of the antioxidant and cytotoxic potential of Tabernaemontana ventricosa Hochst. Ex A. DC. Leaf, stem, and latex extracts. Horticulturae. 2022 Jan 20;8(2):91. doi: 10.3390/ horticulturae8020091

10. Sari R, Conterno P, Da Silva LD, De Lima VA, Oldoni TL, Thomé GR, et al. Extraction of phenolic compounds from Tabernaemontana catharinensis leaves and their effect on oxidative stress markers in diabetic rats. Molecules. 2020 May 21;25(10):2391. doi: 10.3390/ molecules25102391, PMID 32455579

11. Naidoo CM, Naidoo Y, Dewir YH, Murthy HN, El-Hendawy S, Al-Suhaibani N. Major bioactive alkaloids and biological activities of Tabernaemontana species (Apocynaceae). Plants (Basel). 2021 Feb 5;10(2):313. doi: 10.3390/plants10020313, PMID 33562893

12. Kawade M, Ingole V, Shinde A, Katade S. Qualitative phytochemical screening and metal analysis of some selected medicinal plants. S East Eur J Public Health. 2025 May 1;27:1313-7. doi: 10.70135/seejph. vi.6394

13. Alimuddin AH, Rudiyansyah R, Masriani M. Penetapan Kadar flavonoid, fenolik dan aktivitas antioksidan ekstrak buah Tabernaemontana macrocarpa Jack Asal Kalimantan Barat (Determination of flavonoid, phenolic, and antioxidant activity of Tabernaemontana macrocarpa jack fruit extract from west Kalimantan). Indones J Pure Appl Chem. 2023 Oct 31;6(3):132-41. doi: 10.26418/indonesian.v6i3.63749

14. Trease GE, Evans WC. Textbook of Pharmacognosy. 5th ed. St Louis, MO: Elsevier; 2009.

15. Adeniran AA, Ntamanwuna EC, Bassey VO. Microscopical characterization and physicochemical standardization of leaves, stems and roots of Spondias mombin L. (Anacardiaceae). Niger J Pharm Res. 2021 Aug 12;17(1):15-25. doi: 10.4314/njpr.v17i1.2

16. Nugraha SE, Keliat JM, Marianne, Syahputra RA. Investigating the toxicity of betalain compounds: In silico analysis and in vivo predictions for standardized Beta vulgaris L. Extract. Int J Appl Pharm. 2024 Jan 7;16:118-23. doi: 10.22159/ijap.2024v16i1.49189

17. Panchal CV, Sawale JA, Poula BN, Khandelwal KR. Pharmacognostic studies of Lagenaria siceraria (Molina) Standley fruits. Pharmacogn J. 2014 Feb 20;6(1):7-11. doi: 10.5530/pj.2014.1.2

18. Chakraborty S, Pal A. Bala NN. Comparative study of yields and total phenolic and flavonoid contents of extracts of different extraction methods of Clitoria ternatea (Fabaceae). Int J Curr Pharm Res. 2022;14(6):32-5.

19. Muhaimin M, Latifah N, Chaerunisaa AY, Amalia E, Rostinawati T. Preparation and characterization of Sonneratia alba leaf extract microcapsules by solvent evaporation technique. Int J Appl Pharm. 2022 Nov 7;14:77-82. doi: 10.22159/ijap.2022v14i6.46274

20. Othman NA, Idris SA, Rosli NR. Maceration and soxhlet extraction of Orthosiphon stamineus - a comparative study. AIP Conf Proc. 2024;3041:050007. doi: 10.1063/5.0195348

21. Wu EY, Sun WJ, Wang Y, Zhang GY, Xu BC, Chen XG, et al. Optimization of ultrasonic-assisted extraction of total flavonoids from Abrus cantoniensis (Abriherba) by response surface methodology and evaluation of its anti-inflammatory effect. Molecules. 2022 Mar 22;27(7):2036. doi: 10.3390/molecules27072036, PMID 35408435

22. Azad N, Chhatria C. Phytochemical profiling of Asteracantha longifolia and Moringa Oleifera: A comparative analysis of Soxhlet and microwave-assisted extraction method. Int J Pharm Sci Res. 2025;16(10):2820-8.

23. Pourmorad F, Hosseinimehr SJ Shahabimajd N. Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants. Afr J Biotechnol. 2006;5(11):1142-5.

24. Mwamatope B, Tembo D, Kampira E, Maliwichi-Nyirenda C, Ndolo V. Seasonal variation of phytochemicals in four selected medicinal plants. Pharmacogn Res. 2021 Oct 20;13(4):218-26. doi: 10.5530/pres.13.4.14

25. Mwamatope B, Tembo D, Chikowe I, Kampira E, Nyirenda C. Total phenolic contents and antioxidant activity of Senna singueana, Melia azedarach, Moringa oleifera and Lannea discolor herbal plants. Sci Afr. 2020 Sep;9:e00481. doi: 10.1016/j.sciaf.2020.e00481

26. Puro KN, Mazumder MU, Khazeo P, Jyrwa R, Jamir N, Sailo L. Qualitative and Quantitative Analysis of Phytochemicals of Crude Extracts of Ageratum conyzoides L. Leaves. In: Proceedings of the Mizoram Science Congress 2018 (MSC 2018) - Perspective and Trends in the Development of Science Education and Research. Paris, France: Atlantis Press; 2018. Available from: https://www.atlantis-press.com/ article/55909747

27. Mukhopadhyay G. Atomic spectroscopy analysis of heavy metals in plants. Int J Pharm Biol Arch. 2018;9(3):1-5.

28. Liu L, Jin H, Sun L, Ma S, Lin R. Determination of aflatoxins in medicinal herbs by high‐performance liquid chromatography-tandem mass spectrometry. Phytochem Anal. 2012 Sep 9;23(5):469-76. doi: 10.1002/pca.2343, PMID 22323053

29. Mishevski P. Ivanovska TP, Zhivikj Z, Petrushevska-Tozi L, Najdenkoska A, Mincheva M. LC-MS/MS method for detecting trace levels of aflatoxin B1 in cultivated cannabis from Republic of North Macedonia. Maced Pharm Bull. 2024;70(2):33-9.

30. Sonibare MA, Isola AO, Akinmurele OJ. Pharmacognostic standardisation of the leaves of Costus afer Ker Gawl. (Zingiberaceae) and Palisota hirsuta (Thunb.) K. Schum. (Commelinaceae). Futur J Pharm Sci. 2023 Mar 6;9(1):19. doi: 10.1186/s43094-023-00469-1

31. Karanje P, Ghorpade V, Namdeo A, Doijad R, Karanje A. Comparison of techniques for the extraction of camptothecin from Tabernaemontana species. Nat Prod Res. 2023 Jan 17;37(2):322-7. doi: 10.1080/14786419.2021.1957886, PMID 34319175

32. Manna P, Jain SK. Obesity, oxidative stress, adipose tissue dysfunction, and the associated health risks: Causes and therapeutic strategies. Metab Syndr Relat Disord. 2015 Dec;13(10):423-44. doi: 10.1089/ met.2015.0095, PMID 26569333

33. Zhang YJ, Gan RY, Li S, Zhou Y, Li AN, Xu DP, et al. Antioxidant phytochemicals for the prevention and treatment of chronic diseases. Molecules. 2015 Nov 27;20(12):21138-56. doi: 10.3390/ molecules201219753, PMID 26633317

34. Sathishkumar T, Baskar R, Aravind M, Tilak S, Deepthi S, Bharathikumar VM. Simultaneous extraction optimization and analysis of flavonoids from the flowers of Tabernaemontana heyneana by high performance liquid chromatography coupled to diode array detector and electron spray ionization/mass spectrometry. ISRN Biotechnol. 2013;2013:450948. doi: 10.5402/2013/450948, PMID 25969771

35. Mondal S, Chakrabarty K, Bose M, Maity S, Chowdhury SS, Das S, et al. Exploring the therapeutic potential: In-vitro assessment of antioxidant and anti-inflammatory activity of methanolic extract from Tabernaemontana divaricata leaves. J Surv Fish Sci. 2024 Mar 5;11:134-40. doi: 10.53555/sfs.v11i3.2405

36. Tadkase PP, Ghiware NB, Shaikh H, Vidhya V, Mali AV. Comparative evaluation of Tabernaemontana divaricata leaves extracts for antidepressant activity. World J Pharm Res. 2018;7(12):641-50.

Published

07-02-2026

How to Cite

PALLAVI V BHOKARE, et al. “COMPREHENSIVE PHARMACOGNOSTIC AND PHYTOCHEMICAL PROFILING OF TABERNAEMONTANA ALTERNIFOLIA LEAVES: QUANTIFICATION OF PHENOLICS, ALKALOIDS, FLAVONOIDS, AND EVALUATION OF AFLATOXIN AND HEAVY METAL CONTAMINATION”. Asian Journal of Pharmaceutical and Clinical Research, vol. 19, no. 2, Feb. 2026, pp. 144-52, doi:10.22159/ajpcr.2026v19i2.57614.

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Original Article(s)