Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract

Poro David Clark; Erhuvwu Mercy Osharode; Johnson Oji Otutu; Kehinde Johnathan Awatefe; Avwoghokoghene Christopher Idibie; Violet Oshenyen Ejomafuvwe

doi:10.21448/ijsm.1795990

Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract

Abstract

This research investigated the phytochemical components and antioxidant properties of the ethanol leaves extract fraction (C5), which was obtained by macerating the leaves in 80% ethanol, fractionating the extract via column chromatography, and further purifying it using TLC. The resulting purified fraction was characterized through UV–Vis, FTIR, and GC–MS analyses. Qualitative phytochemical screening of fraction C5 confirmed the presence of flavonoids, alkaloids, terpenoids, saponins, and cardiac glycosides. UV-Vis spectroscopy revealed an absorption maximum at 406 nm, suggesting extended conjugated systems such as oxidized polyphenolics or carotenoid derivatives. FTIR analysis showed characteristic bands, including O-H stretching at 3200–3600 cm⁻¹ (indicative of phenolic and hydroxyl groups), aromatic C=C stretching at 1450–1600 cm⁻¹, aliphatic C-H stretching at 2850–2950 cm⁻¹, and C-O stretching at 1000–1300 cm⁻¹ (related to ethers, alcohols, or ester linkages). GC-MS profiling identified 14 constituents, including terpenoids, methoxylated phenylpropanoids, phytosterols, fatty acids, and the therapeutically significant indole alkaloid reserpine. In-vitro antioxidant assays showed clear, concentration dependent activity in all three methods: at 2.0 mg/mL, fraction C5 produced 71.4% DPPH inhibition, 76.8% ABTS inhibition, and 72.5% ferric reducing activity, whereas gallic acid was significantly more potent at every concentration (p < 0.001), both exhibited a similar linear dose–response. These findings offer scientific support for the traditional medicinal use of Rauvolfia vomitoria Afzel. leaves and highlight its potential as a natural source of antioxidants for managing oxidative stress-related conditions.

Keywords

Supporting Institution

Edwin Clark University

Project Number

nil

Ethical Statement

This study involved the collection and analysis of plant material (Rauvolfia vomitoria leaves) and did not require ethical approval as no human subjects, animal subjects, or endangered species were involved. Plant samples were collected following sustainable harvesting practices to minimize environmental impact, and all laboratory analyses were conducted in accordance with standard safety protocols and institutional guidelines.

Thanks

The authors are grateful to their respective institutions for providing laboratory facilities and technical support. Special thanks are due to the technical staff of Edwin Clark University for their assistance during the experimental work.

References

Abubakar, A.R., & Haque, M. (2020). Preparation of medicinal plants: Basic extraction and fractionation procedures for experimental purposes. Journal of Pharmacy & Bioallied Sciences, 12(1), 1–10. https://doi.org/10.4103/jpbs.JPBS_175_19
Ajayi, O.A. (2021). Phytochemical and GC-MS analysis of bioactive components in ethanolic extract of Rauvolfia vomitoria leaves. Journal of the Chemical Society of Nigeria, 46(4), 656–660. https://doi.org/10.46602/jcsn.v46i4.642
Arya, O.P., Bhatt, I.D., & Mohanty, K. (2025). Improving phenolic antioxidants in Illicium griffithii fruit through optimizing subcritical water extraction method. Scientific Reports, 15, 37387. https://doi.org/10.1038/s41598-025-16989-0
Asoro, I.I., Ebuehi, O.A.T., & Igwo-Ezikpe, M.N. (2021). GC-MS analysis of the Rauvolfia vomitoria ethanol extracts. European Journal of Medicinal Plants, 32(6), 34 45. https://doi.org/10.9734/ejmp/2021/v32i630398
Baliyan, S., Mukherjee, R., Priyadarshini, A., Vibhuti, A., Gupta, A., Pandey, R.P., & Chang, C.-M. (2022). Determination of antioxidants by DPPH radical scavenging activity and quantitative phytochemical analysis of Ficus religiosa. Molecules, 27(4), 1326. https://doi.org/10.3390/molecules27041326
Baskar, A.A., Al Numair, K.S., Paulraj, M.G., Alsaif, M.A., Al Muamar, M., & Ignacimuthu, S. (2012). β-Sitosterol prevents lipid peroxidation and improves antioxidant status and histoarchitecture in rats with 1,2-dimethylhydrazine-induced colon cancer. Journal of Medicinal Food, 15(4), 335–343. https://doi.org/10.1089/jmf.2011.1780
Chaves, N., Santiago, A., & Alías, J.C. (2020). Quantification of the antioxidant activity of plant extracts: Analysis of sensitivity and hierarchization based on the method used. Antioxidants, 9(1), 76. https://doi.org/10.3390/antiox9010076
Clark, P.D., & Ekpekpo, L.D. (2025). Maceration-based extraction and spectroscopic characterisation of Sorghum bicolor leaves extract as a natural dye on cotton fabric. Journal of the Chemical Society of Nigeria, 50(4), 775–786. https://doi.org/10.4314/jcsn.v50i4.7

Clark, P.D., & Omo-Udoyo, E. (2021). A comparative assessment on antioxidant and phytochemical of Trichilia monadelpha (Thonn) J.J. De Wilde (Meliaceae) plant extracts. Chemical Science International Journal, 30(10), 24–33. https://doi.org/10.9734/CSJI/2021/v30i1030257
Clark, P.D., Ndukwe, G.I., & Awatefe, K.J. (2026). Phytochemical characterization and evaluation of antioxidant activity in Sorghum bicolor leaves extracts. Sciences of Phytochemistry, 5(1), 31–42. https://doi.org/10.58920/sciphy0501459
El Mannoubi, I. (2023). Impact of different solvents on extraction yield, phenolic composition, in vitro antioxidant and antibacterial activities of deseeded Opuntia stricta fruit. Journal of Umm Al-Qura University for Applied Sciences, 9, 176–184. https://doi.org/10.1007/s43994-023-00031-y
Ilyasov, I.R., Beloborodov, V.L., Selivanova, I.A., & Terekhov, R.P. (2020). ABTS/potassium persulfate decolorization assay of antioxidant capacity: Mechanisms and main influencing factors. International Journal of Molecular Sciences, 21(3), 1131. https://doi.org/10.3390/ijms21031131
Iqbal, M.J., Kabeer, A., Abbas, Z., Siddiqui, H.A., Calina, D., Sharifi-Rad, J., & Cho, W. C. (2024). Interplay of oxidative stress, cellular communication and signaling pathways in cancer. Cell Communication and Signaling, 22(1), Article 7. https://doi.org/10.1186/s12964-023-01398-5
Iwu, I.C., Chinonye, C.C., Iwuji, C.S., Obiagwu, I., Onu, U.L., Ukaoma, A.A., Chijioke-Okere, M., & Ezekoye, M.O. (2021). Chemical and medicinal properties of Rauvolfia vomitoria (Afzel) harvested from the south eastern Nigeria. Asian Journal of Chemical Sciences, 10(4), 56–71. https://doi.org/10.9734/ajocs/2021/v10i419103
Jack, I.R., Clark, P.D., & Ndukwe, G.I. (2020). Evaluation of phytochemical, antimicrobial and antioxidant capacities of Pennisetum purpureum (Schumach) extracts. Chemical Science International Journal, 29(4), 1–14. https://doi.org/10.9734/csji/2020/v29i430170
Johnson, J., Mani, J., Ashwath, N., & Naiker, M. (2020). Potential for Fourier transform infrared (FTIR) spectroscopy toward predicting antioxidant and phenolic contents in powdered plant matrices. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 233, 118228. https://doi.org/10.1016/j.saa.2020.118228
Lee, J.-E., Jayakody, J.T.M., Kim, J.-I., Jeong, J.-W., Kim, T.-S., Seo, C., Azimi, I., Hyun, J., & Ryu, B. (2024). The influence of solvent choice on the extraction of bioactive compounds from Asteraceae: A comparative review. Foods, 13, 3151. https://doi.org/10.3390/foods13193151
Lezoul, N.E.H., Belkadi, M., Habibi, F., & Guillén, F. (2020). Extraction processes with several solvents on total bioactive compounds in different organs of three medicinal plants. Molecules, 25(20), 4672. https://doi.org/10.3390/molecules25204672
Ndukwe, G.I., Clark, P.D., & Jack, I.R. (2020). In vitro antioxidant and antimicrobial potentials of three extracts of Amaranthus hybridus L. leaf and their phytochemicals. European Chemical Bulletin, 9(7), 164–173. https://doi.org/10.17628/ecb.2020.9.164-173
Ndukwe, G.I., Tetam, J.G., & Clark, P.D. (2020). Phytochemical screening and in vitro antioxidant assessment of Cassia alata (Linn) leaf extracts. ChemSearch Journal, 11(2), 64–72.
Oborirhovo, O., Anigboro, A.A., Avwioroko, O.J., Akeghware, O., Okafor, B.J., Ovowa, F.O., & Tonukari, N.J. (2023). GC-MS characterized bioactive constituents and antioxidant capacities of aqueous and ethanolic leaf extracts of Rauvolfia vomitoria: A comparative study. Nigerian Journal of Science and Environment, 21(2), 479–499.
Onyeukwu, O.B., Achuba, F.I., & Okoro, I.O. (2025). Phytochemicals and gas chromatography–mass spectrometry (GC-MS) analysis of Rauvolfia vomitoria Afzel cold and hot water leaves extract. FUDMA Journal of Sciences, 9(3), 366–372. https://doi.org/10.33003/fjs-2025-0903-3125
Ouandaogo, H.S., Diallo, S., Odari, E., & Kinyua, J. (2023). Phytochemical screening and GC-MS analysis of methanolic and aqueous extracts of Ocimum kilimandscharicum leaves. ACS Omega, 8(50), 45738–45747. https://doi.org/10.1021/acsomega.3c07493
Pizzino, G., Irrera, N., Cucinotta, M., Pallio, G., Mannino, F., Arcoraci, V., Squadrito, F., Altavilla, D., & Bitto, A. (2017). Lifestyle, oxidative stress, and antioxidants: Back and forth in the pathophysiology of chronic diseases. Oxidative Medicine and Cellular Longevity, 2017, Article 8416763. https://doi.org/10.1155/2017/8416763
Rajurkar, N.S., & Hande, S.M. (2011). Estimation of phytochemical content and antioxidant activity of some selected traditional Indian medicinal plants. Indian Journal of Pharmaceutical Sciences, 73(2), 146–151. https://doi.org/10.4103/0250-474X.91574
Ramu, A.K., Ali, D., Alarifi, S., Abuthakir, M.H.S., & Abdul, B.A.A. (2021). Reserpine inhibits DNA repair, cell proliferation, invasion and induces apoptosis in oral carcinogenesis via modulation of TGF-β signaling. Life Sciences, 264, Article 118730. https://doi.org/10.1016/j.lfs.2020.118730
Syafriana, V., Azzahra, H.A., Vasya, Z.I., Subaryanti, & Muti, A.F. (2025). Effect of solvent type on the phytochemical content and the specific and non-specific parameters of Jombang (Taraxacum officinale) leaf extracts. Berita Biologi, 24(1), 149 161. https://doi.org/10.55981/berita_biologi.2025.8979
Tourabi, M., Faiz, K., Ezzouggari, R., Louasté, B., Merzouki, M., Dauelbait, M., Bourhia, M., Almaary, K.S., Siddique, F., Lyoussi, B., & Derwich, E. (2025). Optimization of extraction process and solvent polarities to enhance the recovery of phytochemical compounds, nutritional content, and biofunctional properties of Mentha longifolia L. extracts. Bioresources and Bioprocessing, 12, 76. https://doi.org/10.1186/s40643-025-00910-8
Ugwu, O.P.C., Okere, M., & Okon, M.B. (2022). Phytochemical analysis of ethanol leaf extract of Rauvolfia vomitoria. IAA Journal of Scientific Research, 8(1), 64–72.
Vittaya, L., Chalad, C., Ratsameepakai, W., & Leesakul, N. (2023). Phytochemical characterization of bioactive compounds extracted with different solvents from Calophyllum inophyllum flowers and activity against pathogenic bacteria. South African Journal of Botany, 154, 346–355. https://doi.org/10.1016/j.sajb.2023.01.052
Wongsa, P., Phatikulrungsun, P., & Prathumthong, S. (2022). FT-IR characteristics, phenolic profiles and inhibitory potential against digestive enzymes of 25 herbal infusions. Scientific Reports, 12, 6631. https://doi.org/10.1038/s41598-022-10669-z
Zeng, J., Zhang, D.-B., & Zhou, P.-P. (2017). Rauvomines A and B, two monoterpenoid indole alkaloids from Rauvolfia vomitoria. Organic Letters, 19(15), 4002 4005. https://doi.org/10.1021/acs.orglett.7b01723

Details

Primary Language

English

Subjects

Natural Products and Bioactive Compounds

Journal Section

Research Article

Authors

Poro David Clark ^*
0000-0001-7794-9642
Nigeria

Erhuvwu Mercy Osharode This is me
0009-0008-0089-7061
Nigeria

Johnson Oji Otutu
0000-0003-4544-7212
Nigeria

Kehinde Johnathan Awatefe This is me
0009-0001-5376-4237
Nigeria

Avwoghokoghene Christopher Idibie This is me
0009-0001-5135-922X
Nigeria

Violet Oshenyen Ejomafuvwe This is me
0009-0001-6704-3625
Nigeria

Early Pub Date

August 3, 2026

Publication Date

September 2, 2026

Submission Date

October 2, 2025

Acceptance Date

May 18, 2026

Published in Issue

Year 2026 Volume: 13 Number: 3

DOI

https://doi.org/10.21448/ijsm.1795990

IZ

https://izlik.org/JA43PF98FK

Cite

RIS / Bibtex

APA

Clark, P. D., Osharode, E. M., Otutu, J. O., Awatefe, K. J., Idibie, A. C., & Ejomafuvwe, V. O. (2026). Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract. International Journal of Secondary Metabolite, 13(3), 844-860. https://doi.org/10.21448/ijsm.1795990

AMA

1.Clark PD, Osharode EM, Otutu JO, Awatefe KJ, Idibie AC, Ejomafuvwe VO. Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract. Int. J. Sec. Metabolite. 2026;13(3):844-860. doi:10.21448/ijsm.1795990

Chicago

Clark, Poro David, Erhuvwu Mercy Osharode, Johnson Oji Otutu, Kehinde Johnathan Awatefe, Avwoghokoghene Christopher Idibie, and Violet Oshenyen Ejomafuvwe. 2026. “Multi-Instrumental Analysis of Bioactive Compounds and Antioxidant Properties in Rauvolfia Vomitoria Afzel. Leaves Extract”. International Journal of Secondary Metabolite 13 (3): 844-60. https://doi.org/10.21448/ijsm.1795990.

EndNote

Clark PD, Osharode EM, Otutu JO, Awatefe KJ, Idibie AC, Ejomafuvwe VO (September 1, 2026) Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract. International Journal of Secondary Metabolite 13 3 844–860.

IEEE

[1]P. D. Clark, E. M. Osharode, J. O. Otutu, K. J. Awatefe, A. C. Idibie, and V. O. Ejomafuvwe, “Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract”, Int. J. Sec. Metabolite, vol. 13, no. 3, pp. 844–860, Sept. 2026, doi: 10.21448/ijsm.1795990.

ISNAD

Clark, Poro David - Osharode, Erhuvwu Mercy - Otutu, Johnson Oji - Awatefe, Kehinde Johnathan - Idibie, Avwoghokoghene Christopher - Ejomafuvwe, Violet Oshenyen. “Multi-Instrumental Analysis of Bioactive Compounds and Antioxidant Properties in Rauvolfia Vomitoria Afzel. Leaves Extract”. International Journal of Secondary Metabolite 13/3 (September 1, 2026): 844-860. https://doi.org/10.21448/ijsm.1795990.

JAMA

1.Clark PD, Osharode EM, Otutu JO, Awatefe KJ, Idibie AC, Ejomafuvwe VO. Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract. Int. J. Sec. Metabolite. 2026;13:844–860.

MLA

Clark, Poro David, et al. “Multi-Instrumental Analysis of Bioactive Compounds and Antioxidant Properties in Rauvolfia Vomitoria Afzel. Leaves Extract”. International Journal of Secondary Metabolite, vol. 13, no. 3, Sept. 2026, pp. 844-60, doi:10.21448/ijsm.1795990.

Vancouver

1.Poro David Clark, Erhuvwu Mercy Osharode, Johnson Oji Otutu, Kehinde Johnathan Awatefe, Avwoghokoghene Christopher Idibie, Violet Oshenyen Ejomafuvwe. Multi-instrumental analysis of bioactive compounds and antioxidant properties in Rauvolfia vomitoria Afzel. leaves extract. Int. J. Sec. Metabolite. 2026 Sep. 1;13(3):844-60. doi:10.21448/ijsm.1795990