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Abstract

Newbouldia leavis is a potent herbal plant which holds a deep place in traditional medicine. Its strength is not only in leaf and roots but the chemistry of its stem bark. The stem bark extract contains several phyto-compounds known to possess anti-inflammatory, antitumor, cytotoxic, gastro-protective, anticancer, dermato-protective, anti-helminthic, antioxidant, antidiabetic, cardio-protective, immune-stimulatory, hepato-protective amongst others. GC-MS analysis of bioactive compounds in ethanol stem bark extracts of Newbouldia leavis revealed a total of 36 phyto-compounds representing 96.57 %. The stem bark was dominated by Humulene (16.85 %), gamma-Terpinene (16.11 %), 9, 17-Octadecadienal (12.53 %), p-Cymene (10.59 %), Linoleic acid ethyl ester (9.37 %), 4-Amino-3-hydroxybutanoic acid (8.85 %) and Caryophyllene (7.51 %). A synergy between these compounds heals, nourish and improves the overall vitality of the body.

Introduction

Newbouldia leavis is an evergreen multipurpose herbal plant belonging to the family Bignoniaceae widely distributed in Africa and some parts of Asia [1]. The plant contains unique profile of phyto-compounds with therapeutic properties. In folk medicine, it is used for treating dysentery, pyrexia, gastrointestinal disorder, skin infections, cough, arthritis, tooth ache, bleeding and sexually transmitted diseases [2]. The stem bark and leaves of Newbouldia leavis contains unique profile of flavonoids and phenolic compounds, nature’s compound known to trigger oxidative stress and disrupts cancer cells [3], neutralize free radicals and protects cell membrane from oxidative damage [4]. The stem bark also contains tannins and alkaloids with therapeutic potential that calms, cleanse and restores the body [5].Newbouldia leavis leaves are rich in minerals like calcium, magnesium and phosphorus capable of supporting bone strength and electrolyte balance [6].
Ethanolic extract from the stem bark of Newbouldia leavis have been reported to inhibit Staphylococcusspp, Pseudomonas aeruginosa, Salmonella typhi,Salmonella spp  and Escherichia coli which are the common bacteria behind wound and intestinal infections [7]. Root infusions are used to treat gastrointestinal discomfort, respiratory issues and sexually transmitted diseases [8]. In preclinical studies, extract from Newbouldia leavis leaves, root and stem bark have shown antitumor potential and reduce cancer cell proliferation [9]. Methanolic extract from Newbouldia leavis leaves helps to reduce insulin response and glucose metabolism [10]. Bark decoction are used to treat acute diarrhea, bleeding and inflammed mucus membrane [11-12].
Previous research by [13] have shown that Newbouldia leavis methanolic leaf extract contained 9,12-Octadecadienoic acid (Linoleic acid) (15.35 %), Oleic acid (14.68 %), Stigmasterol(11.12%), 13-Octadecenal (10.98 %), Phytol (6.90 %) and n-Hexadecanoic acid (5.17 %) as predominant phyto compounds. Chemical constituents present in methanol extract of Newbouldia leavis stem bark revealed the presence of Maltol (9.64 %),  2-(3-Bromo-4-methoxy-4 methyl cyclohexyl) propionitrile (8.46 %), 4-((1E)-3-hydroxy-1-propenyl)-2-methoxyphenol (2.25 %) and Benzoicacid,4-hydroxy-3,5-dimethoxy- (2.06 %) as major bioactive compounds [14]. However, there have been inconsistencies in result due to variation in plant age, location, processing method, storage and specie [11]. Therefore, this study was carried out to examine the bioactive compounds in ethanol stem bark extracts of Newbouldia leavis by GC-MS analysis. 

Materials and Methods

Experimental location

The experiment was conducted at the Biochemistry Department, Gandhi College of Agriculture, Rajasthan, India.
 
Collection of Newbouldia leavis stem bark and extraction

Fresh and mature stem bark of Newbouldia leavis was collected from Gandhi Teaching and Research Farm Rajasthan, India in the month of December, 2023 and sent to the Biological Science department, Sumitra Research Institute, Gujarat for proper identification before it was assigned a voucher number SG/08C/2023H. The samples were cut into smaller pieces, air dried for 12 days and powdered using an electric blender. 250 g of Newbouldia leavis stem bark powder was macerated into 1000 mL ethanol for 48 hours at room temperature with intermittent stirring. It was then filtered with Whatman filter paper and concentrated under rotary evaporator at a temperature of 45°C before it was transferred immediately to the laboratory for further analysis. 
 
Gas chromatography – mass spectrometry analysis of Newbouldia leavis stem bark extract

GC/MS analysis of Newbouldia leavis stem bark extract was carried out using Claudus 5006 GC-MS Auto Sampler (China) equipped with two silica capillary columns, interfaced with a quadrupole detector (single quadrupole acquisition Method-MS parameters report), source temperature 230°C, Quadrupole temperature 150°C; the temperature program was 60°C for 2 min,60-240°C at 3 °C/min,then kept at 240 °Cduring 8 min; injector temperature,240°C. The mass spectrometry transfer line temperature, 250 °C; carrier gas, helium at a flow rate of 0.7 ml/min; injection type, split, 20:1; ionization voltage, 70 eV; electron multiplier 1000 eV; scan range 33-400 amu; scan rate, 1.56 scan/s.
Identification of components Interpretation on mass spectrum GC-MS was conducted using the database of National Institute Standard and Technology [15] having more than 62,000 patterns. The spectrum of the unknown component was compared with the spectrum of the known components stored in the NIST library. 

Table 1: Phyto-compounds in Newbouldia leavis stem bark extract by GC-MS analysis

Result and discussion

GC-MS analysis of Newbouldia leavis stem bark extract revealed a total of 36 phyto-compounds representing 96.57 %. The stem bark was dominated by Humulene (16.85%), gamma-Terpinene(16.11%),9,17-Octadecadienal(12.53%), p-Cymene (10.59 %), Linoleic acid ethyl ester (9.37 %), 4-Amino-3-hydroxybutanoic acid (8.85 %) and Caryophyllene (7.51 %) which has been previously associated with antimicrobial, anti-inflammatory, cardio-protective and antioxidant properties [16]. Report by Ezealisiji and Nwoke [17] have shown high concentration of 9,17-Octadecadienoic acid (15.35 %), Oleic acid (14.68 %), n-Hexadecanoic acid (5.17 %) compared to 12.53 %, 0.83 % and 0.02 % recorded in this study for Newbouldia leavis leaf extract. Similarly, Bright et al. [18] recorded a higher concentration of (6.35 %) for n-Hexadecanoic acid and 6.44 % (9,17- Octadecadienal acid) in methanolic extract of Newbouldia leavis stem bark. However, the result on 1-Hexadecanol (0.63 %), 2, 3-Dibromopentane (1.92 %), Cyclopentaneundecanoic acid (0.11 %) and Humulene (16.85 %) observed in this study was similar to those reported by [19-21]. Geographical location, processing method and age of plant have been reported to influence the chemical composition of herbal plants [20-21]. The synergy between the major and minor phyto-compounds exerts numerous therapeutic properties, for instance, Oleic acid, 9-Eicosenoic acid, Linoleic acid ethyl ester and Palmitoleic acid have been suggested to possess antimicrobial properties [22-23], and they also protect the liver, kidney tissues from oxidative stress and reduce damage from toxins [24, 25]. Humulene have been identified to improve cardiovascular health and impede the growth of cancer cells [26-27-30]. 2, 4-Di-tert-butylphenol have been found to possess hypercholesterolemic and protects against neuro-degenerative diseases [28-29-31].

Conclusion

It was concluded that stem bark extracts of Newbouldia leavis contains numerous bioactive compounds with therapeutic properties. These compounds offers incredible healing power by its ability to fight, relieve pain and help to balance immune activities.

References

1. Tanko, Y., M. A. Okasha, M. I. A. Saleh, A. Mohammed, M. Yerima, A. H. Yaro, and A. I. Isa. "Anti-diabetic effect of ethanolic flower extracts of newbouldia laevis (bignoniaceae) on blood glucose levels of streptozocin-induced diabetic Wistar rats." Res J Med Sci 2, no. 2 (2008): 62-65.
   View at Publisher | View at Google Scholar
2. Ogunlana, O. E., and O. O. Ogunlana. "In vitro assessment of antioxidant activity of Newbouldia laevis." Journal of Medicinal Plants Research 2, no. 8 (2008): 176-179.
   View at Publisher | View at Google Scholar
3. Okere, Maureen Chijioke, Onu Uchenna Linda, and Rosemary Uchegbu. "GC-MS, Phytochemical and Antimicrobial Analysis of the Leaf of Newbouldia Laevis (P. Beav)." International Journal of Innovative Research and Development 7, no. 7 (2018).
   View at Publisher | View at Google Scholar
4. Akunyili, D. N. (2000). Anticonvulsant activities of the ethanolic extract of Newbouldia laevis. Paper presented at the 2nd NAAP Scientific conference. 155-158
   View at Publisher | View at Google Scholar
5. Okeke, Amaechina Okechukwu. "Three-minute herbal treatment to reduce dental caries with a Newbouldia laevis based extract." American Journal of Undergraduate Research 2, no. 2 (2003): 1-4.
   View at Publisher | View at Google Scholar
6. De NB, Nanja NA. Studies on Antibacterial Effect of Newbouldia laevis and Aspilia Africana Bagale Journal of Pure and Applied Sciences. 2001; 1(1):24-29.
   View at Publisher | View at Google Scholar
7. Gormann, Rainer, Maki Kaloga, Daneel Ferreira, Jannie PJ Marais, and Herbert Kolodziej. "Newbouldiosides A–C, phenylethanoid glycosides from the stem bark of Newbouldia laevis." Phytochemistry 67, no. 8 (2006): 805-811.
   View at Publisher | View at Google Scholar
8. Chen, Yisheng, Caihong Huang, Bernhard Hellmann, Zhengyu Jin, Xueming Xu, and Guosheng Xiao. "A new HPTLC platformed luminescent biosensor system for facile screening of captan residue in fruits." Food Chemistry 309 (2020): 125691.
   View at Publisher | View at Google Scholar
9. Ali, Saima, Muhammad Rashid Khan, Irfanullah, Moniba Sajid, and Zartash Zahra. "Phytochemical investigation and antimicrobial appraisal of Parrotiopsis jacquemontiana (Decne) Rehder." BMC complementary and alternative medicine 18, no. 1 (2018): 43.
   View at Publisher | View at Google Scholar
10. Wagh, Anita S., and SANTOSH R. Butle. "Plant profile, phytochemistry and pharmacology of Spathodea campanulata P. Beauvais (African tulip tree): a review." International Journal of Pharmacy and Pharmaceutical Sciences 10, no. 5 (2018): 1-6.
    View at Publisher | View at Google Scholar
11. Kushwaha, S and A.Maurya (2018). “Natural products: an important source for the treatment of psychosis,” Archives of Organic and Inorganic Chemical Sciences, vol. 3, no. 3, pp. 353–355, 2018
    View at Publisher | View at Google Scholar
12. Simoben, Conrad V., Fidele Ntie-Kang, Sergi H. Akone, and Wolfgang Sippl. "Compounds from African medicinal plants with activities against selected parasitic diseases: schistosomiasis, trypanosomiasis and leishmaniasis." Natural products and bioprospecting 8, no. 3 (2018): 151-169.
    View at Publisher | View at Google Scholar
13. Utzinger, Jürg, Jennifer Keiser, Xiao Shuhua, Marcel Tanner, and Burton H. Singer. "Combination chemotherapy of schistosomiasis in laboratory studies and clinical trials." Antimicrobial agents and chemotherapy 47, no. 5 (2003): 1487-1495.
    View at Publisher | View at Google Scholar
14. Loganathan, V., M. Devi Kaniakumari, and P. Selvakumar (2017). A study of the physico-chemical and phytochemical parameters of leaves of Mallotus rhamnifolius,” International Journal of Pharmacognosy and Phytochemical Research, vol. 9, no. 6, pp. 858–863, 2017.
    View at Publisher | View at Google Scholar
15. NIST. Chemistry Web Book. Data from NIST Standard Reference Database 69, 2011.
    View at Publisher | View at Google Scholar
16. Anaduaka, Emeka G., Victor N. Ogugua, Simeon I. Egba, and Victor O. Apeh. "Investigation of some important phytochemical, nutritional properties and toxicological potentials of ethanol extracts of Newbouldia laevis leaf and stem." African Journal of Biotechnology 12, no. 40 (2013).
    View at Publisher | View at Google Scholar
17. Maduabuchi, Ezealisiji Kenneth, and Nwoke Emmanuel Awucha. "Phytochemistry and GC-MS analysis of methanolic leaf extract of Newbouldia leavis (Bignonacea)." Journal of Pharmacognosy and Phytochemistry 9, no. 2 (2020): 1998-2004.
    View at Publisher | View at Google Scholar
18. Osei-Mensah, Bright, Yaw Duah Boakye, William Kofi Anyan, Theresa Appiah Agana, Eugene Agyei Aboagye, Ivy Bentil, Elvis Suatey Lomotey, Francis Adu, and Christian Agyare. "In Vitro Cercaricidal Activity, Acute Toxicity, and GC/MS Analysis of Some Selected Ghanaian Medicinal Plants." Journal of Parasitology Research 2023, no. 1 (2023): 4589424.
    View at Publisher | View at Google Scholar
19. Okere, Maureen Chijioke, Onu Uchenna Linda, and Rosemary Uchegbu. "GC-MS, Phytochemical and Antimicrobial Analysis of the Leaf of Newbouldia Laevis (P. Beav)." International Journal of Innovative Research and Development 7, no. 7 (2018).
    View at Publisher | View at Google Scholar
20. "Phenolic content and DPPH radical scavenging activity of yam-containing surimi gels influenced by salt and heating." Asian Journal of Health and Information Sciences 2, no. 1-4 (2007): 1-11.
    View at Publisher | View at Google Scholar
21. Lima, A. L., R. A. M. E. N. D. U. Parial, M. A. N. I. S. H. A. Das, and ASISH KUMAR Das. "Phytochemical and pharmacological studies of ethanolic extract from the leaf of mangrove plant Phoenix paludosa Roxb." Malaysian Journal of Pharmaceutical Sciences 8, no. 2 (2010): 59-69.
    View at Publisher | View at Google Scholar
22. Ajayi, I. A., O. Ajibade, and R. A. Oderinde. "Preliminary phytochemical analysis of some plant seeds." Research Journal of Chemical Sciences 1, no. 3 (2011): 58-62.
    View at Publisher | View at Google Scholar
23. Shittu, M. D., and J. O. Alagbe. "Phyto-nutritional profiles of broom weed (Sida acuta) leaf extract." Annals of Clinical Toxicology 3, no. 2 (2020).
    View at Publisher | View at Google Scholar
24. Alagbe, J. O., E. U. Anaso, D. N. Anorue, and H. Zubairu. "Influence of Chromolaena Odorata Extract on the Growth Performance and Fatty Acid Composition of Weaner Rabbits." International Journal of Advanced Technology and Social Sciences 3, no. 8 (2025): 1199-1212.
    View at Publisher | View at Google Scholar
25. Sharma, Singh, Alagbe Olujimi John, Liu Xing, Sharma Ram, and Kumar Amita. "Comparative analysis of ethanolic Juniperus thurifera leaf, stem bark and root extract using gas chromatography and mass spectroemetry." International Journal of Agriculture and Animal Production 2, no. 6 (2022): 18-27.
    View at Publisher | View at Google Scholar
26. Adewale AO, Alagbe JO, Adeoye, AO. Dietary Supplementation of Rauvolfia Vomitoria Root Extract as A Phytogenic Feed Additive in Growing Rabbit Diets: Haematology and serum biochemical indices. International Journal of Orange Technologies, 2021; 3(3): 1-12.
    View at Publisher | View at Google Scholar
27. Alagbe, J. O. "Gas chromatography and mass spectroscopy of Juniperus phoenice stem bark extract and its influence on the haemato-biochemical values of growing rabbits." British Scientific Periodical 1, no. 1 (2022): 18-33.
    View at Publisher | View at Google Scholar
28. Choi, Jae-Suk, Nam-Hee Park, Seon-Yeong Hwang, Jae Hak Sohn, Inseok Kwak, Kwang Keun Cho, and In Soon Choi. "The antibacterial activity of various saturated and unsaturated fatty acids against several oral pathogens." Journal of Environmental Biology 34, no. 4 (2013): 673.
    View at Publisher | View at Google Scholar
29. Zheng, Chang Ji, Jung-Sung Yoo, Tae-Gyu Lee, Hee-Young Cho, Young-Ho Kim, and Won-Gon Kim. "Fatty acid synthesis is a target for antibacterial activity of unsaturated fatty acids." FEBS letters 579, no. 23 (2005): 5157-5162.
    View at Publisher | View at Google Scholar
30. Morris, Hernández, and Alagbe John Olujimi. "INFLUENCE OF Odontonema Strictum OIL ON THE GROWTH PERFORMANCE AND RUMINAL FERMENTATION OF BARBARI BUCKS." Research in: Agricultural & Veterinary Sciences 9, no. 2 (2025).
    View at Publisher | View at Google Scholar
31. Alagbe, John Olujimi. "Impact of dietary supplementation of Rhamnus prinoides leaf extract on the growth performance, nutrient retention and intestinal microbial count of “japanese quails”." Brazilian Journal of Science 3, no. 5 (2024): 40-50.
    View at Publisher | View at Google Scholar