Plants are the basis of traditional medicine in Africa and have been used for thousands of years. These plants often exhibit a wide range of pharmacological activities [1]. Due to the need for development of new drugs with better pharmacological activities, dependence on plants grew increasingly as scientists continuously exploited them for isolation of bioactive compound.

Spondias mombin is a small tree that grows up to 20 m (60 ft.) high and 1.5 m (5 ft) in girth, moderately buttressed; bark thick, corky, deeply fissured, slash pale pink, darkening rapidly, branches low, branchlets glabrous; leaves pinnate, leaflets 5-8 opposite pairs with a terminal leaflet. It belongs to the family Anacardiaceae. It flowers between January to May and fruits between July to September. The fruits have a sharp, somewhat acid taste and are edible. The matured fruit has a leathery skin and a thin layer of pulp. The fruit pulp is either eaten fresh, or made into juice, concentrate, jellies, and sherbets. The fruit-juice is used as a febrifuge and diuretic. The roots are also used as febrifuge in Ivory Coast. The stem bark is used as a purgative and in local applications for leprosy. The stem bark decoction is also used in the treatment of severe cough. It serves as an emetic, a remedy for diarrhea, dysentery, haemorrhoids and a treatment for gonorrhoea and leucorrhea [2].

A report showed that the bark contains a certain amount of tannin and this explains the reason why the dry pulverized bark is applied as a dressing to a wound [2]. In Belize, a decoction of the young leaves is a remedy for diarrhea and dysentery. The juice of crushed leaves and the powder of dried leaves are used as poultices on wounds and inflammations. The gum is employed as an expectorant and to expel tapeworms [3].

A decoction of the mashed leaves is used by the Ibos (Nigeria) for washing a swollen face. A leaf infusion is a common cough remedy or used as a laxative for fever with constipation. A leaf decoction is used in treatment of gonorrhea. The leaves are used in Ivory Coast for fresh wounds to prevent inflammation. A decoction of pounded leaves of S. mombin is used as an eye lotion and the juice pressed from young, warm leaves is given to children for stomach troubles. The extract has shown anti-inflammatory activity in Wistar rats [4]. A tea made from the flowers and leaves is taken to relieve stomach ache, biliousness, and urethritis, cystitis and eye and throat inflammations. A decoction of the root is used as purgative [4].

Collection of plant materials

The stem bark of Spondias mombin (Linn.) was collected from a local farm at Owo (710’59.998N and 534’59.988E), Ondo State, Nigeria at WAT UTC+1 time zone on 26^th and 27^th of February, 2016. Fresh and healthy plants were also collected during its fruiting season, between April and July 2017, from the same geographical locations.

Authentication of Spondias mombin (Linn.)

The plants were authenticated at the Department of Plant Science and Biotechnology, Adekunle Ajasin University, Akungba- Akoko, Ondo state, Nigeria.

Preparation and extraction of Spondias mombin (Linn.) Plant

The stem-bark of Spondias mombin plant were harvested and airdried. The dried stem bark were milled into powdered form using manual grinder. Powdered plant material (1 kg) each of the different plant parts was extracted with 3 L of 70% (v/v) ethanol, ethyl acetate and distilled water for 72 h at room temperature. The extraction process was repeated four times until the extract became clear. The filtrates were combined and concentrated under reduced pressure rotatory evaporator at 35°C to give, SMSBEA for the stem-bark part. The dry extracts were kept in tightly stoppered bottles in a refrigerator at -4°C for further analysis

The ethyl acetate fraction SMSBEAEA (11.0 g) was adsorbed unto silica and allowed to dry before packing on to column. The column was wetted with n-hexane and gradient elution effected with the following solvent/ solvent mixtures (Table 1).

Table 1: Fractionation of SMSBEAEA on Column Chromatography with Hexane/ethyl acetate/ Methanol solvent.

Fractions collected were analysed by TLC (Thin layer chromatography) in Hex - EtOAc - AcOH (1: 9: 0.5). The resulting spots on TLC plates 1-3 were visualized under UV light (254 nm) and detected by the use of vanillin/sulfuric acid and the antioxidant compound(s) were detected using DPPH spray reagents. Fractions having the same TLC patterns were bulked, concentrated in vacuo to dryness and weighed; resulting in three fractions coded SMSBEAEA1, SMSBEAEA2 and SMSBEAEA3 (Table 2).

Table 2: Three fractions coded SMSBEAEA1, SMSBEAEA2 and SMSBEAEA3 final Weight after Column Chromatography.

TLC Plate 1: Isolation, Identification and Characterization of Epigallocatechin, and Epicatechin.

TLC Plate 2: Isolation, Identification and Characterization of Stigmasterol Phytosterol.

TLC Plate 3: Isolation, Identification and Characterization of Stigma sterol Phytosterol.

Fractionation of SMSBEAEA1 on Sephadex LH-20

Fraction SMSBEAEA1 (1.6 g) was dissolved in a small amount of CHCl₃-MeOH (70:30) solvent mixture and loaded on a Sephadex LH-20 column previously equilibrated with the same solvent mixture and elution was isocratically effected. Fractions (about 20 ml each) collected were analysed by TLC in CH₂Cl₂ - MeOH (10: 1.0) and fractions having the similar TLC patterns were bulked together, concentrated in vacuo to dryness and weighed; resulting in eight fractions coded SMSBEAEA1a, SMSBEAEA1b, SMSBEAEA1c (compound 1), SMSBEAEA1d, SMSBEAEA1e (compound 2), SMSBEAEA1f, SMSBEAEA1g and SMSBEAEA1h (Table 3).

Table 3: Eight fractions coded SMSBEAEA1a, SMSBEAEA1b, SMSBEAEA1c (compound 1), SMSBEAEA1d, SMSBEAEA1e (compound 2), SMSBEAEA1f, SMSBEAEA1g and SMSBEAEA1h final Weight on Sephadex LH-20.

Fractionation of SMSBEAEA1a on column Chromatography

Fraction SMSBEAEA1a (600 g) was adsorbed unto silica and allowed to dry before packing on to column. The column was wetted with n-hexane and gradient elution effected with the following solvent/solvent mixtures (Table 4):

Table 4: Fractionation of SMSBEAEA on Column Chromatography with Hexane/dichloromethane solvent

Fractions (about 20 ml each) collected were analysed by TLC in hexane - CH₂Cl₂ (8:4) and hexane - CH₂Cl₂ (2:8) and fractions having the similar TLC patterns were bulked together, concentrated in vacuo to dryness and weighed resulting in six fractions coded SMSBEAEA1a1, SMSBEAEA1a2, SMSBEAEA1a3, SMSBEAEA1a4 (3), SMSBEAEA1a5 and SMSBEAEA1a6 (Table 5 and Figure 1).

Table 5: Six fractions coded SMSBEAEA1a1, SMSBEAEA1a2, SMSBEAEA1a3, SMSBEAEA1a4 (3), SMSBEAEA1a5 and SMSBEAEA1a6 final Weight on Sephadex LH-20

Figure 1: Isolation and purification of active compounds from the bioactive fractions (Pure compound) Key: 1- Epigallocatechin; 2- Epicatechin and 3-Stigmasterol Phytosterol.

The physical and spectroscopic data on the isolated compounds are as recorded below.

Characterization of compound 1

Physical characteristics: Pale yellow powder, m.p. 240-242°C (decomp); [α]_D²⁰ -60 (CD₃OD, c=1.00); ESI-MS m/z 291 [M+H]⁺, 139 [M+H-152]⁺; IR Vmax KBr cm^-1: 3456.2, 1620.10, 1521.70, 1450.40, 1265.2 1143.7; UV λmax MeOH nm 213.00, 280; ¹H- and ¹³C-NMR (CD₃OD).

Spectroscopic data 1: ¹Hnmr (300 MHz, CD₃OD) δ ppm: 4.89 (1H, bs, H-2), 4.01 (1H, bs, H-3), 2.88 (1H_a, dd, J=4.1, 12.15 Hz, H-4a), 2.84, 1H_b, dd, (4.5, 12.09 Hz, H-4b), 5.89 (1H, d, J=2.2 Hz, H-6), 5.97 (1H, d, J=2.2 Hz, H-8), 7.09 (1H, d, J= 2.2 Hz, H- 2′), 6.74 (1H, d, J= 6.00 Hz, H- 5′), 6.79 (1H, dd, J= 6.2, 1.9 Hz, H-6′).

Spectroscopic data 2: ¹³Cnmr (75 MHz, CD₃OD) δ ppm: 82.8 (C- 2), 68.8 (C-3), 28.5 (C-4), 157.5 (C-5), 95.6 (C-6), 157.7 (C-7), 96.5 (C- 8), 156.8 (C-9), 101.0 (C-10), 132.2 (C-1′), 115.4 (C-2′), 146.2 (C-3′), 146.4 (C-4′), 116.3 (C-5′), 120.2 (C-6′).

The ¹Hnmr, ¹³Cnmr, DEPT, HMQC, HMBC, MS, IR and UV spectra are provided in Figure 1.

Characterization of compound 2

Physical characteristics: Brown amorphous powder, [D²⁰ 110⁰ (Me₂CO, c=1.00); EIMS m/z ESI-MS m/z 345 [M+K]⁺, 139 [M+H-168]⁺; IR V_max KBr cm^-1: 3345, 2923.45.9, 1614.3, 1519.8, 1463.9, 1353.9, 1282.6, 1195.8; ¹H- and ¹³C-NMR (CD3OD).

Spectroscopic data 1: ¹Hnmr (300 MHz, CD₃OD) δ ppm: 4.89 (1H, bs, H-2), 4.01 (1H, bs, H-3), 2.88 (1H_a, dd, J=4.1, 12.15 Hz, H-4a), 2.84, 1H_b, dd, (4.5, 12.09 Hz, H-4b), 5.89 (1H, d, J=2.2 Hz, H-6), 5.97 (1H, d, J=2.2 Hz, H-8), 6.9 (2H, s, H- 2′ / H-6′).

Spectroscopic data 2: ¹³Cnmr (75 MHz, CD₃OD) δ ppm: 82.8 (C- 2), 68.8 (C-3), 28.5 (C-4), 157.5 (C-5), 95.6 (C-6), 157.7 (C-7), 96.5 (C- 8), 156.8 (C-9), 101.0 (C-10), 132.2 (C-1′), 110.5 (C-2′), 146.2 (C-3′), 132.6 (C-4′), 146.4 (C-5′), 110.5 (C-6′).

The ¹Hnmr, ¹³Cnmr, COSY, DEPT, HMQC, HMBC, MS, IR and UV spectra are provided as Figure 2.

Figure 2: MeOD of Epicatechin.

Characterization of compound 3

Physical characteristics: White crystal, ESI-MS m/z: [M + H] 413, ¹H- and ¹³C-NMR (CDCl₃)

Spectroscopic data 1: ¹³Cnmr (75 MHz, CDCl₃) d ppm: 30.79 (C- 1), 30.18 (C-2), 69.26 (C-3), 38.42 (C-4), 146.29 (C-5), 117.74 (C-6), 37.56 (C-7), 41.12 (C-8), 48.78 (C-9), 31.66 (C-10), 21.65 (C-11), 39.34 (C-12), 42.63 (C-13), 56.51 (C-14), 24.08 (C-15), 25.00 (C-16), 54.52 (C-17), 12.87 (C-18), 14.56 (C-19), 40. 80 (C-20), 21.49 (C-21), 132.46 (C-22), 129.94 (C-23), 51.35 (C-24), 30.99 (C-25), 21.29 (C-26), 19.14 (C-27), 25.23 (C-28), 11.57 (C29) (Figures 3-5).

Figure 3: APT of Epicatechin.

Figure 4: MeOD of Epigallocatechin.

Figure 5: APT of Epigallocatechin.

Spectroscopic data 1: The ¹³Cnmr, MS, spectra (fig 2). ¹³Cnmr (75 MHz, CD₃OD) δ ppm: 82.8 (C-2), 68.8 (C-3), 28.5 (C-4), 157.5 (C-5), 95.6 (C-6), 157.7 (C-7), 96.5 (C-8), 156.8 (C-9), 101.0 (C-10), 132.2 (C- 1′), 110.5 (C-2′), 146.2 (C-3′), 132.6 (C-4′), 146.4 (C-5′), 110.5 (C-6′).

This research work is based on the isolation, purification and elucidation of structure (Chemical characterization) of the isolated constituents from Spondias mombin extract. It was observed that, three different bioactive constituents were isolated, purified and structurally elucidated from the stem bark extract of Spondias mombin. They are Epigallocatechin, Epicatechin and Stigmasterol Phyto sterol.

Identification of Compound 1 as 3, 5, 7, 3', 4'-pentahydroxyflavanol (Epicatechin)

From the ethyl acetate fraction of the stem bark, compound 1 was isolated as yellow solid. The negative ESI –MS spectrum of compound 1 showed ion peak at m/z 291 [M + H]⁺ corresponding to the molecular formula C₁₅H₁₄O₆. In the same MS spectrum, fragment ion found at m/z 139 resulted from a retro-diels-Alder (RDA) fragmentation. Analyzing the ¹³C NMR and ¹H NMR spectral data revealed presence of two aromatic systems one exhibiting an ABD proton spin system with protons resonating at δ_H 7.09 (1H, d, J = 1.8 Hz), δ_H 6.74 (1H, d, J = 6.0 Hz) and 6.79 (1H, dd, J = 6.2, 1.9 Hz) assignable to H-2', H-5' and H-6' protons attached on carbons C-2' (δ_C 115.4), C-5' (δ_C 116.3) and C-6' (δ_C 120.2) on ring B respectively. The HMBC spectrum showed that H-2' proton correlated with carbons C-2 (δ_C 78.9), C-3' (δ_C 146.2), C-4' (δ_C 146.4), C-5' and C-6' whereas H-6' proton showed correlations with carbons C-1' (δ_C 132.2), C-2, C- 3', C- 4', C-5' and C-6' (Figures 6-13).

Figure 6: COSY analysis of compounds isolated from Stem bark extract of Spondias mombin.

Figure 7: HMBC analysis of compounds isolated from Stem bark extract of Spondias mombin.

Figure 8: HSQC analysis of compounds isolated from Stem bark extract of Spondias mombin.

Figure 9: Noesy analysis of compounds isolated from Stem bark extract of Spondias mombin.

Figure 10: MS analysis of compound A1 isolated from Stem bark extract of Spondias mombin (SpectrumR.Time:1.067(Scan#:65) MassPeaks:2005).

Figure 11: FTIR of Epicatechin.

Figure 12: FTIR of Epigallocatechin.

Figure 13: FTIR of Stigmasterol Phytosterol.

Furthermore the HMBC spectrum showed correlations of H-5' proton with carbons C-1', C-3' and C-4'. Thus the ABD proton spin system was placed on ring B. Ring C on the other hand displayed a saturated system with two germinal protons resonating at δ_H 2.88 (1H_a, dd, J = 4.1, 12.15 Hz) and 2.84 (1H_b, dd, J = 4.5, 12.09 Hz) and they were placed at C-4 (δ_C 28.5) based on DEPT, HMQC and HMBC experiments. The negative ESI –MS fragment ion at m/z 139 resulting from a retro Diels – Alder fragmentation (Scheme 1) confirmed that carbon C-4 consists of two protons consistent with the flavan skeleton. The absence of carbonyl absorption peaks in the ¹³C NMR spectrum (Figures 6-9) on the region 170 – 210 ppm (Agrawal,1989) and IR around 1700 – 1750 cm-1 confirmed the proposed flavanol skeleton. The second aromatic ring exhibited a proton spin system comprised of meta-coupled protons with resonances observed at δ_H 5.89 (1H, d, J = 2.2 Hz) and 5.97 (1H, d, J = 2.2 Hz) which were assigned to H-6 and H-8 protons placed on carbons C-6 (C 95.6) and C-8 (δ_C 96.5) respectively based on HMBC, DEPT and HMQC experiments, this proton spin system was thus placed on ring A.

Scheme 1: Fragmentation pathways for compound 1 (RDA = retro-Diels-Alder).

From the above information compound 1 was identified as 3, 5, 7, 3', 4'–pentahydroxyflavanol, commonly known as (-) – epicatechin, a widely distributed compound in the plant kingdom and has been reported to be responsible for anti-inflammatory and antioxidant properties in green teas [5] and other plant extracts. However this is the first time we report its occurrence from Spondias mombin (Scheme 1).

¹H and ¹³C NMR Data of Compound 1 were listed as follows - Position 2 (δ_H- 4.89, 1H, br s)( δ_C-82.8), Position 3(δ_H-4.01, 1H, br s)( δ_C-68.8), Position 4((δ_H- 2.88, 1H_a, dd, (4.1, 12.15 Hz, 2.84, 1H_b, dd, (4.5, 12.09 Hz)( δ_C-28.5), Position 5 ( δ_C-157.5), Position 6 (δ_H-5.89, 1H, d, (2.2 Hz)( δ_C-95.6), Position 7( δ_C-157.7), Position 8 (δ_H-5.97, 1H, d, (2.2 Hz))( C-96.5), Position 9 (δ_H-)( δ_C- -156.8 ), Position 10 (H-)( δ_C-101.0), Position 1' (δ_H-)( δ_C-132.2), Position 2' (δ_H-7.09, 1H, d, (2.2 Hz)( δ_C-115.4), Position 3' (δ_H-) ( δ_C-146.2), Position 4' (δ_H-)( δ_C-146.4), Position 5' (δ_H-6.74, 1H, d, (6.0 Hz)( δ_C-116.3), Position 6' (δ_H-6.79, 1H, dd, (6.2, 1.9 Hz)) ( δ_C-120.2), Assignments were confirmed by HMBC, HMQC, DEPT and COSY experiments

Identification of Compound 2 as Epigallocatechin

This compound was obtained as orange powder and gave a molecular ion peak at m/z at 306.07 in EIMS which is compatible with the molecular formula C₁₅H₁₄O₇. Another fragment ion was found at m/z 139.04 which resulted from a retro-Diels–Alder (RDA) fragmentation.

The ¹H-NMR spectrum of 2 showed signals due to the presence of a pyrogallol ring at 6.53, 2H, s) which was assigned to protons H-2′ /H-6′ placed on carbons C-2′ / C-6′ (δ_C 105.59) in ring B, a phloroglucinol ring comprising of two meta - coupled protons observed at δ_H 5.96 (1H, d, J = 2.4 Hz) and 5.93 (1H, d, J = 2.4 Hz) which were assigned to protons H-6 and H-8 placed on carbons C-6 (δ_C 94.98) and C-8 (C 94.50) respectively, two non –aromatic oxygen bearing methane 4.76, and 4.18 which were assigned to protons H-2 and H-3, a methylene signal at 2.86 and 2.74 which was assigned to H-4. By the combination of ¹H-NMR, ¹³C-NMR, DEPT, HMQC and HMBC spectral analysis, all carbon and proton signals could be definitely assigned.

Based on the above evidence, compound 2 were concluded to be epigallocatechin which was in agreement with the reported literature values [6]. It is a broadly distributed chemical constituent in the plant kingdom and is reported to possess a powerful antioxidant activity [7].

Compound 2

¹H- and ¹³C-NMR Spectral Data for compound 2 in CD₃OD at 300 MHz were listed below.

Ring C- C-2 (δ_H-4.89, 1H, br s )( δ_C-82.8), ¹³C NMR(in acetone-d₆)- 79.4, C-3 (δ_H-4.01, 1H, br s)) δ_C-68.8), ¹³C NMR(in acetone-d₆)- 67.0, C-4 (δ_H-2.88, 1Ha, dd, (4.1, 12.15 Hz, 2.84, 1Hb, dd, (4.5, 12.09 Hz)( δ_C-28.5), ¹³C NMR(in acetone-d₆)- 28.8, Ring A- C-5 (δ_H- )( δ_C-157.5), ¹³C NMR(in acetone-d₆)- 157.1, C-6 (δ_H-5.89, 1H, d, (2.2 Hz)) δ_C-95.6, ¹³C NMR(in acetone-d₆)- 96.3, C-7 (δ_H-)( δ_C-157.7), ¹³C NMR(in acetone-d₆)- 157.5, C-8 (δ_H-5.97, 1H, d, (2.2 Hz))( δ_C-96.5), ¹³C NMR(in acetone-d₆)- 95.7, C-9 (δ_H-)( δ_C-156.8), ¹³C NMR(in acetone-d₆)- 157.5, C-10(δ_H-)( δ_C- 101.0), ¹³C NMR(in acetone-d₆)- 99.9, Ring B- C-1′ (δ_H- )( δ_C-132.2), ¹³C NMR(in acetone-d₆)- 131.5, C-2′ (δ_H- 6.9, 1H, s)( δ_C-110.5), ¹³C NMR(in acetone-d₆)- 107.0, C-3′ (δ_H- )( δ_C-146.2), ¹³C NMR(in acetone-d₆)- 146.1, C-4′ (δ_H- )( δ_C-132.60), ¹³C NMR(in acetone-d₆)- 132.9, C-5′ (δ_H- )( δ_C- 146.4), ¹³C NMR(in acetone-d₆)- 146.1, C-6′ (H-6.9, 1H, s)( δ_C-110.5), ¹³C NMR(in acetone-d₆)- 107.0, (Scheme 2).

Scheme 2: ESI–MS fragmentation pattern for compound XII (RDA = retro-Diels-Alder fission (–)-epigallocatechin).

Identification of compound 3 as Stigmasterol phytosterol. Compound 3 was obtained as a white crystal. The positive- ion ESI-MS exhibited [M + H]⁺ at m/z 413 compatible with the molecular formula C₂₉H₄₈O. Its ¹H-nmr and ¹³C-nmr spectrum showed three olefinic protons as one triplet at 5.23 (C-6, 117.74), and two doublets of doublets at 5.16 (C-22, 132.46) and 5.10 (C-23, 129.94), a proton germinal with hydroxyl group as a multiplet at 3.60 (C-3, 71.45). The ¹³Cnmr spectra showed 29 carbon signals composed of 6 methyl, 6 methylene, 3 olefinic methines, 8 non-olefinic methines and 3 quaternary carbons. The identity of compound 3 as stigmasterol was confirmed by comparison of its physical and spectra data with those reported in the literature [8-10]. Stigmasterol is a ubiquitous phytosterol, occurring naturally in a wide variety of plants. However, it is here reported isolated from the plant for the first time.

¹H- and ¹³C-NMR Spectral Data for compound 3 in CD₃Cl at 300 MHz were listed below.

Position 1-¹³C (30.18)( ¹³C NMR-in CD₃Cl-31.90), Position 2-¹³C (69.26)( ¹³C NMR-in CD₃Cl-71.45), Position 4-¹³C (38.40)( ¹³C NMRin CD₃Cl-38.42), Position 5-¹³C (146.29)( ¹³C NMR-in CD₃Cl-139.96), Position 7-¹³C (37.66)( ¹³C NMR-in CD₃Cl-37.56), Position 8-¹³C (41.12) ( ¹³C NMR-in CD₃Cl-41.20), Position 9-¹³C (48.78) ( ¹³C NMRin CD₃Cl-49.88), Position 10-¹³C (31.66) ( ¹³C NMR-in CD₃Cl-34.63), Position 11-¹³C (21.65) ( ¹³C NMR-in CD₃Cl-21.96), Position 12-¹³C (39.34) ( ¹³C NMR-in CD₃Cl-39.88), Position 13-¹³C (42.63)( ¹³C NMRin CD₃Cl-43.70), Position 14-¹³C (56.51) ( ¹³C NMR-in CD₃Cl-56.33), Position 15-¹³C (24.08) ( ¹³C NMR-in CD₃Cl-23.41), Position 16-¹³C (25.00) ( ¹³C NMR-in CD₃Cl-28.88), Position 17-¹³C (54.52) (¹³C NMRin CD₃Cl-55.54), Position 18-¹³C (12.87) ( ¹³C NMR-in CD₃Cl-12.62), Position 19-¹³C (14.56) ( ¹³C NMR-in CD₃Cl-13.42), Position 20-¹³C (40.18) ( ¹³C NMR-in CD₃Cl-40.68), Position 21-¹³C (21.49) ( ¹³C NMRin CD₃Cl-21.46), Position 22-¹³C (132.46) ( ¹³C NMR-in CD₃Cl-138.55), Position 23-¹³C (129.94) ( ¹³C NMR-in CD₃Cl-129.88), Position 24-¹³C (51.35) ( ¹³C NMR-in CD₃Cl-51.65), Position 25-¹³C (30.99) ( ¹³C NMRin CD₃Cl-32.27), Position 26-¹³C (21.29) ( ¹³C NMR-in CD₃Cl-21.76), Position 27-¹³C (19.14) ( ¹³C NMR-in CD₃Cl-19.37), Position 28- ¹³C (25.23) ( ¹³C NMR-in CD₃Cl-25.78), Position 29-¹³C (11.57) ( ¹³C NMR-in CD₃Cl-12.44), (Idowu, 2016). All the three pure compounds are well known but reported for the first time in Spondias mombin.

Compound 3

Some authors classified epicatechin as a type of flavonoid which is found in Spondias mombin extract, polyphenols comprises of 30-40% of extract solids in the herbaceous tree, there are different classes of epicatechin, epigallocatechin [11]. Epicatechin have been proven to have diverse benefits to human health. It reduces the risks of diabetes mellitus and cardiovascular diseases this shows the basic important aspect of Spondias mombin tree, which have pharmalogical effects such as anti hyporlipidemic, anti-inflammatory, antioxidative effect, anticarcinogenic and cytoprotective [7].

EGC arts as a strong inhibitor of HIV replication in cultured peripheral blood cells and inhibition of HIV-1 reverse transcriptase in vitro. EGC binds directly to CD4 molecules with consequent inhibition of Gp 120 binding and inactivate viruses in vitro by deformation of phospholipids [12-14]. It should be mentioned that EC are antioxidant, they are effective scavengers and free radical such a reactive oxygen species (ROS), Reactive nitrogen species and superoxide [15].

Epicatechin and other polyphenols decreases the susceptibility of low density lipoprotein to oxidation which prevents the initiation of artherosclerosis, HIV protein (Tat) and gp120 is known to cause neurotoxicity in human via mechanisms that activate macrophages and glial cells and finally, oxidative stress it can be suggested that epicatechin are neuroprotective by blocking the neurotoxic effects of the HIV protein which cause oxidative stress [16].

However, it should be mentioned that Catechins have an antiprohferative effect on turmor cells as well as inhibiting metastasis and it ability to modulate antioxidant enzymes in vivo. It has been shown to increase the activity of super oxide dismutase and catalase. It also suppresses lipid peroxidation of turmor cells [17].

It must be clearly stated that stigmasterol phytosterol have been shown to lower/reduce blood cholesterol and this lowering may reduce the risk of coronary heart disease. Phytosterols are under preliminary research for their potential to inhibit lung, stomach, ovarian and breast cancers as well as colon and prostate cancers [15].

Apart from lipid peroxidation, epicatechin binds secondary bile acid (Tauro-deoxycholic acid), thus secondary bile has been associated with increased risk of developing colorectal cancer thence these epicatechin from Spondias mombin polyphenolic compound may reduce the risk factor for the developing colorectal cancer [18].

In conclusion, epicatechin, epigallocatechin and stigmasterol phytosterol isolated from stem bark of Spondias mombim extract has proven itself to be nature’s extraordinary therapeutic agent and it has also been proved during the course of this project work. It has also been proved that, Spordias mombin is a potent medicinal plant that its uses and application should be encouraged. More time and attention should be spent on developing it as a sustainable drug for prophylaxis and treatment of complications and diseases.

The authors wish to express their appreciation to all the technical staffs of the laboratory unit of Both the Department of Microbiology, Faculty of Science, Adekunle Ajasin University, Akungba Akoko, Ondo State, Nigeria and Obafemi Awolowo University, Ile Ife, Osun State, Nigeria for their support and all the technical assistance rendered during the course of this research work.