Phytochemicals, radical scavenging and metal chelating properties of methanolic extract of Senna alata were determined by various methods to identify the quantity of each phytochemical content present in the plant through standard methods. The phenol content of Senna alata extract was found to be (18.0 ±2.82mgGAE/g) while flavonoid was found to be (10.9± 0.0 mgQAE/g ) showing minimal amounts of flavonoid as compared to phenol. The total antioxidant in Senna alata extract was found to be (13.6 ± 0.08 mgAAE/g). The percentage Copper (Cu2+) Chelation Assay of Senna alataat showed higher percentages at different concentration. The percentage Fe2+ Chelation of Senna alata also showed higher percentage as compared with EDTA. Therefore methanolic extract of Senna alata leaves can then be a very useful source of antioxidant for combating diseases that are linked with free radical and certain metals.
S. alata is found throughout Africa, including Ghana, Brazil, Australia, Egypt, India, Somalia, Sri Lanka, and other countries [1]. The entire plant is used to cure impetigo, ulcers, helminthiasis, and as a purgative [2]. Senna alata leaf extract has been discovered to help reduce blood sugar levels . Senna alata is also used to treat gastrointestinal infections, intestinal worms, typhoid fever, poison, hepatitis, yellow fever [3], wounds, and viral infections [3]
Chemicals
Methanol, Folin-ciocalteu reagent, sodium carbonate, Gallic acid, 1, 10-phenanthroline, iron (II) sulfate (FeSO4), DPPH (1, 1-diphenyl–2 Picrylhydrazyl), sodium nitroprusside, griess reagent, copper sulfate (CuSO4),pyrocatechol, thiourea, acetic acid, ethanol , ammonia solution, Quercetin, sodium nitrite (NaNO2), aluminum chloride (AlCl3), sodium hydroxide (NaOH), ,L-ascorbic acid, sodium phosphate, ammonium molybdate.
PLANT MATERIAL
Fresh leaves of Senna alata were collected and identified by the Department of Botany, Niger Delta University, Bayelsa State. Plant was collected in November, 2021 in the pharmacognosy botanical garden.
METHODS
PREPARATION OF METHANOIC EXTRACT OF SENNA ALATA
Senna alata was harvested and dried in the shade for 14 days (Two weeks). The dried leaves were later grounded to a fine powder. Crushed leaves were soaked in 1500ml methanol for three days (72 hours). The crude extract was then filtered and evaporated to dryness. The dark paste was then stored in the refrigerator to be used later.
ANTIOXIDANT ASSAYS
Total phenol was by the method reported [4,5], total flavonoid [6], total antioxidant [7] iron (ll) complexation [8,9] DPPH quenching [10] nitric oxide quenching [11] copper coordination [12] hydroxyl radical sequestering [13]
Percentage yield = 17.36%
Table 1: Showing results of total phenol, flavonoid and total antioxidantin Sennaalata
Total phenol | Total flavonoid | Total antioxidant |
18.0 ± 2.82 mgGAE/g dry extract | 10.9 ± 0.0 mgQAE/g dry extract | 13.6 ± 0.08 mgAAE/g dry extract |
Values are mean ± S.D n = 3,GAE= Gallic Acid Equivalent, QAE= Quercetin equivalent, AAE= Ascorbic Acid Equivalent
Table 2: Showing results of Nitric oxide andDPPH scavenging in Sennaalata
| NO Scavenging | DPPH scavenging | ||
Conc. mg/ml | Senna alata | quercetin | Senna alata | Gallic acid |
0.1 | 6.49 ± 1.78
| 22.31± 0. 78
| 20.41±1.58
| 32.69 ± 1.74
|
0.2 | 16.32±1.09 | 25.79 ± 1.78
| 42.10±0.24
| 41.07 ± 0.14
|
0.4 | 30.16±1.41
| 46.09±1.35
| 64.29±1.41
| 56.31 ± 0.79
|
0.6 | 44.45 ± 0.61
| 68.88±0.08
| 76.70±1.51
| 73.77 ± 1.25
|
0.8 | 62.80 ± 2.79
| 71.38± 2.22
| 82.749±3.78 | 80.58 ± 0.04 |
1 | 74.73±1.41 | 89.32± 3.50 | 87.216±1.39 | 89.19 ± 0.04 |
Values are mean ± S.D n = 3
Nitric oxide (NO) and DPPH radicals were also sequestered by Senna alata extract is concentration dependent from 0.1 – 1 mg/ml concentration of Senna alata the percentage sequestration of NO and DPPH increases from 6.49 ± 1.78 % - 74.73±1.41 % and20.41±1.58 % - 87.216±1.39 % as shown in table 2 above.
Table 3: Hydroxyl radical scavenging in Sennaalata extract
Conc. (mg/ml) | Senna alata | Gallic acid |
0.1 | 24.93 ± 1.06 | 29.01 ± 0.05 |
0.2 | 35.119 ± 0.07 | 40.48 ± 0.14 |
0.4 | 58.825 ± 1.08 | 53.40 ± 0.11 |
0.6 | 71.83 ± 0.37 | 61.43 ± 0.14 |
0.8 | 83.09 ± 1.53 | 72.64 ± 0.79 |
1 | 92.84 ± 2.97 | 87.93 ± 0.31 |
Values are mean ± S.D n = 3
The hydroxyl radical is the most notorious ROS, in biological system. It is also produced by many metabolic reactions, Senna alata extract has the ability to quench the production of this radical. At concentrations of 0.1 mg/ml – 1mg/ml Senna alata scavenged the hydroxyl radical at 24.93 ± 1.06 % - 92.84 ± 2.97 % as shown on table 3 above. Gallic acid as standard also showed a higher level of scavenging of the hydroxyl radical.
Table 4 showing copper and iron chelation ability of Senna alata extract
| Copper chelating | Iron chelating | ||
Conc. mg/ml | Senna alata | Thiourea | Senna alata | EDTA |
0.1 | 16.60 ± 1.08 | 18.42 ± 2.61 | 19.82 ± 1.879 | 24.42 ± 11.99 |
0.2 | 35.95 ± 0.03 | 39.08 ± 0.77 | 38.23 ± 0.59 | 42.70 ± 1.74 |
0.4 | 49.16 ± 0.13 | 54.91 ± 1.97 | 47.50 ± 0.29 | 58.72 ± 2.09 |
0.6 | 57.28± 0.67 | 68.12 ± 0.20 | 57.42 ± 0.60 | 63.48 ± 0.26 |
0.8 | 77.64± 0.34 | 73.47 ± 0.20 | 72.87 ± 2.60 | 79.71 ±1.53 |
1.0 | 86.89 ± 0.23 | 81.69 ± 1.51 | 85.19 ± 1.63 | 86.62 ± 3.45 |
Values are mean ± S.D n = 3
Copper and iron coordination are important medical procedures, Senna alata chelation of copper is concentration dependent that is from 0.1 – 1 mg/ml concentration of Senna alata the percentage chelation of copper increases from 16.60±1.08 – 86.89±0.23 as shown in table 4 above. The standard copper chelating agent thiourea was marginally higher than the extract Senna alata. Senna alata chelation of iron is concentration dependent also that is from 0.1 – 1 mg/ml concentration of Senna alata the percentage chelation of iron increases from 19.82 ± 1.879 – 85.19 ± 1.63 as shown in table 4 above. The standard iron chelating agent EDTA was slightly higher than the extract Senna alata.
The findings of this investigation indicated that methanolic extract of Senna alata contains phytochemicals, that are radical scavenging and metal chelating properties which are present in different quantities with respect to the different phytocompounds.
The authors declare that they have no conflict of interest
No funding sources
The study was approved by the Niger Delta University, Bayelsa State, Nigeria.
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