Phytochemical investigation and radical scavenging activities of Melia azedarach and its DNA protective effect in cultured lymphocytes

Figures & data

Figure 1. GC-MS analysis of ethanol leaf extract of Melia azedarach.

Table 1. Phytocomponents identified in the ethanol leaf extract of MA by GC-MS analysis.

S.No	RT	Name of the compound	Molecular formula	MW	Peak area %
1	6.76	Bicyclo[7.2.0]undec-4-ene, 4,11,11-trimethyl-8-methylene-,[1R-(1R*,4Z,9S*)]-	C15H24	204	0.58
2	8.39	1,6,10-Dodecatrien-3-ol, 3,7,11-trimethyl-,[S-(Z)]-	C15H26O	222	0.23
3	8.77	1H-3α,7-Methanoazulene, octahydro-1,4,9,9-tetramethyl-	C15H26	206	0.19
4	11.39	3,7,11,15-Tetramethyl-2-hexadecen-1-ol	C20H40O	296	3.51
5	11.48	2-Pentadecanone,6,10,14-trimethyl-	C18H36O	268	1.83
6	12.67	1-Hexadecen-3-ol,3,5,11,15-tetramethyl-	C20H40O	296	0.45
7	13.43	1H-Naptho[2,1-b]pyran,3-ethenyldodecahydro-3,4a,7,7,10a-pentamethyl-[3R-(3α,4αβ,6aα,10αβ,10bα)]-	C20H34O	290	1.19
8	13.67	1,6,10,14-Hexadecatetraen-3-ol,3,7,11,15-tetramethyl-,(E,E)-	C20H34O	290	1.19
9	14.81	Phytol	C20H40O	296	17.59
10	15.86	2,6,10-Dodecatrien-1-ol, 3,7,11-trimethyl-,acetate, (EE)-	C17H28O2	264	4.57
11	19.43	1-Naphthalenepropoanol,α-ethenyldecahydro-2-hydroxy-α,2,5,5,8a-pentamethyl-,[1R-[1α(R*), 2β,4aβ,8aα]-	C20H36O2	308	58.41
12	21.34	1,2-Benzenedicarboxylic acid, diisooctylester	C24H38O4	390	2.48
13	24.47	Squalene	C30H50	410	6.75

Table 2. IC₅₀ values of MA compared with that of standard ascorbic acid.

S.No	Radical scavenging assays	Compound/extract (µg/mL)	IC50 value (µg/mL)
1.	DPPH radical	Ascorbic acid	50.11 ± 0.50^b
		MA	30.55 ± 0.30^a
2.	Hydroxyl	Ascorbic acid	48.00 ± 0.48^b
		Melia azedarach	26.50 ± 0.26^a
3.	Superoxide anion	Ascorbic acid	34.50 ± 0.34^b
		Melia azedarach	30.00 ± 0.32^a
4.	Nitric oxide	Ascorbic acid	49.11 ± 0.49^a
		Melia azedarach	48.00 ± 0.48^a
5.	Evaluation of reducing power assay	Ascorbic acid	16.80 ± 0.16^a
		Melia azedarach	22.00 ± 0.22^b

Values are given as mean ± SD of six replicates in each group. Values are sharing a common superscript (a,b) differ significantly at p < 0.05 DMRT.

Figure 2. Hydroxyl radical scavenging effect of MA with different concentrations in comparison with standard ascorbic acid. Values are given as mean ± SD of six replicates in each group. Bar values are sharing a common superscript (a,b,c) differ significantly at p < 0.05 DMRT.

Figure 3. Superoxide radical scavenging effect of MA with different concentrations in comparison with standard ascorbic acid. Values are given as mean ± SD of six replicates in each group. Bar values are sharing a common superscript (a,b,c) differ significantly at p < 0.05 DMRT.

Figure 4. Nitric oxide radical scavenging effect of MA with different concentrations in comparison with standard ascorbic acid. Values are given as mean ± SD of six replicates in each group. Bar values are sharing a common superscript (a,b,c) differ significantly at p < 0.05 DMRT.

Figure 5. DPPH radical scavenging effect of MA with different concentrations in comparison with standard ascorbic acid. Values are given as mean ± SD of six replicates in each group. Bar values are sharing a common superscript (a,b,c) differ significantly at p < 0.05 DMRT.

Figure 6. Free radical scavenging activity of MA with different concentrations in comparison with standard ascorbic acid as measured in reducing power assay. Values are given as mean ± SD of six replicates in each group. Bar values are sharing a common superscript (a,b,c) differ significantly at p < 0.05 DMRT.

Figure 7. Protection against H₂O₂-induced lipid peroxidation in RBC cellular membrane by different concentrations of MA. Values are given as mean ± SD of six replicates in each group. Bar values are sharing a common superscript (a,b,c) differ significantly at p < 0.05 DMRT.

Figures 8. Changes in the levels of DNA damage (% DNA in tail, tail length, tail moment and Olive tail moment) in cultured lymphocytes on pretreatment with ethanol extract of MA. Values are given as mean ± SD of six replicates in each group. Bar values are sharing a common superscript (a,b,c) differ significantly at p < 0.05 DMRT.

Figure 9. Changes in the levels of DNA damage in normal, H₂O₂ and Melia azedarach pretreated lymphocytes.

Figure 10. Changes in the DNA fragmentation assay in cultured lymphocytes on pretreatment with ethanol extract of MA. Values are given as mean ± SD of six replicates in each group. Lane (L1)-1000bP ladder DNA; L2-Normal lymphocytes; L3-Normal + MA (60 µg/mL); L4-H₂O₂ (500 µM); L5-H₂O₂ + MA (60 µg/mL).