Podophyllum hexandrum ameliorates endosulfan-induced genotoxicity and mutagenicity in freshwater cyprinid fish crucian carp

Figures & data

Table 1. Frequency profile of CA induced by endosulfan and P. hexandrum extracts separately followed by their combination for different time intervals to evaluate the antimutagenicity in C. carassius.

			Classical aberrations						Non-classical aberrations
Experiments	Treatment	TMS	Csb	Ctb	Frg	Scu	Dic	Mla	Stp	Cmt	Total aberrations,Mean (%) ± S.D.
48	Cont. 1	105	1	1	–	–	–	–	1	–	2.85 ± 0.21^2
	Cont. 2	102	1	1	–	–	–	–	–	1	2.94 ± 0.16^2
	ES	117	2	2	2	–	1	–	2	–	7.69 ± 0.37^Bb
	HEPH	114	–	1	1	–	–	–	1	1	3.50 ± 0.18^2
	CEPH	108	1	2	–	–	–	–	–	1	3.70 ± 0.23^2
	EEPH	111	1	1	1	–	–	–	1	–	3.60 ± 0.18^2
	MEPH	100	2	1	–	–	–	–	–	–	3.00 ± 0.22^2
	AEPH	115	1	1	–	–	–	–	1	1	3.47 ± 0.18^2
	ES + HEPH	110	2	1	1	1	1	1	1	–	7.27 ± 0.29^Bb
	ES + CEPH	113	2	2	1	–	1	1	2	–	7.96 ± 0.35^Bb
	ES + EEPH	109	1	2	1	–	–	–	1	1	5.50 ± 0.26^Bb2
	ES + MEPH	103	1	2	1	–	–	–	1	–	4.85 ± 0.25^Bb2
	ES + AEPH	107	1	2	1	1	1	–	1	–	6.54 ± 0.28^Bb
72	Cont. 1	104	1	1	–	–	–	–	1	–	2.88 ± 0.16^2
	Cont. 2	101	1	–	1	–	–	–	1	–	2.97 ± 0.17^2
	ES	110	2	2	1	1	1	–	2	1	9.09 ± 0.37^Bb
	HEPH	116	1	1	1	–	–	–	–	1	3.44 ± 0.18^2
	CEPH	114	1	1	2	–	–	–	–	–	3.50 ± 0.22^2
	EEPH	113	–	1	2	–	–	–	–	1	3.53 ± 0.22^2
	MEPH	109	1	1	1	–	–	–	1	–	3.66 ± 0.18^2
	AEPH	100	–	1	1	–	–	–	1	–	3.00 ± 0.17^2
	ES + HEPH	106	2	2	1	1	1	1	1	–	8.41 ± 0.33^Bb
	ES + CEPH	112	1	2	2	1	1	2	1	–	8.92 ± 0.36^Bb
	ES + EEPH	117	1	2	2	1	–	–	–	1	5.98 ± 0.30^Bb2
	ES + MEPH	115	2	1	1	–	–	1	–	1	5.21 ± 0.25^Bb2
	ES + AEPH	111	1	1	2	2	1	2	–	–	8.10 ± 0.36^Bb
96	Cont. 1	119	2	1	–	–	–	–	1	–	3.36 ± 0.22^2
	Cont. 2	108	–	–	2	–	–	–	1	1	3.70 ± 0.23^2
	ES	107	3	2	1	1	2	2	1	1	12.14 ± 0.47^B
	HEPH	108	1	1	1	–	–	–	–	1	3.70 ± 0.18^2
	CEPH	102	–	1	2	–	–	–	1	–	3.92 ± 0.24^2
	EEPH	107	2	1	–	–	–	–	–	1	3.73 ± 0.23^2
	MEPH	105	1	–	1	–	–	–	1	1	3.80 ± 0.19^2
	AEPH	101	1	2	1	–	–	–	–	–	3.96 ± 0.24^2
	ES + HEPH	110	2	2	2	2	1	2	1	1	11.81 ± 0.44^Bb
	ES + CEPH	109	2	1	2	2	2	2	1	1	11.92 ± 0.44^Bb
	ES + EEPH	116	1	2	2	1	1	1	–	–	6.89 ± 0.31^Bb2
	ES + MEPH	113	1	1	1	1	1	1	–	–	6.19 ± 0.24^Bb2
	ES + AEPH	104	2	2	2	1	1	1	1	–	9.61 ± 0.38^Bb2

Exp: exposure time in hours; TMS: total metaphasic plates studied; Csb: chromosome break; Ctb: chromatid break; Frg: fragment; Scu: sister chromatid union; Dic: dicentric; Mla; multiple aberrations; Stp: stickiness and pulverization; Cmt: c metaphase; Cont. 1: negative control (tap water); Cont. 2: solvent control; ES: endosulfan; HEPH; CEPH; EEPH; MEPH and AEPH represent the hexane, chloroform, ethyl acetate, methanol and aqueous extract of P. hexandrum, respectively. Values with different upper and lower letter superscripts differ significantly (^Aap < .05 = significant; ^Bbp < .01 = highly significant; ^Ccp < .001 = extremely significant) from the control 1 and 2, respectively (Newman–Keuls and Dunnett’s multiple comparison test), whereas values with different numeric superscripts differ significantly (¹p < .05 = significant; ²p < .01 = highly significant; ³p < .001 = extremely significant) from the endosulfan group (Dunnett’s multiple comparison test).

Table 2. Frequency profile of CA induced alone by endosulfan and in combination with the variable concentrations of the active extracts of P. hexandrum for different time intervals to evaluate the concentration-dependent antimutagenic response in C. carassius.

			Classical aberrations						Non-classical aberrations
Exp.	Treatment	TMS	Csb	Ctb	Frg	Scu	Dic	Mla	Stp	Cmt	Total aberrationsMean (%)± S.D.
48	Cont. 1	105	1	1	–	–	–	–	1	–	2.85 ± 0.21^2
	Cont. 2	102	1	1	–	–	–	–	–	1	2.94 ± 0.16^2
	Endosulfan	117	2	2	1	1	1	–	2	–	7.69 ± 0.37^Bb
	EE-T1	111	2	1	2	1	–	–	–	1	6.30 ± 0.30^Bb
	EE-T2	104	1	2	1	1	1	–	–	–	5.76 ± 0.27^Bb2
	EE-T3	109	1	2	1	–	–	–	1	1	5.50 ± 0.26^Bb2
	ME-T1	110	–	1	2	1	1	1	–	–	5.55 ± 0.26^Bb2
	ME-T2	108	1	1	1	–	–	1	–	1	4.62 ± 0.21^Aa2
	ME-T3	103	1	2	1	–	–	–	1	–	4.85 ± 0.25^Bb2
72	Cont. 1	104	1	1	–	–	–	–	1	–	2.88 ± 0.16^2
	Cont. 2	101	1	–	1	–	–	–	1	–	2.97 ± 0.17^2
	Endosulfan	110	2	2	1	1	1	–	2	1	9.09 ± 0.37^Bb
	EE-T1	116	1	2	1	1	1	2	–	–	6.89 ± 0.31^Bb2
	EE-T2	111	1	1	1	1	1	2	–	–	6.30 ± 0.27^Bb2
	EE-T3	112	1	2	2	1	–	–	–	1	5.98 ± 0.30^Bb2
	ME-T1	113	1	2	1	1	1	1	–	–	6.21 ± 0.27^Bb2
	ME-T2	119	2	1	1	1	1	–	–	–	5.04 ± 0.25^Ba2
	ME-T3	115	2	1	1	–	–	1	–	1	5.21 ± 0.25^Bb2
96	Cont. 1	119	2	1	–	–	–	–	1	–	3.36 ± 0.22^2
	Cont. 2	108	–	–	2	–	–	–	1	1	3.70 ± 0.23^2
	Endosulfan	107	3	2	1	1	2	2	1	1	12.14 ± 0.47^Bb
	EE-T1	117	2	2	2	1	1	2	–	–	8.54 ± 0.38^Bb2
	EE-T2	101	1	1	2	1	1	1	1	–	7.92 ± 0.30^Bb2
	EE-T3	116	1	2	2	1	1	1	–	–	6.89 ± 0.31^Bb2
	ME-T1	107	1	1	2	1	1	2	–	–	7.47 ± 0.32^Bb2
	ME-T2	118	1	1	1	1	1	1	1	–	5.93 ± 0.23^Bb2
	ME-T3	113	1	1	1	1	1	1	–	–	6.19 ± 0.24^Bb2

Exp: exposure time in hours; TMS: total metaphasic plates studied; Csb: chromosome break; Ctb: chromatid break; Frg: fragment; Scu: sister chromatid union; Dic: dicentric; Mla: multiple aberrations; Stp: stickiness and pulverization; Cmt: c metaphase; Cont. 1: negative control (tap water); Cont. 2: solvent control; EE-T1: ethyl acetate extract-treatment 1(0.05 mg/L endosulfan +5 mg/L EE); EE-T2: ethyl acetate extract-treatment 2 (0.05 mg/L endosulfan +10 mg/L EE); EE-T3: ethyl acetate extract-treatment 3 (0.05 mg/L endosulfan +15 mg/L EE); ME-T1: methanol extract treatment 1(0.05 mg/L endosulfan +5 mg/L ME); ME-T2: methanol extract treatment 2 (0.05 mg/L endosulfan +10 mg/L ME); ME-T3: methanol extract treatment 3 (0.05 mg/L endosulfan +15 mg/L ME).

Table 3. Frequency profile of CA induced by endosulfan alone and in combination with column eluted fractions of active P. hexandrum extracts for different time intervals to evaluate the antimutagenicity in C. carassius.

			Classical aberrations						Non-classical aberrations
Exp.	Treatment	TMS	Csb	Ctb	Frg	Scu	Dic	Mla	Stp	Cmt	Total aberrations mean (%)± S.D.
48	Cont. 1	105	1	1	–	–	–	–	1	–	2.85 ± 0.21^2
	Cont. 2	102	1	1	–	–	–	–	–	1	2.94 ± 0.16^2
	Endosulfan	117	2	2	1	1	1	–	2	–	7.69 ± 0.37^Bb
	EE-F1	103	1	1	–	1	1	2	–	–	5.82 ± 0.27^Bb2
	EE-F2	106	1	1	–	1	1	1	–	1	5.66 ± 0.23^Bb2
	EE-F3	113	1	1	2	1	1	–	–	–	5.30 ± 0.26^Bb2
	EE-F4	101	1	–	1	–	–	1	1	1	4.95 ± 0.21^Bb2
	EE-F5	107	1	2	1	1	1	–	–	–	5.60 ± 0.26^Bb2
	ME-F1	116	1	1	1	1	1	1	–	–	5.17 ± 0.22^Bb2
	ME-F2	118	1	1	2	–	–	1	–	–	4.23 ± 0.24^2
	ME-F3	113	2	1	1	–	–	1	–	–	4.42 ± 0.24^A2
	ME-F4	114	1	–	1	1	1	–	1	1	5.26 ± 0.22^Bb2
	ME-F5	115	1	2	2	1	1	–	–	–	6.08 ± 0.30^Bb1
72	Cont. 1	104	1	1	–	–	–	–	1	–	2.88 ± 0.16^2
	Cont. 2	101	1	–	1	–	–	–	1	–	2.97 ± 0.17^2
	Endosulfan	110	2	2	1	1	1	–	2	1	9.09 ± 0.37^Bb
	EE-F1	120	1	1	1	2	2	–	1	–	6.66 ± 0.31^Bb2
	EE-F2	100	1	1	1	1	1	1	–	–	6.00 ± 0.23^Bb2
	EE-F3	119	2	1	2	–	–	1	1	–	5.88 ± 0.29^Bb2
	EE-F4	102	1	–	1	–	–	1	1	1	4.90 ± 0.21^Bb2
	EE-F5	108	–	1	2	–	–	2	1	1	6.48 ± 0.31^Bb2
	ME-F1	120	1	1	2	–	–	2	1	–	5.83 ± 0.29^Bb2
	ME-F2	114	1	1	1	–	–	1	–	1	4.38 ± 0.20^2
	ME-F3	107	2	1	1	1	1	–	–	–	5.60 ± 0.26^Bb2
	ME-F4	109	1	1	1	–	–	1	1	1	5.50 ± 0.22^Bb2
	ME-F5	103	1	2	1	1	1	1	–	–	6.79 ± 0.28^Bb2
96	Cont. 1	119	2	1	–	–	–	–	1	–	3.36 ± 0.22^2
	Cont. 2	108	–	–	2	–	–	–	1	1	3.70 ± 0.23^2
	Endosulfan	107	3	2	1	1	2	2	1	1	12.14 ± 0.47^Bb
	EE-F1	109	1	1	2	1	1	1	1	1	8.25 ± 0.33^Bb2
	EE-F2	111	2	1	1	1	1	1	1	–	7.20 ± 0.29^Bb2
	EE-F3	112	1	1	1	2	2	1	–	–	7.14 ± 0.32^Bb2
	EE-F4	105	1	1	2	–	–	1	1	–	5.71 ± 0.27^Bb2
	EE-F5	106	1	2	1	1	1	–	1	1	7.54 ± 0.29^Bb2
	ME-F1	113	–	2	3	–	–	2	–	1	7.07 ± 0.39^Bb2
	ME-F2	103	–	1	2	–	–	1	1	–	4.85 ± 0.25^2
	ME-F3	104	2	1	1	–	–	1	1	1	6.73 ± 0.28^Bb2
	ME-F4	108	2	2	1	–	–	2	–	1	7.40 ± 0.35^Bb2
	ME-F5	116	1	2	1	2	2	–	1	–	7.75 ± 0.35^Bb2

Exp: exposure time in hours; TMS: total metaphasic plates studied; Csb: chromosome break; Ctb: chromatid break; Frg: fragment; Scu: sister chromatid union; Dic: dicentric; Mla: multiple aberrations; Stp: stickiness and pulverization; Cmt: c metaphase; Cont. 1: negative control (tap water); Cont. 2: solvent control; EE-F1,2,3,4, and 5 designate the ethyl acetate extract fractions 1,2,3,4, and 5 (15 mg/L each), respectively, ME-F1,2,3,4, and 5 represent the methanol extract fractions 1,2,3,4, and 5 (10 mg/L), respectively.

Table 4. Frequency profiles of micronuclei induced alone by endosulfan and P. hexandrum extracts followed by their simultaneous exposure for different time intervals to evaluate antimutagenicity in C. carassius.

			Number of MN
Exp.	Treatment	Total cells scored	1	2	3	Total no. of MN	Frequency of MN mean (%)± SD
48	Cont. 1	11,800	28	–	–	28	0.23 ± 0.13^2
	Cont. 2	11,320	28	1	–	30	0.26 ± 0.13^2
	ES	12,000	321	21	6	381	3.17 ± 0.83^Bb
	HEPH	11,490	30	3	–	36	0.31 ± 0.14^2
	CEPH	11,730	34	4	–	42	0.35 ± 0.15^2
	EEPH	11,980	31	1	–	33	0.27 ± 0.14^2
	MEPH	11,670	31	–	–	31	0.26 ± 0.15^2
	AEPH	11,560	33	1	–	35	0.30 ± 0.16^2
	ES + HEPH	11,330	334	13	5	375	3.30 ± 0.92^Bb
	ES + CEPH	11,840	329	15	6	377	3.18 ± 0.87^Bb
	ES + EEPH	11,648	177	11	2	205	1.75 ± 0.47^Bb2
	ES + MEPH	11,594	135	9	2	159	1.37 ± 0.36^Aa2
	ES + AEPH	11,210	321	8	1	340	3.03 ± 0.90^Bb
72	Cont. 1	11,550	23	–	–	23	0.19 ± 0.21^2
	Cont. 2	11,000	30	–	–	30	0.27 ± 0.11^2
	Endosulfan	11,250	320	35	4	402	3.57 ± 0.89^Bb
	HEPH	11,990	37	3	–	43	0.35 ± 0.17^2
	CEPH	11,100	38	5	–	48	0.43 ± 0.18^2
	EEPH	11,388	35	1	–	37	0.32 ± 0.17^2
	MEPH	11,715	37	1	–	39	0.33 ± 0.17^2
	AEPH	11,845	34	–	–	34	0.29 ± 0.16^2
	ES + HEPH	11,465	337	17	8	395	3.44 ± 0.92^Bb
	ES + CEPH	11,635	331	23	11	410	3.52 ± 0.89^Bb
	ES + EEPH	11,585	145	10	2	171	1.47 ± 0.39^Ba2
	ES + MEPH	11,560	120	16	4	164	1.41 ± 0.32^Ba2
	ES + AEPH	12,000	271	15	8	325	2.70 ± 0.70^Bb
96	Cont. 1	11,980	34	1	–	36	0.30 ± 0.16^2
	Cont. 2	11,220	36	1	–	38	0.33 ± 0.18^2
	Endosulfan	11,760	413	116	23	714	6.07 ± 1.11^Bb
	HEPH	11,068	34	5	–	44	0.39 ± 0.15^2
	CEPH	11,609	41	3	–	47	0.40 ± 0.19^2
	EEPH	11,872	38	1	–	40	0.33 ± 0.18^2
	MEPH	11,039	45	–	–	45	0.40 ± 0.23^2
	AEPH	11,317	45	2	–	49	0.43 ± 0.22^2
	ES + HEPH	11,000	421	89	27	680	6.18 ± 1.20^Bb
	ES + CEPH	11,427	440	99	19	695	6.08 ± 1.22^Bb
	ES + EEPH	11,005	178	46	15	315	2.86 ± 0.52^Bb2
	ES + MEPH	11,740	197	33	9	290	2.47 ± 0.52^Bb2
	ES + AEPH	12,000	386	68	21	585	4.87 ± 1.01^Bb1

Exp: exposure time in hours; Cont. 1: negative control (tap water); Cont. 2: solvent control; ES: endosulfan (0.05 mg/L); HEPH, CEPH, EEPH, MEPH and AEPH represent the hexane, chloroform, ethyl acetate, methanol and aqueous extract (15 mg/L each) of P. hexandrum, respectively.

Table 5. Frequency profiles of micronuclei induced alone by endosulfan and in combination with the variable concentrations of the active extracts of P. hexandrum for different time intervals to evaluate the concentration-dependent antimutagenic response in C. carassius.

			Number of MN
Exp.	Treatment	Total cells scored	1	2	3	Total no. of MN	Frequency of MN mean (%)± SD
48	Cont. 1	11,800	28	–	–	28	0.23 ± 0.13^2
	Cont. 2	11,320	28	1	–	30	0.26 ± 0.13^2
	Endosulfan	12,000	321	21	6	381	3.17 ± 0.83^Bb
	EE-T1	11,905	203	11	2	231	1.94 ± 0.53^Bb
	EE-T2	11,730	192	8	3	217	1.84 ± 0.51^Bb1
	EE-T3	11,648	177	11	2	205	1.75 ± 0.47^Aa1
	ME-T1	11,560	118	23	7	185	1.60 ± 0.31^Aa2
	ME-T2	11,670	109	17	4	155	1.32 ± 0.29^2
	ME-T3	11,594	135	9	2	159	1.37 ± 0.36^2
72	Cont. 1	11,550	23	–	–	23	0.19 ± 0.21^2
	Cont. 2	11,000	30	–	–	30	0.27 ± 0.11^2
	Endosulfan	11,250	320	35	4	402	3.57 ± 0.89^Bb
	EE-T1	11,490	200	11	7	243	2.11 ± 0.96^Bb1
	EE-T2	11,730	121	29	5	194	1.65 ± 0.52^Aa2
	EE-T3	11,585	145	10	2	171	1.47 ± 0.69^A2
	ME-T1	11,670	144	19	3	191	1.63 ± 0.66^Aa2
	ME-T2	11,160	109	12	2	139	1.24 ± 0.52^2
	ME-T3	11,560	120	16	4	164	1.41 ± 0.55^2
96	Cont. 1	11,980	34	1	–	36	0.30 ± 0.16^2
	Cont. 2	11,220	36	1	–	38	0.33 ± 0.18^2
	Endosulfan	11,760	413	116	23	714	6.07 ± 1.11^Bb
	EE-T1	11,490	163	77	16	365	3.17 ± 0.64^Bb2
	EE-T2	11,730	126	91	26	386	3.29 ± 0.43^Bb2
	EE-T3	11,005	183	51	10	315	2.86 ± 0.80^Bb2
	ME-T1	11,670	127	61	21	312	2.67 ± 0.45^Bb2
	ME-T2	11,560	123	49	17	272	2.35 ± 0.47^Bb2
	ME-T3	11,740	197	33	9	290	2.47 ± 0.87^Bb2

Exp: exposure time in hours; Cont. 1: negative control (tap water); Cont. 2: solvent control; EE-T1: ethyl acetate extract-treatment 1(0.05 mg/L endosulfan +5 mg/L EE); EE-T2: ethyl acetate extract-treatment 2 (0.05 mg/L endosulfan +10 mg/L EE); EE-T3: ethyl acetate extract-treatment 3 (0.05 mg/L endosulfan +15 mg/L EE); ME-T1: methanol extract-treatment 1(0.05 mg/L endosulfan +5 mg/L ME); ME-T2: methanol extract-treatment 2 (0.05 mg/L endosulfan +10 mg/L ME); ME-T3: methanol extract-treatment 3 (0.05 mg/L endosulfan +15 mg/L ME).

Table 6. Frequency profiles of micronuclei induced by endosulfan alone and in combination with column eluted fractions of active P. hexandrum extracts for different time intervals to evaluate the antimutagenicity in C. carassius.

			Number of MN
Exp.	Treatment	Total cells scored	1	2	3	Total no. of MN	Frequency of MN mean (%)± SD
48	Cont. 1	11,800	28	–	–	28	0.23 ± 0.13^2
	Cont. 2	11,320	28	1	–	30	0.26 ± 0.13^2
	Endosulfan	12,000	321	21	6	381	3.17 ± 0.83^Bb
	EE-F1	11,209	181	7	3	204	1.81 ± 0.90^Aa1
	EE-F2	11,200	136	13	5	177	1.58 ± 0.65^Aa2
	EE-F3	11,977	140	11	4	174	1.45 ± 0.63^2
	EE-F4	11,743	138	8	2	160	1.36 ± 0.65^2
	EE-F5	11,037	184	11	4	218	1.97 ± 0.92^Bb
	ME-F1	11,410	113	25	6	181	1.58 ± 0.50^Aa2
	ME-F2	11,080	90	6	3	111	1.00 ± 0.44^2
	ME-F3	11,635	112	9	3	139	1.19 ± 0.52^2
	ME-F4	11,094	133	15	9	190	1.71 ± 0.62^Aa1
	ME-F5	11,595	229	18	5	280	2.41 ± 1.08^Bb
72	Cont. 1	11,550	23	–	–	23	0.19 ± 0.21^2
	Cont. 2	11,000	30	–	–	30	0.27 ± 0.11^2
	Endosulfan	11,250	320	35	4	402	3.57 ± 0.89^Bb
	EE-F1	11,715	181	27	6	253	2.15 ± 0.81^Bb1
	EE-F2	11,940	159	23	4	217	1.81 ± 0.70^Ba2
	EE-F3	11,107	138	19	4	188	1.69 ± 0.66^Aa2
	EE-F4	11,333	101	14	3	138	1.21 ± 0.47^2
	EE-F5	12,000	168	31	7	251	2.09 ± 0.72^Bb1
	ME-F1	11,820	172	12	3	205	1.73 ± 0.80^Aa2
	ME-F2	11,440	83	6	1	98	0.85 ± 0.40^2
	ME-F3	11,255	128	11	2	156	1.38 ± 0.62^2
	ME-F4	11060	132	30	5	207	1.87 ± 0.60^Bb2
	ME-F5	11,677	214	29	8	296	2.53 ± 0.97^Bb
96	Cont. 1	11,980	34	1	–	36	0.30 ± 0.16^2
	Cont. 2	11,220	36	1	–	38	0.33 ± 0.18^2
	Endosulfan	11,760	413	116	23	714	6.07 ± 1.11^Bb
	EE-F1	11,300	197	101	18	453	4.00 ± 0.79^Bb2
	EE-F2	11,785	176	76	14	370	3.13 ± 0.69^Bb2
	EE-F3	11,950	169	71	11	344	2.87 ± 0.66^Bb2
	EE-F4	11,563	133	35	6	221	1.91 ± 0.57^Aa2
	EE-F5	11,110	166	79	16	372	3.34 ± 0.70^Bb2
	ME-F1	11,270	155	65	10	315	2.79 ± 0.64^Bb2
	ME-F2	11,830	121	7	3	144	1.21 ± 0.56^2
	ME-F3	11,615	185	53	8	315	2.71 ± 0.79^Bb2
	ME-F4	11,404	183	56	9	322	2.82 ± 0.78^Bb2
	ME-F5	11,909	216	98	17	463	3.88 ± 0.84^Bb2

Exp: exposure time in hours; Cont. 1: negative control (tap water); Cont. 2: solvent control; EE-F1,2,3,4 and 5 designate the ethyl acetate extract fraction 1,2,3,4, and 5 (15 mg/L each), respectively; ME-F1,2,3,4, and 5 represent the methanol extract fraction 1,2,3,4, and 5 (10 mg/L), respectively.

Figure 1. Antioxidant activity of active fractions of P. hexandrum expressed as rutin equivalents by the DPPH method.

Figure 2. TIC chromatogram of the methanol (ME-F2) and ethyl acetate (EE-F4) fraction of P. hexandrum. Peaks (A) 1: 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one; 2: 2-furancarboxaldehyde, 5-(hydroxymethyl); 3: hexadecanoic acid, methyl ester. (B) 1: n-hexadecanoic acid; 2: (3β)-stigmast-5-en-3-ol; 3: (3β)-stigmasta-5,22-dien-3-ol acetate; 4: deoxy-podophyllotoxin; 5: podophyllotoxin; 6: epiisopodophyllotoxin-acetate; X: minor fractions (detailed in results).