Phytochemical analysis, antiproliferative and antioxidant activities of Chrozophora tinctoria: a natural dye plant

Figures & data

Figure 1. HPLC-TOF/MS chromatograms of the extracts from C. tinctoria: (a) ethyl acetate extract, (b) n-BuOH extract, (c) methanol-chloroform extract, (d) water extract.

Figure 2. The main compounds from the extracts determined by HPLC/TOF-MS (a) ethyl acetate extract, (b) n-BuOH extract, (c) methanol-chloroform extract, (d) water extract.

Table 1. Quantification of phenolic compounds of the extracts.

	Water	EA	n-BuOH	MeCh
Gallic acid	13.98	68.92	31.12	1.08
Caffeic acid	ND	3.91	ND	ND
p-Coumaric acid	ND	0.15	ND	ND
Rosmarinic acid	0.20	2.19	0.69	0.09
Chicoric acid	ND	43.93	0.08	ND
Apigenin-7-glucoside	ND	133.34	5.57	13.68
Quercetin	ND	0.44	0.12	ND
Naringenin	ND	ND	ND	ND
Kaempferol	ND	ND	ND	ND
Gentisic acid	2.60	12.51	24.02	ND
4-Hydroxy benzoic acid	0.88	268.20	ND	3.76
Chlorogenic acid	0.67	ND	ND	ND
Vanillic acid	6.44	23.53	3.00	ND
Ferulic acid	ND	43.51	0.35	ND
Salicylic acid	ND	12.33	1.99	ND
Catechin	0.34	0.33	1.02	ND
Protocateuchic acid	ND	3.71	1.62	ND
4-hydroxybenzaldehyde	ND	2.16	ND	ND
Rutin	ND	3.78	2.27	ND
Ellagic acid	0.15	49.59	2.39	ND
Hesperidin	ND	1.88	ND	ND
Protocateuchic acid ethyl ester	ND	ND	ND	ND
Resveratrol	1.46	ND	ND	ND

ND: Not determined.

Figure 3. Antiproliferative effects of the ethyl acetate and butanol extracts (a) and active fractions (b) from C. tinctoria.

Table 2. The antiproliferative effects of C. tinctoria EA extract and obtained fractions at the tested concentrations.

Concentration (μg/mL)	EA extract	CT/E-F1	CT/E-F2	CT/E-F3	CT/E-F4	CT/E-F5	CT/E-F6	CT/E-F7	CT/E-F8	5-FU
100	15.07 ± 0.00	38.47 ± 0.39	−0.43 ± 0.04	9.49 ± 0.66	11.66 ± 0.26	15.32 ± 0.24	92.47 ± 0.71	6.04 ± 0.30	−1.65 ± 0.04	83.20 ± 0.33
75	−3.12 ± 0.22	1.40 ± 0.33	−0.73 ± 0.05	3.58 ± 0.09	−1.24 ± 0.05	9.81 ± 0.05	67.28 ± 0.09	4.43 ± 0.02	−1.72 ± 0.03	81.86 ± 0.43
50	−1.55 ± 0.02	3.64 ± 0.78	−0.33 ± 0.03	0.40 ± 0.05	0.74 ± 0.01	−0.99 ± 0.14	16.56 ± 0.24	−0.92 ± 0.02	−0.28 ± 0.01	81.41 ± 0.13
40	−2.41 ± 0.18	3.93 ± 0.34	−0.46 ± 0.06	0.04 ± 0.00	−1.38 ± 0.04	−1.95 ± 0.12	1.25 ± 0.06	−1.66 ± 0.01	−2.34 ± 0.04	80.82 ± 0.45
30	−1.54 ± 0.16	−0.54 ± 0.05	−0.49 ± 0.02	−0.31 ± 0.01	−0.84 ± 0.02	−1.59 ± 0.05	−0.74 ± 0.01	−1.17 ± 0.03	−0.01 ± 0.02	80.98 ± 0.09
20	−2.65 ± 0.12	−2.03 ± 0.12	−1.41 ± 0.22	−1.47 ± 0.00	−1.39 ± 0.06	−1.67 ± 0.16	−1.63 ± 0.02	−2.21 ± 0.01	−1.17 ± 0.03	81.66 ± 0.07
10	−1.61 ± 0.22	−0.74 ± 0.03	−0.80 ± 0.12	−0.23 ± 0.02	−0.52 ± 0.03	−0.67 ± 0.03	−0.99 ± 0.01	−0.75 ± 0.05	−1.37 ± 0.05	80.16 ± 0.60
5	−2.41 ± 0.06	0.65 ± 0.08	−1.13 ± 0.04	−0.93 ± 0.00	−0.13 ± 0.01	−0.40 ± 0.06	−0.56 ± 0.02	−0.65 ± 0.02	−1.14 ± 0.08	77.58 ± 0.72
	CT/E − F9	CT/E − F10	CT/E − F11	CT/E − F12	CT/E − F13	CT/E − F15	CT/E − F16	CT/E − F17	CT/E − F18	CT/E − F19
100	−1.60 ± 0.14	−1.14 ± 0.15	0.89 ± 0.04	−1.31 ± 0.18	−1.18 ± 0.18	0.84 ± 0.03	−0.14 ± 0.01	−0.24 ± 0.00	6.09 ± 0.05	−0.40 ± 0.01
75	−2.09 ± 0.08	−0.58 ± 0.05	−1.40 ± 0.04	−2.01 ± 0.04	−1.31 ± 0.04	−1.85 ± 0.05	−2.93 ± 0.40	−2.17 ± 0.06	−1.62 ± 0.28	−0.16 ± 0.01
50	−1.54 ± 0.08	−1.09 ± 0.18	−0.27 ± 0.01	−1.55 ± 0.36	−0.81 ± 0.06	−1.80 ± 0.07	−2.22 ± 0.16	−1.43 ± 0.34	−1.96 ± 0.02	−0.40 ± 0.08
40	−2.42 ± 0.07	−1.92 ± 0.32	−1.10 ± 0.03	−1.10 ± 0.28	−1.93 ± 0.08	−2.43 ± 0.06	−2.78 ± 0.24	−2.05 ± 0.24	−1.53 ± 0.48	−0.53 ± 0.04
30	−2.18 ± 0.20	−1.72 ± 0.12	−1.36 ± 0.04	−1.62 ± 0.14	−1.02 ± 0.12	−1.92 ± 0.08	−2.12 ± 0.66	−2.16 ± 0.16	−1.55 ± 0.42	−0.31 ± 0.08
20	−3.22 ± 0.08	−2.26 ± 0.40	−2.39 ± 0.03	−2.71 ± 0.36	−2.09 ± 0.18	−2.50 ± 0.03	−2.47 ± 0.16	−3.15 ± 0.16	−2.67 ± 0.08	−1.26 ± 0.01
10	−0.92 ± 0.07	0.23 ± 0.01	−0.93 ± 0.02	−1.54 ± 0.02	−1.12 ± 0.02	−1.43 ± 0.08	−1.73 ± 0.36	−2.03 ± 0.42	−1.85 ± 0.02	−0.83 ± 0.02
5	−1.54 ± 0.08	0.17 ± 0.02	−1.95 ± 0.03	−2.16 ± 0.66	−1.69 ± 0.05	−1.66 ± 0.05	−1.58 ± 0.06	−1.72 ± 0.18	−1.61 ± 0.14	−0.54 ± 0.06

Table 4. DPPH radical scavenging and antiproliferative effects of major phenolic compounds of EA extract.

Main phenolic compounds	DPPH radical scavenging IC50 (μg/mL)	Antiproliferative effect IC50 (μg/mL)
Gallic acid	<5	<5
Apigenin 7-glucoside	134.7 ± 1.2	>100
4-Hydroxybenzoic acid	>150	>100

Figure 4. DPPH radical scavenging of the EA extract, fractions, and BHT at the tested concentrations.

Table 3. Antioxidant activities of the fractions and ethyl acetate extract from C. tinctoria*.

Material	DPPH IC50 μg/mL	Total phenolic content μg GAE/mg fraction
EA	27.3 ± 0.4^c	115.3 ± 0.4^d
CT/E-F1	63.1 ± 4.2^h	74.8 ± 3.2^j
CT/E-F2	14.0 ± 0.0^a	186.4 ± 1.1^a
CT/E-F3	31.9 ± 0.3^d,e	100.7 ± 1.4^f.g
CT/E-F4	16.2 ± 0.5^a	138.9 ± 0.7^b
CT/E-F5	18.7 ± 0.0^a	125.5 ± 0.5^c
CT/E-F6	17.3 ± 0.1^a	126.4 ± 0.4^c
CT/E-F7	18.6 ± 0.4^a	126.4 ± 0.4^c
CT/E-F8	17.8 ± 0.4^a	127.6 ± 0.5^c
CT/E-F9	>150	9.2 ± 0.6^l
CT/E-F10	>150	25.9 ± 0.6^k
CT/E-F11	32.8 ± 0.7^e	98.3 ± 0.2^g
CT/E-F12	43.1 ± 1.8^g	88.2 ± 0.3^h
CT/E-F13	61.2 ± 2.3^h	73.8 ± 0.6^j
CT/E-F14	39.2 ± 0.8^f	99.7 ± 0.6^g
CT/E-F15	32.0 ± 1.4^d,e	100.9 ± 1.1^g
CT/E-F16	27.3 ± 2.9^c	113.2 ± 0.3^d
CT/E-F17	28.8 ± 0.8^c,d	108.7 ± 0.2^e
CT/E-F18	30.8 ± 0.2^d,e	100.9 ± 0.7^f
CT/E-F19	48.8 ± 0.4^g	85.0 ± 0.7^i
BHT	23.3 ± 0.6^b	NS

* Values represent averages ± standard deviations for triplicate experiments. Values in the same column with different superscripts are significantly (p < 0.01) different, NS: Not Studied.