Chemical composition, antioxidant activity and antitumor activity of tumorous stem mustard leaf and stem extracts

Figures & data

Figure 1. Plant of tumorous stem mustard (Brassica juncea var. tumida Tsen et Lee) (a); and total polyphenols and total flavonoids contents in extracts from tumorous stem mustard leaf and stem, respectively (b). TSMLE, tumorous stem mustard leaf extract; TSMSE, tumorous stem mustard stem extract. *p< 0.05, ***p< 0.001, as compared to the TSMLE.

Figura 1. Planta de mostaza de tallo tumoral (Brassica juncea var. tumida Tsen et Lee) (A); y contenido de polifenoles totales y flavonoides totales en extractos de hoja y tallo de mostaza de tallo tumoral, respectivamente (B). TSMLE, extracto de hoja de mostaza de tallo tumoral; TSMSE, extracto de tallo de mostaza de tallo tumoral. *p< 0.05, ***p<0.001, en comparación con el TSMLE.

Figure 2. Phytochemicals profile of tumorous stem mustard leaf and stem extracts. The percentage of phytochemicals in tumorous stem mustard leaf and stem extracts (a). The main individual organic acids, polyphenols and other compounds (b); and vitamins and nucleotide derivates (c) in tumorous stem mustard leaf and stem extracts. blue: organic acids; orange: others; purple: polyphenols; green: vitamins; black: nucleotide and its derivates.

Figura 2. Perfil de fitoquímicos de extractos de hoja y tallo de mostaza de tallo tumoral. Porcentaje de fitoquímicos en extractos de hojas y tallos de mostaza de tallo tumoral (A). Principales ácidos orgánicos individuales, polifenoles y otros compuestos (B); y vitaminas y derivados de nucleótidos (C) en extractos de hojas y tallos de mostaza de tallo tumoral. Azul: ácidos orgánicos; naranja: otros; morado: polifenoles; verde: vitaminas; negro: nucleótido y sus derivados.

Table 1. The phytochemicals that differently existed between tumorous stem mustard stem extracts (TSMSE) and tumorous stem mustard leaf extracts (TSMLE).

Tabla 1. Fitoquímicos presentes de manera diferente en los extractos de tallos de mostaza de tallo tumoral (TSMSE) y los extractos de hojas de mostaza de tallo tumoral (TSMLE).

Class	Compounds	Ion mode	Q1 (Da)	Q3 (Da)	TSMLE (%)	TSMSE (%)
Anthocyanins	Cyanidin 3-O-glucoside (Kuromanin)	+	449.100	287.300	1.06850	0.00070
Anthocyanins	Cyanidin 3,5-O-diglucoside (Cyanin)	+	611.000	287.700	0.84239	0.00062
Anthocyanins	Delphinidin 3-O-glucoside (Mirtillin)	+	465.100	303.100	0.14498	0.00016
Anthocyanins	Cyanidin O-acetylhexoside	-	489.100	285.300	0.02381	0.00004
Anthocyanins	Cyanidin	+	287.000	231.600	0.04478	0.00023
Anthocyanins	Delphinidin	+	303.000	149.300	0.00256	0.00002
Anthocyanins	Rosinidin O-hexoside	+	477.100	315.600	0.00993	0.00022
Benzoic acid derivatives	4-Hydroxy-3,5-diisopropylbenzaldehyde	+	207.200	165.500	0.00410	0.00007
Catechin derivatives	Gallocatechin-gallocatechin	+	611.200	611.100	0.85569	0.00089
Coumarins	6,7-dihydroxycoumarin 7-O-quinic acid	-	351.100	132.600	0.19818	0.00026
Coumarins	N-sinapoyl hydroxycoumarin	+	369.119	207.500	0.76702	0.00344
Coumarins	Esculin (6,7-Dihydroxycoumarin-6-glucoside)	-	339.100	177.100	0.03636	0.00033
Coumarins	O-Feruloyl 4-hydroxylcoumarin	+	339.100	177.500	1.13057	0.01313
Flavanone	Hesperetin 5-O-glucoside	-	463.132	301.100	0.86456	0.00183
Flavanone	Naringenin O-malonylhexoside	+	521.000	317.600	0.02842	0.00049
Flavone	Selgin O-malonylhexoside	+	565.000	317.800	0.10020	0.00005
Flavone	Luteolin O-hexosyl-O-hexosyl-O-hexoside	-	771.100	609.500	1.59167	0.00229
Flavone	Luteolin 7-O-glucoside (Cynaroside)	+	449.100	287.100	1.02860	0.00439
Flavone	Luteolin 3‘,7-di-O-glucoside	+	611.100	449.400	0.00736	0.00005
Flavone	Chrysoeriol 5-O-hexoside	-	461.100	285.200	0.09887	0.00139
Flavone C-glycosides	8-C-hexosyl-hesperetin O-hexoside	+	627.100	430.000	0.20616	0.00017
Flavone C-glycosides	C-hexosyl-luteolin O-sinapic acid	+	655.200	655.000	0.01215	0.00002
Flavone C-glycosides	Eriodictiol C-hexosyl-O-hexoside	+	613.100	300.300	0.05409	0.00019
Flavone C-glycosides	6-C-hexosyl-luteolin O-hexoside	+	611.100	329.000	0.00406	0.00002
Flavone C-glycosides	Chrysoeriol C-pentosyl-O-hexosyl-O-hexoside	-	755.100	311.400	0.00829	0.00007
Flavone C-glycosides	Eriodictiol 6-C-hexoside 8-C-hexoside-O-hexoside	+	775.100	774.900	0.00100	0.00001
Flavone C-glycosides	C-hexosyl-luteolin O-p-coumaroylhexoside	+	757.200	757.000	0.01818	0.00021
Flavonol	Isorhamnetin O-acetyl-hexoside	-	519.100	315.300	0.10064	0.00013
Flavonol	Kaempferol 3-O-glucoside (Astragalin)	-	447.101	287.100	1.19708	0.00219
Flavonol	Quercetin 3-O-glucoside (Isotrifoliin)	+	465.000	302.800	0.86899	0.00221
Flavonol	methylQuercetin O-hexoside	+	479.000	317.300	2.07050	0.00900
Flavonol	Dihydroquercetin (Taxifolin)	-	303.058	125.100	0.00802	0.00005
Flavonol	Isorhamnetin 5-O-hexoside	+	479.200	317.800	2.15474	0.01270
Flavonol	Kaempferol 3-O-galactoside (Trifolin)	+	449.000	286.900	1.11284	0.00707
Flavonol	Isorhamnetin	-	315.058	300.100	0.08424	0.00078
Flavonol	Isorhamnetin O-hexoside	+	479.200	318.000	1.32122	0.01221
Hydroxycinnamoyl derivatives	Caffeic acid O-glucoside	-	341.000	179.200	0.06251	0.00010
Organic acids	Glutaric acid	-	131.042	86.900	0.28242	0.00398
Organic acids	ethylmalonate	-	131.040	86.900	0.01033	0.00017
Organic acids	2–furanoic acid	-	111.016	79.900	0.21813	0.00464
Quinate and its derivatives	5-O-p-coumaroyl quinic acid O-hexoside	-	499.100	163.200	0.02066	0.00001
Quinate and its derivatives	1-O-Caffeoyl quinic acid	-	353.100	191.100	0.39459	0.00022
Quinate and its derivatives	1-O-p-Coumaroyl quinic acid	-	337.100	155.800	0.09931	0.00011
Quinate and its derivatives	3-O-p-Coumaroyl quinic acid	-	337.100	190.900	0.35203	0.00091
Quinate and its derivatives	Homovanilloyl quinic acid	-	355.100	181.200	0.00156	0.00001
Quinate and its derivatives	1-O-Feruloyl quinic acid	+	369.100	207.500	0.08468	0.00073
Quinate and its derivatives	Neochlorogenic acid (5-O-Caffeoylquinic acid)	-	353.000	191.100	0.19685	0.00337
Quinate and its derivatives	3-O-p-Coumaroyl shikimic acid	-	319.000	145.300	0.02008	0.00042
Quinate and its derivatives	3-O-Feruloyl quinic acid	+	369.100	177.500	0.00745	0.00016
Quinate and its derivatives	Chlorogenic acid methyl ester	-	367.000	179.100	0.06695	0.00158

Q1: precursor ion, Q3: qualitative ion.

Q1: precursor de ion, Q3: ion cualitativo.

Figure 3. In vitro antioxidant effects of tumorous stem mustard leaf and stem extracts. (a) ABTS⁺-scavenging assay; (b) DPPH·-scavenging assay; (c) total reducing power; (d) ferric reducing antioxidant power (FRAP). Tea polyphenols extract (pure >97%) was used as a positive control. Results were expressed as the mean ± SD (n = 3).

Figura 3. Efectos antioxidantes in vitro de extractos de hojas y tallos de mostaza de tallo tumoral. (A) ensayo de eliminación de ABTS +; (B) ensayo de eliminación de DPPH ·; (C) poder reductor total; (D) Poder de reducción antioxidante del [ion] férrico (FRAP). Se utilizó el extracto de polifenoles de té (puro> 97%) como control positivo. Los resultados se expresan como la media ± desviación estándar (n = 3).

Figure 4. Effects of tumorous stem mustard leaf and stem extracts on cellular viability and ROS generation in human lung cancer A549 cells. Cells were treated with or without different concentrations of TSMLE and TSMSE (0, 30, 60, 120, 240, and 480 μg/mL) for 24 h (a). Cells were treated with or without different concentrations of TSMLE for 12, 24 and 48 h (b). Data were mean values ± SD of 10 duplications. ^a-c Mean values with different alphabetical letters indicate significant difference (p< 0.0001, ^c’ p< 0.001) at the same treatment concentration. The fluorescence intensity of DHE (O₂⁻) was measured using DHE probes (c).

Figura 4. Efectos de extractos de hoja y tallo de mostaza de tallo tumoral en la viabilidad celular y la generación de ROS en células de cáncer de pulmón humano A549. Las células se trataron con o sin concentraciones diferentes de TSMLE y TSMSE (0, 30, 60, 120, 240 y 480 μg/ml) durante 24 h (A). Las células se trataron con o sin concentraciones diferentes de TSMLE durante 12, 24 y 48 h (B). Los datos son valores medios ± DE de 10 duplicaciones. a-c Los valores medios con diferentes letras alfabéticas indican una diferencia significativa (p< 0.0001, ^c’ p< 0.001) en la misma concentración de tratamiento. La intensidad de fluorescencia de DHE (O2-) se midió utilizando sondas DHE (C).