UPLC/Q-TOF-MS profiling of phenolics from <i>Canarium pimela</i> leaves and its vasorelaxant and antioxidant activities

Wu, Juan; Fang, Xiao'ai; Yuan, Yan; Dong, Yanfen; Liang, Yanling; Xie, Qingchun; Ban, Junfeng; Chen, Yanzhong; Lv, Zhufen

doi:10.1016/j.bjp.2017.10.005

Acessibilidade / Reportar erro

Brasil

Revista Brasileira de Farmacognosia

Español English

Brasil

Español English

sumário « anterior atual seguinte »

Sumário

Original Articles • Rev. bras. farmacogn. 27 (6) • Nov-Dec 2017 • https://doi.org/10.1016/j.bjp.2017.10.005 copy

UPLC/Q-TOF-MS profiling of phenolics from Canarium pimela leaves and its vasorelaxant and antioxidant activities

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

Canarium pimela K.D. Koenig, Burseraceae, have a long history of use in the Chinese traditional medicine treatment of various ailments including hypertension, and our research team has reported the anti-hypertensive activity and delineated the mechanism involved in the action. The following research aims to evaluate the vasorelaxant and antioxidant activities of ethanol extract from C. pimela leaves and to analyze its chemical composition by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) that may correlate with their pharmacological activities. The results showed that pre-incubation of aortic rings with the extract (0.3, 1, 3, 10, 30 and 100 mg/l) significantly inhibited the contractile response of the rings to norepinephrine-induced contraction (p < 0.01or p < 0.05). Crude ethanol extract and refined ethanol extract showed a highest inhibitory effect against 2,2dipheyl-2-picrylhydrazyl hydrate scavenging activity (IC₅₀ of crude ethanol extract = 15.42 ± 0.14 µg/ml and IC₅₀ of refined ethanol extract = 5.72 ± 0.31 µg/ml) and 2,2′-azinobis (3-ethyl-benzothiazoline-6-sulphonic acid ammonium salt) (ABTS (IC₅₀ of crude ethanol extract = 3.24 ± 0.18 µg/ml and IC₅₀ of refined ethanol extract = 1.88 ± 0.07 µg/ml) scavenging activity, which was considerably higher than that reported for butylated hydroxytoluene and lower of that measured for ascorbic acid. Moreover, its chemical composition was analyzed by UPLC/Q-TOF-MS. Sixteen compounds including nine flavonoids, four tannins, two phenolic acids and one dianthrone were identified for the first time as constituents of this species. And of this, six major phenolic components were simultaneous quantitative analysis by HPLC-UV, chlorogenic acid is the major compounds in C. pimela leaves. These results indicate that the phenolic-rich extract of C. pimela leaves is a promising natural pharmaceutical for combating hypertension and oxidative stress.

Keywords:
Phenolic; UPLC/Q-TOF-MS; Vasorelaxant; Antioxidant; Biological activity; HPLC-DAD

Compounds	Linearity ranges (mg/l)	Calibration curve^a a y = peak area, x = concentration in mg/l of extract.	(r²)	LOD^b b LOD, limit of detection. (mg/l)	LOQ^c c LOQ, limit of quantification. (mg/l)
Chlorogenic acid	57-570	y = 0.2402x + 0.3208	0.9998	0.13	0.48
Vitexin	2.5-25	y = 0.3108x - 0.1698	0.9996	0.52	1.89
Isovitexin	3.5-35	y = 0.6288x - 0.3924	0.9994	0.44	1.12
Hyperin	2.5-25	y = 0.642x - 0.3703	0.9995	0.41	1.02
Rutin	5.5-55	y = 0.5521x - 0.3689	0.9995	0.97	2.53
Quercetin	1-10	y = 0.9747x - 0.4955	0.9978	0.21	0.65

	150612		150820		150930
	Means ± SD (mg/g)	RSD (%)	Means ± SD (mg/g)	RSD (%)	Means ± SD (mg/g)	RSD (%)
Chlorogenic acid	13.39 ± 0.09	0.70	6.71 ± 0.09	1.39	13.42 ± 0.20	1.51
Vitexin	1.63 ± 0.03	1.80	0.56 ± 0.01	1.43	1.05 ± 0.01	1.06
Isovitexin	0.93 ± 0.01	0.66	0.47 ± 0.01	0.92	0.86 ± 0.02	1.76
Hyperin	1.04 ± 0.01	0.86	0.51 ± 0.01	1.35	0.88 ± 0.11	0.89
Rutin	2.04 ± 0.04	1.88	0.85 ± 0.02	1.91	1.62 ± 0.02	1.00
Quercetin	0.35 ± 0.01	1.49	0.16 ± 0.01	1.37	0.37 ± 0.01	0.65
Total (mg/g of leaf)	19.38 ± 2.28		9.26 ± 0.025		18.65 ± 0.06

Exstracts	Extraction yield (%)	Total phenolic content (mg GAE^a a GAE, gallic acid equivalents. /g of extract)	Flavonoids content (mg RE^d d RE, rutin equivalents. /g of extract)
CEE^b b CEE, crude ethanol extract.	20.95	259.1 ± 5.5	373.9 ± 6.56
REE^c c REE, refined ethanol extract.	13.4	415.6 ± 2.37**	824.1 ± 5.86^□□□

Number	Blank group	Isoproterenol group (ISO)		CPL leaf extract group
	Relaxation (%)	Final concentration lgC (g/l)	Relaxation (%)	CEE	REE
				Final concentration lgC (g/l)	Relaxation (%)
1	0.63 ± 1.47	-5.5	0.9 ± 2.03	-3.5	2.16 ± 2.81	2.98 ± 3.51
2	3.05 ± 2.33	-5.0	8.64 ± 7.45	-3.0	4.68 ± 5.61	5.89 ± 5.65
3	6.81 ± 3.70	-4.5	18.76 ± 9.1**	-2.5	14.25 ± 14.69	44.00 ±13.54**^□
4	10.57 ± 6.16	-4.0	34.72 ±16.7**	-2.0	65.77 ± 19.49**	91.25 ± 11.91**^□□
5	13.84 ± 6.42	-3.5	50.62 ± 14.18**	-1.5	90.38 ± 11.9**	100.18 ± 8.08**^□
6	19.23 ± 10.38	-3.0	54.16 ± 11.05**	-1.0	92.34 ± 9.78**	95.97 ± 7.88**

	IC₅₀ of DPPH^• (µg/ml)	IC₅₀ of ABTS^{• +} (µg/ml)	mg AAE^a a AAE, ascorbic acid equivalents. /g of extracts
Ascorbic acid	4.23 ± 0.23	1.78 ± 0.01	-
BHT	20.87 ± 0.40	-	-
CEE	15.42 ± 0.14	3.24 ± 0.18	274.39 ± 2.54
REE	5.72 ± 0.31***	1.88 ± 0.07^□□□	740.52 ± 40.65

Peak no.	R_t (min)	[M-H]^- (m/z)	Calculated (m/z)	Formula	Fragment ions (m/z)	Identified compounds	Ref/Std.
1	0.800	191.0576	191.0556	C₇H₁₂O₆	85,93,127	Quinic acid	^a a Ref.: Identified by comparison with references. Ref.
2	1.787	353.8078	353.1036	C₁₆H₁₈O₉	191,179,135	Chlorogenic acid	^b b Std.: Identified by comparison with a standard. Std.
3	2.635	633.0703	633.0728	C₂₇H₂₂O₁₈	463,301	Corilagin	^a a Ref.: Identified by comparison with references. Ref.
4	2.839	401.1617	401.1423	C₂₅H₂₂O₅	269,161,101	Unkonwn	-
5	3.112	505.1626	505.1506	C₃₀H₁₈O₈	343,293,169,151	Protohypericin	^a a Ref.: Identified by comparison with references. Ref.
6	3.672	451.1029	451.1501	C₂₄H₂₀O₉	341,217,177	Cinchonain I	^a a Ref.: Identified by comparison with references. Ref.
7	5.420	431.0978	431.1065	C₂₁H₂₀O₁₀	311,283	Vitexin	^b b Std.: Identified by comparison with a standard. Std.
8	6.148	431.0987	431.0992	C₂₁H₂₀O₁₀	341,311,283	Isovitexin	^b b Std.: Identified by comparison with a standard. Std.
9	6.494	463.0876	463.1071	C₂₁H₂₀O₁₂	300	Hyperoside	^b b Std.: Identified by comparison with a standard. Std.
10	6.721	609.1458	609.1792	C₂₇H₂₀O₁₆	300,179,151	Rutin	^b b Std.: Identified by comparison with a standard. Std.
11	7.403	433.0771	433.0820	C₂₀H₁₈O₁₁	300,271,255	Quercetin-3-O-arabinoside	^a a Ref.: Identified by comparison with references. Ref.
12	10.132	461.0773	461.0720	C₂₁H₁₈O₁₂	315,300	4'-O-methyellagic acid-3-O-α-l-rhamnopyranoside	^a a Ref.: Identified by comparison with references. Ref.
13	10.635	451.1029	451.1019	C₂₄H₂₀O₉	341,217,189	Cinchonain I isomer	^a a Ref.: Identified by comparison with references. Ref.
14	12.685	475.0904	475.0877	C₂₂H₂₀O₁₂	460,328,313	3,3'-Di-O-methylellagic acid-4-O-rhamnopyranoside	^a a Ref.: Identified by comparison with references. Ref.
15	14.503	301.0331	301.0292	C₁₅H₁₀O₇	179,151,121	Quercetin	^b b Std.: Identified by comparison with a standard. Std.
16	15.764	343.1539	343.1545	C₁₈H₁₆O₇	328,191,163	3,4,3'-Tri-O-methylellagic acid	^a a Ref.: Identified by comparison with references. Ref.
17	16.705	537.7937	537.1107	C₃₀H₁₈O₁₀	417,375	Amentoflavone	^a a Ref.: Identified by comparison with references. Ref.

Sociedade Brasileira de Farmacognosia Universidade Federal do Paraná, Laboratório de Farmacognosia, Rua Pref. Lothario Meissner, 632 - Jd. Botânico, 80210-170, Curitiba, PR, Brasil, Tel/FAX (41) 3360-4062 - Curitiba - PR - Brazil
E-mail: revista@sbfgnosia.org.br

Acompanhe os números deste periódico no seu leitor de RSS

[1] * Corresponding authors. xieqchun@163.com luzhufen@126.com