Analysis of flavonoids in <i>Rubus erythrocladus</i> and   <i>Morus nigra</i> leaves extracts by liquid chromatography and capillary electrophoresis

Tallini, Luciana R.; Pedrazza, Graziele P.R.; Bordignon, Sérgio A. de L.; Costa, Ana C.O.; Steppe, Martin; Fuentefria, Alexandre; Zuanazzi, José A.S.

doi:10.1016/j.bjp.2015.04.003

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Original Articles • Rev. bras. farmacogn. 25 (3) • May-Jun 2015 • https://doi.org/10.1016/j.bjp.2015.04.003 copy

Analysis of flavonoids in Rubus erythrocladus and Morus nigra leaves extracts by liquid chromatography and capillary electrophoresis

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

This study uses high performance liquid chromatography and capillary electrophoresis as analytical tools to evaluate flavonoids in hydrolyzed leaves extracts of Rubus erythrocladus Mart., Rosaceae, and Morus nigra L., Moraceae. For phytochemical analysis, the extracts were prepared by acid hydrolysis and ultrasonic bath and analyzed by high performance liquid chromatography using an ultraviolet detector and by capillary electrophoresis equipped with a diode-array detector. Quercetin and kaempferol were identified in these extracts. The analytical methods developed were validated and applied. Quercetin and kaempferol were quantified in R. erythrocladus, with 848.43 ± 66.68 μg g^-1 and 304.35 ± 17.29 μg g^-1, respectively, by HPLC-UV and quercetin, 836.37 ± 149.43 μg g^-1, by CE-DAD. In M. nigra the quantifications of quercetin and kaempferol were 2323.90 ± 145.35 μg g^-1 and 1446.36 ± 59.00 μg g^-1, respectively, by HPLC-UV and, 2552.82 ± 275.30 μg g^-1 and 1188.67 ± 99.21 μg g^-1, respectively, by CE-DAD. The extracts were also analyzed by ultra-performance liquid chromatography coupled with a diode-array detector and mass spectrometer (MS), UPLC-DAD/MS.

Keywords
HPLC-UV; UPLC-DAD/MS; CE-DAD; Rubus erythrocladus ; Morus nigra ; Flavonoids

	Flavonoid	Linear equation	R ²	LOD (μg ml^-1)	LOQ (μg ml^-1)	Precision intraday	Precision interday	Mean recovery (%) ± RSD%
HPLC	Quercetin	y = 83082 x + 42,829	0.9996	0.9519	2.8844	0.30	1.69	95.35 ± 2.88
	Kaempferol	y = 69225 x + 56,605	0.9995	1.0519	3.1876	0.23	2.02	95.76 ± 2.77
CE	Quercetin	y = 0.0176 x + 0.0276	0.9979	5.3043	16.0737	2.48	7.76	104.56 ± 1.77
	Kaempferol	y = 0.0203 x + 0.005	0.9987	4.1887	12.6931	3.68	8.44	104.31 ± 1.66

HPLC parameter	Quercetin	Kaempferol	CE Parameter	Quercetin	Kaempferol
Flow 0.55 ml min^-1	109.40 ± 0.31	109.40 ± 0.45	25.5 °C	97.63 ± 0.26	106.27 ± 0.07
Flow 0.65 ml min^-1	92.35 ± 0.26	92.38 ± 0.02	24.5 °C	98.39 ± 0.63	106.37 ± 0.71
0.005% TFA	95.89 ± 7.70	93.50 ± 11.58	2 s	95.76 ± 1.87	107.73 ± 0.14
0.015% TFA	101.57 ± 0.13	100.14 ± 0.39	4 s	97.94 ± 0.25	103.19 ± 4.12
0.04%TFA	102.96 ± 0.61	100.89 ± 0.69	5.0%	99.30 ± 3.71	98.73 ± 2.41
0.12% TFA	104.90 ± 0.16	104.28 ± 0.05	15.0%	98.77 ± 1.43	104.76 ± 0.16
DAD	94.27 ± 0.54	94.29 ± 0.71	–	–	–

	Flavonoid	R. erythrocladus Mean (μg g^-1) ± RSD%	M. nigra Mean (μg g^-1) ± RSD%
HPLC	Quercetin	848.43 ± 10.48	2323.90 ± 7.28
	Kaempferol	304.35 ± 6.83	1446.36 ± 6.72
CE	Quercetin	836.37 ± 9.36	2552.82 ± 7.96
	Kaempferol	N.Q.	1188.67 ± 9.20

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[1] * Corresponding author. E-mail: zuanazzi@ufrgs.br (J.A.S. Zuanazzi).