Applicability of an HPLC method for analysis of alcoholic and glycolic Brazilian green-propolis extracts

Vecchi, Camila Felix; Santos, Rafaela Said dos; Bruschi, Marcos Luciano

doi:10.1590/s2175-97902023e21972

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Brazilian Journal of Pharmaceutical Sciences

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Article • Braz. J. Pharm. Sci. 59 • 2023 • https://doi.org/10.1590/s2175-97902023e21972 copy

Applicability of an HPLC method for analysis of alcoholic and glycolic Brazilian green-propolis extracts

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Brazilian green propolis has been widely used in food and pharmaceutical products due to its valuable source of phenolic compounds and versatile biological activities. The development and validation of analytical methods are extremely useful for the characterization and quality control of products containing propolis. Therefore, the aim of this study was to optimize, validate and investigate the applicability of a reversed-phase HPLC method for analysis of different types of Brazilian green propolis extracts (glycolic and ethanolic). The method showed to be selective for the propolis phenolic markers. The analysis of variance and residues demonstrated that the method had significant linear regression, without lack of fit. It was also a precise, accurate and robust method, which was of utmost importance to analyze both glycolic and ethanolic extracts and at different concentrations. Moreover, as these products can display most complex matrices to analyze, a valid HPLC method can also prove to be specific and versatile.

Keywords:
Propolis; High-performance liquid chromatography (HPLC); Glycolic extract; Ethanolic extract; Phytochemicals

	Sum of squares	Degree of freedom	Mean square	F-value	F-critical_95%
(p-Coumaric acid)
Regression	36994810508	1	36994810508	6597.41	4.10
Residual	213084212.7	38	5607479.282
Lack-of-fit	49422772	213084212.76	8237129	1.61	2.40
Pureerror	163661440	32	5114420	1.61	2.40
(Chrysin)
Regression	2.28602x10¹¹	1	2.28602x10¹¹	15003.79	4.05
Residual	716105313.8	47	15236283.27
Lack-of-fit	152099858	6	25349976	1.64	2.30
Pure error	741236241	48	15442422	1.64	2.30

PRECISION
	Repeatability			Intermediate precision
p-coumaric acid (µg/mL)	Intraday	RSD (%)	Interday	RSD (%)	Interanalist	RSD (%)
0.03	0.0192 ± 0.0003	1.32	0.0198 ± 0.0010	4.83	0.0329 ± 0.0015	4.69
0.10	0.0867 ± 0.0021	2.41	0.0903 ± 0.0031	3.46	0.0938 ± 0.0045	4.85
0.30	0.3179 ± 0.0051	1.61	0.2842 ± 0.0104	3.66	0.2780 ± 0.0135	4.87
Chrysin (µg/mL)	Intraday	RSD (%)	Interday	RSD (%)	Interanalist	RSD (%)
0.30	0.3653 ± 0.0064	1.75	0.3200 ± 0.0072	2.24	0.3263 ± 0.0126	3.85
2.00	1.9032 ± 0.0619	3.25	1.9603 ± 0.0347	1.77	1.9281 ± 0.0775	4.02
4.00	4.0239 ± 0.1597	3.97	4.0930 ± 0.0642	1.57	4.0708 ± 0.1975	4.85

ACCURACY
p-coumaric acid (µg/mL)	p-coumaric acid concentration found (µg/mL)	Recovery (%)	Mean recovery (%)
0.40	0.3924 ± 0.0098	96.41 ± 2.36
0.50	0.49936 ± 0.0125	98.15 ± 2.83	98.22 ± 0.24
0.60	0.6163 ± 0.0158	100.12 ± 2.58
Chrysin (µg/mL)	Chrysin concentration found (µg/mL)	Recovery (%)	Mean recovery (%)
0.30	0.3177 ± 0.0152	103.91 ± 5.36
0.90	0.8982 ± 0.0457	97.93 ± 5.30	98.17 ± 1.39
3.00	2.8332 ± 0.0915	92.65 ± 2.93

p-Coumaric acid (µg/mL)	305 nm	310 nm	315 nm	RSD (%)
0.40	0.3834± 0.0028	0.3914± 0.0026	0.37599 ± 0.0021	2.01
0.50	0.4903 ± 0.0109	0.4990 ± 0.0113	0.4797 ± 0.0108	1.97
0.60	0.5980 ± 0.0144	0.6088 ± 0.0144	0.5860 ± 0.0137	1.91
Chrysin (µg/mL)	305 nm	310 nm	315 nm	RSD (%)
0.30	0.3172 ± 0.0048	0.3273 ± 0.0056	0.3325 ±0.0059	3.03
0.90	0.8949 ± 0.0362	0.9156 ± 0.0336	0.9383 ± 0.0378	2.56
2.00	1.9030 ± 0.0515	1.9775 ± 0.0560	2.0187 ± 0.0574	3.09

Propolis extract	Content (%, w/v)
Propolis extract	p-coumaric acid	Chrysin
EE10%	0.0026 ± 0.00001	0.0041 ± 0.00003
EE30%	0.0121 ± 0.0.00002	0.0180 ± 0.0002
GE10%	0.0021 ± 0.00001	0.0009 ± 0.00002
GE30%	0.0096 ± 0.0001	0.0014 ± 0.00002

Universidade de São Paulo, Faculdade de Ciências Farmacêuticas Av. Prof. Lineu Prestes, n. 580, 05508-000 S. Paulo/SP Brasil, Tel.: (55 11) 3091-3824 - São Paulo - SP - Brazil
E-mail: bjps@usp.br

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[1] *Correspondence: M. L. Bruschi. Departamento de Farmácia. Centro de Ciências da Saúde. Universidade Estadual de Maringá. Av. Colombo, 5790, Bloco K68, Sala 05. CEP 87020-900, Maringá, PR, Brasil. Phone: +55 44 3011-5998 / 4870. E-mail: mlbruschi@uem.br.