Development and validation of an HPLC method for the determination of epicatechin in Maytenus ilicifolia (Schrad.) Planch., Celastraceae

Abstract

RESUMO: "Desenvolvimento e validação de um método de CLAE para a determinação da epicatequina em Maytenus ilicifolia (Schrad.) Planch., Celastraceae". Um método simples, reprodutível e eficiente de cromatografia líquida de alta eficiência (CLAE) foi desenvolvido. Água (0,05% TFA):acetonitrila (0,05% TFA) foi utilizado como fase móvel em um sistema de gradiente para a determinação da epicatequina (EP) em folhas de Maytenus ilicifolia (Schrad.) Planch., Celastraceae. A análise foi realizada utilizando coluna RP C-18 (5 μm) como fase estacionária, com vazão de 0,8 mL/min, e comprimento de onda de 210 nm para a detecção e determinação. Os principais parâmetros de validação do método foram determinados. A curva analítica apresentouse linear no intervalo de 10-120 µg/mL (EP). O coeficiente de correlação da análise de regressão linear foi de 0,9988, o limite de detecção e o limite de quantificação foram de 28,61 e 86,77 µg/ mL, respectivamente. O conteúdo do EP foi determinado com sucesso, com boa reprodutibilidade e recuperação. Recuperação da EP foi 99,32%. O método foi aplicado com sucesso na determinação da epicatequina em folhas de M. ilicifolia. A avaliação interlaboratorial demonstrou a reprodutibilidade do método com desvio padrão relativo de 14,62%. Unitermos: Maytenus ilicifolia, CLAE, epicatequina, desenvolvimento de método analítico, controle de qualidadeABSTRACT: A simple, reproducible and efficient high-performance liquid chromatography (HPLC) method was developed. Water (0.05% TFA):acetonitrile (0.05% TFA) was used as the mobile phase in a gradient system for the determination of epicatechin (EP) in leaves of Maytenus ilicifolia (Schrad.) Planch. The analysis was performed using an RP C-18 column (5 µm) as the stationary phase, with a flow rate of 0.8 mL/min, at a wavelength of 210 nm for detection and determination. The main validation parameters of the method were also determined. The calibration curve was found to be linear, with a range of 10-120 µg/mL (EP). The correlation coefficient of the linear regression analysis was within 0.9988, and the detection and quantification limits were 28.61 and 86.77 µg/mL, respectively. The content of EP was successfully determined, with satisfactory reproducibility and recovery. Recovery of the EP was 99.32%. The method was successfully applied to the determination of epicatechin in leaves of M. ilicifolia. The interlaboratorial evaluation showed the reproducibility of the method with a relative standard deviation of 14.62%.

Application and Analysis of the Folin Ciocalteu Method for the Determination of the Total Phenolic Content from Limonium Brasiliense L.

“…Although quantitative determination of polyphenols is hampered by their structural complexity and diversity, several methods have used to determine polyphenols in plant extracts [4,5,6,7]. Assuming that quantification of individual polyphenols does not adequately reveal the proportion of polymeric procyanidins, then spectrophotometry in the ultraviolet region may be a useful tool to help resolve this problem [8,9].…”

A quantitative validated method using liquid chromatography and chemometric analysis for evaluation of raw material oF Maytenus ilicifolia (Schrad.) Planch., Celastraceae

“…For this purpose, a quantitative method was developed employing epicatechin as chemical marker of the M. ilicifolia species, as recommended by the Brazilian Pharmacopoeia. 7,18,30 To achieve this goal, a LC-DAD system was applied making the proposed method adequate to be used in the quality control of herbal medicines made from M. ilicifolia.…”

“…7,17 For this reason, many Brazilian pharmaceutical industries and shops, produce and market phytopharmaceuticals containing this plant. 18 It is well accepted that for evaluating the quality of crude herbal material and phytomedicines it is necessary to standardise the content of the active principles. However, when the active principles are not known, a compound, which is the most representative of the chemical composition of the extract, may be selected as the marker (chemical marker).…”

Development and validation of a high performance liquid chromatography-diode array detection (HPLC-DAD) method for the quantification of rutin and isoquercetin in Morus nigra L. (Moraceae)

“…In 2009, the Ministry of Health disclosed the National List of Medicinal Plants of Interest (RENISUS), to the Unified Health System (SUS) being the Morus genus among the plants included in this list. It is noteworthy that the availability of validated assay methods is an important part of the quality control of these products, and such assays are required by the Brazilian health authorities for registration of phytomedicines (Lopes et al, 2010).…”