Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea

In this study, major phytochemical compounds of <i>Vaccinium bracteatum</i> Thunb. (VB) aqueous leaf extract were isolated and analyzed using a HPLC-based method, followed by method validation in accordance with the International Conference on Harmonisation (ICH) guidelines for drug deve...

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Autores principales:	Seul-Gi Lee, Haeju Ko, Eun-Jin Choi, Dool-Ri Oh, Donghyuck Bae, Chulyung Choi
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Vaccinium bracteatum</i> Thunb. aqueous leaf extract isolation method validation reverse-phase HPLC vaccinoside Chemical technology TP1-1185 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/2604985449e9496db3e74bece24e1ba6
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spelling	oai:doaj.org-article:2604985449e9496db3e74bece24e1ba62021-11-25T18:49:55ZIsolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea10.3390/pr91118682227-9717https://doaj.org/article/2604985449e9496db3e74bece24e1ba62021-10-01T00:00:00Zhttps://www.mdpi.com/2227-9717/9/11/1868https://doaj.org/toc/2227-9717In this study, major phytochemical compounds of <i>Vaccinium bracteatum</i> Thunb. (VB) aqueous leaf extract were isolated and analyzed using a HPLC-based method, followed by method validation in accordance with the International Conference on Harmonisation (ICH) guidelines for drug development. Five major compounds were isolated in VB extract. Apart from vaccinoside, which had been the only compound isolated in VB extract to date, vanillic acid and protocatechuic acid were isolated for the first time. Isolation of orientin and isoorientin in the VB extract helped validate the reverse-phase analytical method. A new simple and rapid high-performance liquid chromatography (HPLC)-based method was developed for the validation of orientin and isoorientin in VB extract and was determinated according to the ICH guidelines. The analytical method was validated through a Waters Alliance HPLC System containing an e2695 separation module and a 2998 photodiode array (PDA) detector. The VB extract and solutions of orientin and isoorientin were analyzed using a reverse-phase Eclipse XDB-C18 column (4.6 × 250 mm ID, 5 µm, Waters), which was maintained at 30 °C. A mobile phase of methanol and 0.01% formic acid in water was used at a flow rate of 1.0 mL/min to achieve gradient elution. The linearity of the orientin and isoorientin was excellent results (R<sup>2</sup> ≥ 0.9999) in the concentration range of 1.0–50.0 μg/mL. Precision values ranged 98.55–101.70% and 98.70–101.18%, respectively. The intra-day and inter-day relative standard deviation (RSD) values of the orientin and isoorientin were all <2.0%. The average recoveries of orientin ranged 98.30–101.57%, whereas isoorientin ranged 97.81–102.14% with RSD values <2.0%. Quantitative analysis found that VB extract contained 2.90 mg/g of orientin and 3.45 mg/g of isoorientin.Seul-Gi LeeHaeju KoEun-Jin ChoiDool-Ri OhDonghyuck BaeChulyung ChoiMDPI AGarticle<i>Vaccinium bracteatum</i> Thunb.aqueous leaf extractisolationmethod validationreverse-phase HPLCvaccinosideChemical technologyTP1-1185ChemistryQD1-999ENProcesses, Vol 9, Iss 1868, p 1868 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Vaccinium bracteatum</i> Thunb. aqueous leaf extract isolation method validation reverse-phase HPLC vaccinoside Chemical technology TP1-1185 Chemistry QD1-999
spellingShingle	<i>Vaccinium bracteatum</i> Thunb. aqueous leaf extract isolation method validation reverse-phase HPLC vaccinoside Chemical technology TP1-1185 Chemistry QD1-999 Seul-Gi Lee Haeju Ko Eun-Jin Choi Dool-Ri Oh Donghyuck Bae Chulyung Choi Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea
description	In this study, major phytochemical compounds of <i>Vaccinium bracteatum</i> Thunb. (VB) aqueous leaf extract were isolated and analyzed using a HPLC-based method, followed by method validation in accordance with the International Conference on Harmonisation (ICH) guidelines for drug development. Five major compounds were isolated in VB extract. Apart from vaccinoside, which had been the only compound isolated in VB extract to date, vanillic acid and protocatechuic acid were isolated for the first time. Isolation of orientin and isoorientin in the VB extract helped validate the reverse-phase analytical method. A new simple and rapid high-performance liquid chromatography (HPLC)-based method was developed for the validation of orientin and isoorientin in VB extract and was determinated according to the ICH guidelines. The analytical method was validated through a Waters Alliance HPLC System containing an e2695 separation module and a 2998 photodiode array (PDA) detector. The VB extract and solutions of orientin and isoorientin were analyzed using a reverse-phase Eclipse XDB-C18 column (4.6 × 250 mm ID, 5 µm, Waters), which was maintained at 30 °C. A mobile phase of methanol and 0.01% formic acid in water was used at a flow rate of 1.0 mL/min to achieve gradient elution. The linearity of the orientin and isoorientin was excellent results (R<sup>2</sup> ≥ 0.9999) in the concentration range of 1.0–50.0 μg/mL. Precision values ranged 98.55–101.70% and 98.70–101.18%, respectively. The intra-day and inter-day relative standard deviation (RSD) values of the orientin and isoorientin were all <2.0%. The average recoveries of orientin ranged 98.30–101.57%, whereas isoorientin ranged 97.81–102.14% with RSD values <2.0%. Quantitative analysis found that VB extract contained 2.90 mg/g of orientin and 3.45 mg/g of isoorientin.
format	article
author	Seul-Gi Lee Haeju Ko Eun-Jin Choi Dool-Ri Oh Donghyuck Bae Chulyung Choi
author_facet	Seul-Gi Lee Haeju Ko Eun-Jin Choi Dool-Ri Oh Donghyuck Bae Chulyung Choi
author_sort	Seul-Gi Lee
title	Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea
title_short	Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea
title_full	Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea
title_fullStr	Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea
title_full_unstemmed	Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea
title_sort	isolation and analytical method validation for phytocomponents of aqueous leaf extracts from <i>vaccinium bracteatum</i> thunb. in korea
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/2604985449e9496db3e74bece24e1ba6
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Isolation and Analytical Method Validation for Phytocomponents of Aqueous Leaf Extracts from <i>Vaccinium bracteatum</i> Thunb. in Korea

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