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Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay

β-estradiol is an estrogen steroid hormone and acts as an estrogen receptor agonist. Radiolabeled β-estradiol is widely used as a radioligand for binding assays. In this present study, the synthesis of [(131)I]β-estradiol has been successfully carried out. Accordingly, the measurement of the radioch...

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Autores principales: Daruwati, Isti, Gwiharto, Abednego Kristande, Wongso, Hendris, Achmad, Tri Hanggono, Syaifudin, Mukh, Muchtaridi, Muchtaridi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Wolters Kluwer - Medknow 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10226700/
https://www.ncbi.nlm.nih.gov/pubmed/37255880
http://dx.doi.org/10.4103/japtr.japtr_624_22
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author Daruwati, Isti
Gwiharto, Abednego Kristande
Wongso, Hendris
Achmad, Tri Hanggono
Syaifudin, Mukh
Muchtaridi, Muchtaridi
author_facet Daruwati, Isti
Gwiharto, Abednego Kristande
Wongso, Hendris
Achmad, Tri Hanggono
Syaifudin, Mukh
Muchtaridi, Muchtaridi
author_sort Daruwati, Isti
collection PubMed
description β-estradiol is an estrogen steroid hormone and acts as an estrogen receptor agonist. Radiolabeled β-estradiol is widely used as a radioligand for binding assays. In this present study, the synthesis of [(131)I]β-estradiol has been successfully carried out. Accordingly, the measurement of the radiochemical purity (RCP) value and the presence of chemical impurities are needed. To validate the method for identifying the RCP and chemical impurities from [(131)I]β-estradiol using high-performance liquid chromatography (HPLC). The synthesis of [(131)I]β-estradiol was accomplished by a radioiodination reaction, and the RCP was determined by radio-HPLC. The method for β-estradiol measurement was validated by reversed-phase HPLC radio-analytical employing ultraviolet-visible (UV-Vis) and radioactive detector. The method for radio-HPLC analysis was validated and established using a C-18 column and MeCN: H(2)O (55:45 v/v) as the mobile phase. The following conditions were applied: a flow rate of 1.2 mL/min, isocratic, and a UV-Vis detector at 280 nm. The RCP of [(131)I]β-estradiol measured by thin-layer chromatography and radio-HPLC was 99.27% ± 1.25% and 95.75% ± 2.41%, respectively. The validation parameters were appropriate and met the requirements for acceptance. HPLC analysis was able to identify the presence of unlabeled estradiol (24.51%–27.29%) in the mixture of [(131)I]β-estradiol. As a result, purification using preparative HPLC or other methods will be required in future studies.
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spelling pubmed-102267002023-05-30 Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay Daruwati, Isti Gwiharto, Abednego Kristande Wongso, Hendris Achmad, Tri Hanggono Syaifudin, Mukh Muchtaridi, Muchtaridi J Adv Pharm Technol Res Original Article β-estradiol is an estrogen steroid hormone and acts as an estrogen receptor agonist. Radiolabeled β-estradiol is widely used as a radioligand for binding assays. In this present study, the synthesis of [(131)I]β-estradiol has been successfully carried out. Accordingly, the measurement of the radiochemical purity (RCP) value and the presence of chemical impurities are needed. To validate the method for identifying the RCP and chemical impurities from [(131)I]β-estradiol using high-performance liquid chromatography (HPLC). The synthesis of [(131)I]β-estradiol was accomplished by a radioiodination reaction, and the RCP was determined by radio-HPLC. The method for β-estradiol measurement was validated by reversed-phase HPLC radio-analytical employing ultraviolet-visible (UV-Vis) and radioactive detector. The method for radio-HPLC analysis was validated and established using a C-18 column and MeCN: H(2)O (55:45 v/v) as the mobile phase. The following conditions were applied: a flow rate of 1.2 mL/min, isocratic, and a UV-Vis detector at 280 nm. The RCP of [(131)I]β-estradiol measured by thin-layer chromatography and radio-HPLC was 99.27% ± 1.25% and 95.75% ± 2.41%, respectively. The validation parameters were appropriate and met the requirements for acceptance. HPLC analysis was able to identify the presence of unlabeled estradiol (24.51%–27.29%) in the mixture of [(131)I]β-estradiol. As a result, purification using preparative HPLC or other methods will be required in future studies. Wolters Kluwer - Medknow 2023 2023-04-13 /pmc/articles/PMC10226700/ /pubmed/37255880 http://dx.doi.org/10.4103/japtr.japtr_624_22 Text en Copyright: © 2023 Journal of Advanced Pharmaceutical Technology & Research https://creativecommons.org/licenses/by-nc-sa/4.0/This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.
spellingShingle Original Article
Daruwati, Isti
Gwiharto, Abednego Kristande
Wongso, Hendris
Achmad, Tri Hanggono
Syaifudin, Mukh
Muchtaridi, Muchtaridi
Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay
title Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay
title_full Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay
title_fullStr Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay
title_full_unstemmed Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay
title_short Method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)I]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay
title_sort method development, validation, and impurity measurement of β-estradiol from radiolabeled [(131)i]β-estradiol using radio-high-performance liquid chromatography for radioligand of saturation binding assay
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10226700/
https://www.ncbi.nlm.nih.gov/pubmed/37255880
http://dx.doi.org/10.4103/japtr.japtr_624_22
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