Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometry
2-Methoxyestradiol (2ME) is involved in the pathogenesis of preeclampsia and antitumor activity. In addition to its low concentration in healthy human serum, presence of isomers makes quantification of 2ME for clinical research and laboratory medicine difficult. The objective of this study was to de...
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Elsevier
2025-04-01
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| Series: | Practical Laboratory Medicine |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352551724000933 |
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| author | Koji Takahashi Masaki Takiwaki Seketsu Fukuzawa Yoshikuni Kikutani Kentaro Abe Tatsuya Higashi Hironori Kobayashi |
| author_facet | Koji Takahashi Masaki Takiwaki Seketsu Fukuzawa Yoshikuni Kikutani Kentaro Abe Tatsuya Higashi Hironori Kobayashi |
| author_sort | Koji Takahashi |
| collection | DOAJ |
| description | 2-Methoxyestradiol (2ME) is involved in the pathogenesis of preeclampsia and antitumor activity. In addition to its low concentration in healthy human serum, presence of isomers makes quantification of 2ME for clinical research and laboratory medicine difficult. The objective of this study was to develop a highly sensitive and accurate method for quantifying 2ME using LC-MS/MS combined with derivatization with 1-(2,4-dinitro-5-fluorophenyl)-4,4-dimethylpiperazinium iodide (MPDNP-F). This approach significantly enhanced the detectability of 2ME in positive electrospray ionization-tandem mass spectrometry (ESI-MS/MS) and enabled the chromatographic separation of 2ME from isomeric metabolites possessing a methoxy group, including 4-methoxyestradiol, 3-O-methyl 2-hydroxyestradiol, and 3-O-methyl 4-hydroxyestradiol (3M4OH). Although the derivatized 2ME and 3M4OH were closely eluted under the optimized LC conditions, the different fragmentation patterns of these isomers during MS/MS allowed their distinction. The lower limit of quantification for 2ME was 2.5 pg/mL, indicating a satisfactory sensitivity. These findings demonstrated that this LC-MS/MS method combined with the MPDNP-F derivatization can provide accurate and highly sensitive quantification of 2ME. |
| format | Article |
| id | doaj-art-baff64e6e980406ca47ffc67274475b0 |
| institution | Kabale University |
| issn | 2352-5517 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Practical Laboratory Medicine |
| spelling | doaj-art-baff64e6e980406ca47ffc67274475b02025-08-20T03:42:41ZengElsevierPractical Laboratory Medicine2352-55172025-04-0144e0044710.1016/j.plabm.2024.e00447Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometryKoji Takahashi0Masaki Takiwaki1Seketsu Fukuzawa2Yoshikuni Kikutani3Kentaro Abe4Tatsuya Higashi5Hironori Kobayashi6Medical Equipment Business Operations, JEOL Ltd., Tokyo, JapanMedical Equipment Business Operations, JEOL Ltd., Tokyo, Japan; Corresponding author. Medical Equipment Marketing Department JEOL Ltd., 2-11-1 Inadaira, Musashimurayama, Tokyo, 208-0023, Japan.Medical Equipment Business Operations, JEOL Ltd., Tokyo, JapanMedical Equipment Business Operations, JEOL Ltd., Tokyo, JapanMedical Equipment Business Operations, JEOL Ltd., Tokyo, JapanFaculty of Pharmaceutical Sciences, Tokyo University of Science, Chiba, JapanLaboratories Division, Shimane University Hospital, Shimane, Japan2-Methoxyestradiol (2ME) is involved in the pathogenesis of preeclampsia and antitumor activity. In addition to its low concentration in healthy human serum, presence of isomers makes quantification of 2ME for clinical research and laboratory medicine difficult. The objective of this study was to develop a highly sensitive and accurate method for quantifying 2ME using LC-MS/MS combined with derivatization with 1-(2,4-dinitro-5-fluorophenyl)-4,4-dimethylpiperazinium iodide (MPDNP-F). This approach significantly enhanced the detectability of 2ME in positive electrospray ionization-tandem mass spectrometry (ESI-MS/MS) and enabled the chromatographic separation of 2ME from isomeric metabolites possessing a methoxy group, including 4-methoxyestradiol, 3-O-methyl 2-hydroxyestradiol, and 3-O-methyl 4-hydroxyestradiol (3M4OH). Although the derivatized 2ME and 3M4OH were closely eluted under the optimized LC conditions, the different fragmentation patterns of these isomers during MS/MS allowed their distinction. The lower limit of quantification for 2ME was 2.5 pg/mL, indicating a satisfactory sensitivity. These findings demonstrated that this LC-MS/MS method combined with the MPDNP-F derivatization can provide accurate and highly sensitive quantification of 2ME.http://www.sciencedirect.com/science/article/pii/S2352551724000933LC-MS/MS2-MethoxyestradiolMPDNP-FEstrogenderivatization |
| spellingShingle | Koji Takahashi Masaki Takiwaki Seketsu Fukuzawa Yoshikuni Kikutani Kentaro Abe Tatsuya Higashi Hironori Kobayashi Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometry Practical Laboratory Medicine LC-MS/MS 2-Methoxyestradiol MPDNP-F Estrogen derivatization |
| title | Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometry |
| title_full | Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometry |
| title_fullStr | Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometry |
| title_full_unstemmed | Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometry |
| title_short | Development of an accurate and sensitive assay for 2-methoxyestradiol using derivatization and liquid chromatography-tandem mass spectrometry |
| title_sort | development of an accurate and sensitive assay for 2 methoxyestradiol using derivatization and liquid chromatography tandem mass spectrometry |
| topic | LC-MS/MS 2-Methoxyestradiol MPDNP-F Estrogen derivatization |
| url | http://www.sciencedirect.com/science/article/pii/S2352551724000933 |
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