Development and validation of UPLC method for WST-1 cell viability assay and its application to MCTT HCE (TM) eye irritation test for colorful substances
DC Field | Value | Language |
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dc.contributor.author | Joo, Kyung-Mi | - |
dc.contributor.author | Kim, Seolyeong | - |
dc.contributor.author | Koo, Ye Ji | - |
dc.contributor.author | Lee, Miri | - |
dc.contributor.author | Lee, Su-Hyun | - |
dc.contributor.author | Choi, Dalwoong | - |
dc.contributor.author | Lim, Kyung-Min | - |
dc.date.accessioned | 2021-09-01T05:08:12Z | - |
dc.date.available | 2021-09-01T05:08:12Z | - |
dc.date.created | 2021-06-18 | - |
dc.date.issued | 2019-10 | - |
dc.identifier.issn | 0887-2333 | - |
dc.identifier.uri | https://scholar.korea.ac.kr/handle/2021.sw.korea/62752 | - |
dc.description.abstract | WST-1 [Water Soluble Tetrazolium-1; 2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenye-2H-tetrazolium, monosodium salt)] is widely used in the cell viability assays replacing MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). A water-soluble formazan dye (4-[1-(4-Iodophenyl)-5-(4-nitrophenyl)formaz-3-yl]-1,3-benzene disulfonate, disodium salt) is produced from the reduction of WST-1 tetrazolium, of which optical density at 450 nm is measured to evaluate cell viability. Colorful substances may interfere with spectrometric measurement, and a method to specifically detect WST-1 formazan is required. Here, a simple, rapid, sensitive, and specific ultra-performance liquid chromatography coupled to UV detector (UPLC-UV) was developed and validated for the WST-1 formazan. For the application to cell viability assay, the supernatant from WST-1 assay was injected without sample preparation procedure and a single run was completed within 5 min. Chromatographic separation was achieved on BEH C18 column (1.7 mu m, 2.1 x 50 mm) using gradient elution with the mobile phase of water and acetonitrile. The standard curves were linear over the concentration range of 2.5-120 mu g/mL WST-1 formazan, which encompasses WST-1 formazan concentrations from 2% cell viability to 2 fold of 100% cell viability. The intra- and inter-day precisions were measured to be below 5% and accuracies were within the range of 91.8-104.9%. The validated method was successfully applied to the test of colorful substances in vitro eye irritation test with a human cornea-like epithelium, and in vitro cytotoxicity in HaCaT, human keratinocyte cell line. | - |
dc.language | English | - |
dc.language.iso | en | - |
dc.publisher | PERGAMON-ELSEVIER SCIENCE LTD | - |
dc.subject | SULFONATED TETRAZOLIUM SALT | - |
dc.subject | MEMBRANE ELECTRON-TRANSPORT | - |
dc.subject | CORNEA-LIKE EPITHELIUM | - |
dc.subject | COLORIMETRIC ASSAY | - |
dc.subject | SOLUBLE FORMAZAN | - |
dc.subject | MTT | - |
dc.subject | REDUCTION | - |
dc.subject | TOXICITY | - |
dc.subject | MODEL | - |
dc.subject | CLASSIFICATION | - |
dc.title | Development and validation of UPLC method for WST-1 cell viability assay and its application to MCTT HCE (TM) eye irritation test for colorful substances | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Choi, Dalwoong | - |
dc.identifier.doi | 10.1016/j.tiv.2019.06.017 | - |
dc.identifier.scopusid | 2-s2.0-85067956062 | - |
dc.identifier.wosid | 000486105900045 | - |
dc.identifier.bibliographicCitation | TOXICOLOGY IN VITRO, v.60, pp.412 - 419 | - |
dc.relation.isPartOf | TOXICOLOGY IN VITRO | - |
dc.citation.title | TOXICOLOGY IN VITRO | - |
dc.citation.volume | 60 | - |
dc.citation.startPage | 412 | - |
dc.citation.endPage | 419 | - |
dc.type.rims | ART | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Toxicology | - |
dc.relation.journalWebOfScienceCategory | Toxicology | - |
dc.subject.keywordPlus | SULFONATED TETRAZOLIUM SALT | - |
dc.subject.keywordPlus | MEMBRANE ELECTRON-TRANSPORT | - |
dc.subject.keywordPlus | CORNEA-LIKE EPITHELIUM | - |
dc.subject.keywordPlus | COLORIMETRIC ASSAY | - |
dc.subject.keywordPlus | SOLUBLE FORMAZAN | - |
dc.subject.keywordPlus | MTT | - |
dc.subject.keywordPlus | REDUCTION | - |
dc.subject.keywordPlus | TOXICITY | - |
dc.subject.keywordPlus | MODEL | - |
dc.subject.keywordPlus | CLASSIFICATION | - |
dc.subject.keywordAuthor | WST-1 assay | - |
dc.subject.keywordAuthor | UPLC | - |
dc.subject.keywordAuthor | Cytotoxicity assay | - |
dc.subject.keywordAuthor | Method validation | - |
dc.subject.keywordAuthor | Color interference | - |
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