{"title":"利用超高效液相色谱-质谱 3测定法同时测定甲氨蝶呤和 7-羟基甲氨蝶呤，并采用多级碎片化技术提高灵敏度","authors":"Huaqiang Li, Xujian Duan, Feifei Wu, Lei Yin","doi":"10.1002/jssc.70020","DOIUrl":null,"url":null,"abstract":"<div>\n \n <p>A selective ultra-high performance liquid chromatography tandem mass spectrometry cubed (UHPLC/MS<sup>3</sup>) assay for simultaneous determination of methotrexate (MTX) and 7-hydroxy-methotrexate (7-OH-MTX) in human plasma was developed and validated. After protein precipitation with methanol, chromatographic separation of MTX, MTX-d<sub>3</sub>, and 7-OH-MTX was performed on a Waters AcQuity UPLC-BEH column (2.1 × 50 mm I.D., 1.7 µm) with gradient elution. MRM<sup>3</sup> transition was used for the detection of MTX (<i>m/z</i> 455.2→308.0→175.1) and 7-OH-MTX (<i>m/z</i> 471.3→191.0→148.1). The linear range of UHPLC/MS<sup>3</sup> assay for the determination of MTX and 7-OH-MTX was 0.5–300 ng/mL (<i>R</i><sup>2</sup> ≥ 0.99) and 5–1500 ng/mL (<i>R</i><sup>2</sup> ≥ 0.99), respectively. The enhanced selectivity and sensitivity are the novelties of the developed UHPLC/MS<sup>3</sup> assay. The analytical method was successfully applied to simultaneous determination of MTX and its major metabolite 7-OH-MTX in real human plasma samples.</p>\n </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"47 21","pages":""},"PeriodicalIF":2.8000,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Simultaneous Determination of Methotrexate and 7-Hydroxy-Methotrexate by UHPLC-MS3 Assay Coupled With Multiple Stage Fragmentation to Enhance Sensitivity\",\"authors\":\"Huaqiang Li, Xujian Duan, Feifei Wu, Lei Yin\",\"doi\":\"10.1002/jssc.70020\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n <p>A selective ultra-high performance liquid chromatography tandem mass spectrometry cubed (UHPLC/MS<sup>3</sup>) assay for simultaneous determination of methotrexate (MTX) and 7-hydroxy-methotrexate (7-OH-MTX) in human plasma was developed and validated. After protein precipitation with methanol, chromatographic separation of MTX, MTX-d<sub>3</sub>, and 7-OH-MTX was performed on a Waters AcQuity UPLC-BEH column (2.1 × 50 mm I.D., 1.7 µm) with gradient elution. MRM<sup>3</sup> transition was used for the detection of MTX (<i>m/z</i> 455.2→308.0→175.1) and 7-OH-MTX (<i>m/z</i> 471.3→191.0→148.1). The linear range of UHPLC/MS<sup>3</sup> assay for the determination of MTX and 7-OH-MTX was 0.5–300 ng/mL (<i>R</i><sup>2</sup> ≥ 0.99) and 5–1500 ng/mL (<i>R</i><sup>2</sup> ≥ 0.99), respectively. The enhanced selectivity and sensitivity are the novelties of the developed UHPLC/MS<sup>3</sup> assay. The analytical method was successfully applied to simultaneous determination of MTX and its major metabolite 7-OH-MTX in real human plasma samples.</p>\\n </div>\",\"PeriodicalId\":17098,\"journal\":{\"name\":\"Journal of separation science\",\"volume\":\"47 21\",\"pages\":\"\"},\"PeriodicalIF\":2.8000,\"publicationDate\":\"2024-11-08\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of separation science\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/jssc.70020\",\"RegionNum\":3,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of separation science","FirstCategoryId":"5","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/jssc.70020","RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
Simultaneous Determination of Methotrexate and 7-Hydroxy-Methotrexate by UHPLC-MS3 Assay Coupled With Multiple Stage Fragmentation to Enhance Sensitivity
A selective ultra-high performance liquid chromatography tandem mass spectrometry cubed (UHPLC/MS3) assay for simultaneous determination of methotrexate (MTX) and 7-hydroxy-methotrexate (7-OH-MTX) in human plasma was developed and validated. After protein precipitation with methanol, chromatographic separation of MTX, MTX-d3, and 7-OH-MTX was performed on a Waters AcQuity UPLC-BEH column (2.1 × 50 mm I.D., 1.7 µm) with gradient elution. MRM3 transition was used for the detection of MTX (m/z 455.2→308.0→175.1) and 7-OH-MTX (m/z 471.3→191.0→148.1). The linear range of UHPLC/MS3 assay for the determination of MTX and 7-OH-MTX was 0.5–300 ng/mL (R2 ≥ 0.99) and 5–1500 ng/mL (R2 ≥ 0.99), respectively. The enhanced selectivity and sensitivity are the novelties of the developed UHPLC/MS3 assay. The analytical method was successfully applied to simultaneous determination of MTX and its major metabolite 7-OH-MTX in real human plasma samples.
期刊介绍：
The Journal of Separation Science (JSS) is the most comprehensive source in separation science, since it covers all areas of chromatographic and electrophoretic separation methods in theory and practice, both in the analytical and in the preparative mode, solid phase extraction, sample preparation, and related techniques. Manuscripts on methodological or instrumental developments, including detection aspects, in particular mass spectrometry, as well as on innovative applications will also be published. Manuscripts on hyphenation, automation, and miniaturization are particularly welcome. Pre- and post-separation facets of a total analysis may be covered as well as the underlying logic of the development or application of a method.
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