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Sample-multiplexing by derivatization using multiple analogous reagents for enhancing throughput in LC/ESI-MS/MS assay of steroids: Plasma 17α-hydroxyprogesterone as an example.
Kamemura, Miki; Yokota, Mai; Ogawa, Shoujiro; Sugiura, Takahiro; Higashi, Tatsuya.
Afiliação
  • Kamemura M; Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan.
  • Yokota M; Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan.
  • Ogawa S; Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan.
  • Sugiura T; Toyohashi Municipal Hospital, 50 Hachiken Nishi, Aotake-Cho, Toyohashi-shi, Aichi 441-8570, Japan.
  • Higashi T; Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan. Electronic address: higashi@rs.tus.ac.jp.
Article em En | MEDLINE | ID: mdl-32335501
ABSTRACT
The measurements of steroids in biological fluids are of importance for the diagnosis and treatment of many diseases. Liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) has a high specificity and accuracy for the steroid analysis, whereas it has a lower analysis throughput, which could become a big issue in clinical practice. One of the promising solutions to this issue is the multiplexing of samples in the same injection. In this study, the utility of the sample-multiplexing by the derivatization using multiple analogous reagents was evaluated for enhancing the throughput of the LC/ESI-MS/MS assays of steroids. The plasma 17α-hydroxyprogesterone (17OHP), which is a diagnostic marker for the 21-hydroxylase deficiency, was chosen as the model analyte. The four plasma samples (20 µL each) were separately derivatized with one of four different analogous Girard-type reagents, combined, then injected together into the LC/ESI-MS/MS. By this method, four plasma samples could be analyzed within a single LC run. The developed method could significantly reduce the total LC run time (about 2/5 for 32 samples, compared with the conventional method) with a satisfactory sensitivity (lower limit of quantification 0.5 ng/mL), precision (intra- and inter-assay RSDs ≤ 4.0% and ≤ 3.5%, respectively) and accuracy (97.6-106.7%), and negligible matrix effect. The developed method had a satisfactory applicability for the quantification of 17OHP in the cord plasma samples.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Hiperplasia Suprarrenal Congênita / 17-alfa-Hidroxiprogesterona / Espectrometria de Massas em Tandem / Ensaios de Triagem em Larga Escala Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Humans Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Japão País de publicação: HOLANDA / HOLLAND / NETHERLANDS / NL / PAISES BAJOS / THE NETHERLANDS

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Hiperplasia Suprarrenal Congênita / 17-alfa-Hidroxiprogesterona / Espectrometria de Massas em Tandem / Ensaios de Triagem em Larga Escala Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Humans Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Japão País de publicação: HOLANDA / HOLLAND / NETHERLANDS / NL / PAISES BAJOS / THE NETHERLANDS