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- W2321111963 abstract "A liquid-liquid extraction and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) method to determine thyronines (TH) and thyronamines (TAM) from cell culture media was developed and applied to cell homogenate extraction. The method presented here for extraction of DMEM/F12 cell culture media is a fundamental procedure for a precise determination of 9 TH and 6 TAM from a single LC run. Analytes and internal standards (IS) were extracted from DMEM/F12 w/o phenol-red and IS were extracted from MDCK-1 (Madin-Darby canine kidney) cell homogenates by liquid-liquid extraction using isopropanol-TBME (30:70 v/v). Measurement of TH and TAM was performed during a 10 min run-time using 13C6T4, 13C6T3, 13C6rT3, 13C63,3'T2, and 2H4T1AM as IS. Calibration curves covered 11 calibrators measured as triplicates each for the analysis of the 9 TH, 6 TAM metabolites and the 5 IS were linear and reproducible in the range of 0.12 – 120 nM (R2: 0.991 – 0.999) for all calibrators from cell media. The lower limit of quantification was 0.078 – 0.234 nM. Method validation and robustness for cell media extraction were demonstrated by analysis of precision, accuracy, process efficiency, matrix effects, recoveries as well as intra- and interassay stability. Application of the validated extraction method to cell homogenates resulted in solid recoveries > 80% for all IS and no substantial matrix effects. This validated, sensitive and interaction-free LC-MS/MS method allows rapid analysis and accurate determination of TH and TAM from DMEM/F12 w/o phenol red conditioned media and seems to be easily transferable to cell culture homogenates." @default.
- W2321111963 created "2016-06-24" @default.
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- W2321111963 date "2015-03-12" @default.
- W2321111963 modified "2023-10-18" @default.
- W2321111963 title "Quantitative analysis of thyroid hormone metabolites in cell culture samples using LC-MS/MS" @default.
- W2321111963 doi "https://doi.org/10.1055/s-0035-1547742" @default.
- W2321111963 hasPublicationYear "2015" @default.
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