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http://dx.doi.org/10.5806/AST.2007.20.5.424

Determination of thyroid hormones in plasma samples by high performance liquid chromatograph/diode array detector/electrospray ionization mass spectrometer  

Kwak, Sun Young (Process Development Lab., Choongwae Pharma Corp.)
Moon, Myeong Hee (Department of Chemistry, Yonsei University)
Pyo, Heesoo (Bioanalysis & Biotransformation Research Center, Korea Institute Science & Technology)
Publication Information
Analytical Science and Technology / v.20, no.5, 2007 , pp. 424-433 More about this Journal
Abstract
An analytical method for the determination of thyroid hormones in plasma samples has been studied by solid-phase extraction and high-performance liquid chromatography/diode array detector (DAD)/electrospray ionization (ESI)-mass spectrometer. Seven thyroid hormones were successfully separated by gradient elution on the reverse phase Hypersil ODS column (4.6 mm I.D., 250 mm length, particle size $5{\mu}m$) with ammonium formate buffer and acetonitrile. In addition, these compounds were confirmed by UV spectra and ESI-mass Spectra. The extraction recoveries of thyroid hormones in the plasma sample (at pH 3) were in the range of 74.5-115.7 % with solid-phase extraction by C18, followed by elution with 4 mL of methanol. The calibration curves showed good linearity with the correlation coefficients ($r^2$) varying from 0.9939 to 0.9978 and the detection limits of all analytes were obtained in the range of 20-50 ng/mL (38.1-162.8 pmol/mL). As a result, thyroxine was found in the range of 50.98-112.97 ng/mL in normal plasma samples.
Keywords
thyroid hormones; HPLC/DAD/ESI-MS; SPE; plasma;
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