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Validation and application of a liquid chromatography–tandem mass spectrometric method for the determination of GDC‐0834 and its metabolite in human plasma using semi‐automated 96‐well protein precipitation
Author(s) -
Shin Young G.,
Jones Steve A.,
Murakami Stan C.,
Liu Lichuan,
Wong Harvey,
Buonarati Michael H.,
Hop Cornelis E. C. A.
Publication year - 2012
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.2716
Subject(s) - metabolite , chromatography , chemistry , protein precipitation , liquid chromatography–mass spectrometry , active metabolite , tandem mass spectrometry , mass spectrometry , detection limit , pharmacokinetics , pharmacology , biochemistry , medicine
A liquid chromatographic–tandem mass spectrometric (LC‐MS/MS) method was developed and validated for the determination of GDC‐0834 and its amide hydrolysis metabolite (M1) in human plasma to support clinical development. The method consisted of semi‐automated 96‐well protein precipitation extraction for sample preparation and LC‐MS/MS analysis in positive ion mode using TurboIonSpray® for analysis. D6 ‐GDC‐0834 and D6 ‐M1 metabolite were used as internal standards. A linear regression (weighted 1/concentration 2 ) was used to fit calibration curves over the concentration range of 1 – 500 ng/mL for both GDC‐0834 and M1 metabolite. The accuracy (percentage bias) at the lower limit of quantitation (LLOQ) was 5.20 and 0.100% for GDC‐0834 and M1 metabolite, respectively. The precision (CV) for samples at the LLOQ was 3.13–8.84 and 5.20–8.93% for GDC‐0834 and M1 metabolite, respectively. For quality control samples at 3, 200 and 400 ng/mL, the between‐run CV was ≤7.38% for GDC‐0834 and ≤8.20% for M1 metabolite. Between run percentage bias ranged from −2.76 to 6.98% for GDC‐0834 and from −6.73 to 2.21% for M1 metabolite. GDC‐0834 and M1 metabolite were stable in human plasma for 31 days at −20 and −70°C. This method was successfully applied to support a GDC‐0834 human pharmacokinetic‐based study. Copyright © 2012 John Wiley & Sons, Ltd.