Determination of tamoxifen and endoxifen in dried blood spots using LC-MS/MS and the effect of coated DBS cards on recovery and matrix effects.

BACKGROUND: We developed an HPLC-MS/MS method to quantify tamoxifen (2.5-250 ng/ml) and its metabolite (Z)-endoxifen (0.5-50 ng/ml) in dried blood spots. RESULTS: Extraction recovery of both analytes from Whatman DMPK-A cards was 100% and consistent over time, however, recovery of (Z)-endoxifen from Whatman 903 cards was incomplete and increased upon storage. When SDS, a constituent of the DMPK-A coating, was present during the extraction, recovery improved. The method using DMPK-A cards was validated using bioanalytical guidelines. Additionally, influence of haematocrit (0.29-0.48 L/L), spot volume (20-50 µl) and homogeneity was within limits and both analytes were stable in DBS for at least 4 months. CONCLUSIONS: The method for the quantification of tamoxifen and (Z)-endoxifen in DBS collected on DMPK-A cards was successfully validated.

Identification of drug metabolites in human plasma or serum integrating metabolite prediction, LC-HRMS and untargeted data processing.

BACKGROUND: Comprehensive identification of human drug metabolites in first-in-man studies is crucial to avoid delays in later stages of drug development. We developed an efficient workflow for systematic identification of human metabolites in plasma or serum that combines metabolite prediction, high-resolution accurate mass LC-MS and MS vendor independent data processing. Retrospective evaluation of predictions for 14 (14)C-ADME studies published in the period 2007-January 2012 indicates that on average 90% of the major metabolites in human plasma can be identified by searching for accurate masses of predicted metabolites. Furthermore, the workflow can identify unexpected metabolites in the same processing run, by differential analysis of samples of drug-dosed subjects and (placebo-dosed, pre-dose or otherwise blank) control samples. To demonstrate the utility of the workflow we applied it to identify tamoxifen metabolites in serum of a breast cancer patient treated with tamoxifen. RESULTS & CONCLUSION: Previously published metabolites were confirmed in this study and additional metabolites were identified, two of which are discussed to illustrate the advantages of the workflow.