Solid-phase extraction and high-performance liquid chromatographic determination of tamoxifen and its major metabolites in plasma

MacCallum, J.; Cummings, J.; Dixon, J.M.; Miller, W.R.

Abstract

Tamoxifen (TAM) is a triphenylethylene anti-oestrogen, commonly used in the treatment of breast cancer. Patients receiving tamoxifen therapy may experience both de novo and acquired resistance. As one of the mechanisms for this may be extensive peripheral bio-transformation of tamoxifen, there has been considerable interest in the pharmacokinetics and metabolism of tamoxifen. A reversed-phase high-performance liquid chromatography separation has been developed to determine the levels of tamoxifen and its major metabolites in human plasma. The method is highly sensitive (2 ng/ml) and selective for tamoxifen, cis-tamoxifen (CIS), 4-hydroxytamoxifen (4-OH) and desmethyltamoxifen (DMT). A μBondapak C18 10 μm column (30 cm × 3.9 mm I.D.) was used, with a mobile phase of methanol-1% triethylamine at pH 8 (89:11, v/v). Sample preparation was carried out using a C2 (500 mg sorbent, 3 ml reservoirs) solid phase extraction method, and extraction efficiencies were approximately 60% for TAM and its metabolites. Accuracy and precision, as determined by spiking plasma samples with a mixture of tamoxifen and its metabolites, ranged from 85–110% (± 5–10%) at 1 μg/ml, 101–118% (± 8–20%) at 0.1 μg/ml and 111–168% (± 43–63%) at 0.01 μg/ml. Results from 59 patients show mean values of 54 ng/ml for 4-OH; 190 ng/ml for DMT; 93 ng/ml for TAM and 30 ng/ml for CIS (detected in three patients only). This methodology can be applied routinely to the determination of TAM and its metabolites in plasma from patients undergoing therapy.

Citation

MacCallum, J., Cummings, J., Dixon, J., & Miller, W. (1996). Solid-phase extraction and high-performance liquid chromatographic determination of tamoxifen and its major metabolites in plasma. Journal of Chromatography B, 678(2), 317-323. https://doi.org/10.1016/0378-4347%2895%2900530-7

• Journal Article Type: Article
• Publication Date: 1996-04
• Deposit Date: May 30, 2022
• Journal: Journal of Chromatography B: Biomedical Sciences and Applications
• Print ISSN: 1570-0232
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 678
• Issue: 2
• Pages: 317-323
• DOI: https://doi.org/10.1016/0378-4347%2895%2900530-7
• Public URL: http://researchrepository.napier.ac.uk/Output/2874947