SIMULTANEOUS QUANTIFICATION OF TAMOXIFEN AND 4-HYDROXY-N-DESMETHYLTAMOXIFEN LEVELS IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY: DEVELOPMENT AND APPLICATION IN BREAST CANCER PATIENTS

  • Yahdiana Harahap Bioavailability/Bioequivalence Laboratory, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.
  • Nathania Leony Bioavailability/Bioequivalence Laboratory, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.
  • Letare Merry Chresia Silalahi Bioavailability/Bioequivalence Laboratory, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.
  • Rizka Andalusia Pharmacy Installation, Dharmais Cancer Hospital, Jakarta, 11420, Indonesia.

Abstract

Objective: This study was aimed to develop a highly sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method
for the simultaneous quantification of tamoxifen, endoxifen, and clomiphene (internal standard) levels in human plasma.
Methods: Plasma samples from 40 patients with breast cancer were prepared through liquid–liquid extraction in ethyl acetate, and chromatographic
separation was performed on an Acquity UPLC BEH C18 column (1.7 μm, 2.1 mm×100 mm). The sample was eluted within 6.50 min at a flow rate of
0.200 ml/min under a gradient of 0.2% formic acid and acetonitrile. The analytes were then quantified through triple quadrupole MS with electrospray
ionization in the positive ion mode and multiple reaction monitoring.
Results: The method was successfully used to quantify the plasma levels of tamoxifen and endoxifen, yielding respective measurement ranges
of 42.19–249.23 ng/ml and 1.52–26.62 ng/ml. Overall, this detection method was sensitive, with lower limits of quantitation of 0.625 ng/ml for
tamoxifen and 0.125 ng/ml for endoxifen. In addition, this method was precise and accurate, with a between-run variation range of 5.19–12.38% and
%diff range of −10.82%–+13.10%.
Conclusion: This method demonstrated that approximately 80% of patients had plasma levels of endoxifen exceeding the expected clinical threshold
(5.9 ng/ml). This rapid and simple method could be used to monitor tamoxifen metabolism and adjust doses as needed to achieve maximum
effectiveness.

Keywords: 4-Hydroxy-N-desmethyltamoxifen (endoxifen), Breast cancer, Clomiphene, Liquid chromatography-tandem mass spectrometry, Tamoxifen, Validation.

References

1. Hu R, Hilakivi-Clarke L, Clarke R. Molecular mechanisms of
tamoxifen-associated endometrial cancer (Review). Oncol Lett
2015;9:1495-501.
2. Tchu SM, Lynch KL, Wu AH. Therapeutic drug monitoring of
tamoxifen using LC-MS/MS. Methods Mol Biol 2012;902:211-22.
3. Goodman L, Gilman A, Brunton L, Chabner B, Knollman B. Goodman
and Gilman’s the Pharmacological Basis of Therapeutics. 12th.
New York: McGraw-Hill; 2012.
4. Antunes MV, Raymundo S, de Oliveira V, Staudt DE, Gössling G,
Peteffi GP, et al. Ultra-high performance liquid chromatography
tandem mass spectrometric method for the determination of tamoxifen,
N-desmethyltamoxifen, 4-hydroxytamoxifen and endoxifen in dried
blood spots-development, validation and clinical application during
breast cancer adjuvant therapy. Talanta 2015;132:775-84.
5. Arellano C, Allal B, Goubaa A, Roché H, Chatelut E. An UPLC-MS/
MS method for separation and accurate quantification of tamoxifen and
its metabolites isomers. J Pharm Biomed Anal 2014;100:254-61.
6. Tré-Hardy M, Capron A, Antunes MV, Linden R, Wallemacq P. Fast
method for simultaneous quantification of tamoxifen and metabolites
in dried blood spots using an entry level LC-MS/MS system. Clin
Biochem 2016;49:1295-8.
7. Antunes MV, Rosa DD, Viana Tdos S, Andreolla H, Fontanive TO,
Linden R, et al. Sensitive HPLC-PDA determination of tamoxifen
and its metabolites N-desmethyltamoxifen, 4-hydroxytamoxifen and
endoxifen in human plasma. J Pharm Biomed Anal 2013;76:13-20.
8. ter Heine R, Binkhorst L, de Graan AJ, de Bruijn P, Beijnen JH,
Mathijssen RH, et al. Population pharmacokinetic modelling to assess
the impact of CYP2D6 and CYP3A metabolic phenotypes on the
pharmacokinetics of tamoxifen and endoxifen. Br J Clin Pharmacol
2014;78:572-86.
9. Jordan VC. New insights into the metabolism of tamoxifen and
its role in the treatment and prevention of breast cancer. Steroids
2007;72:829-42.
10. Hawse JR, Subramaniam M, Cicek M, Wu X, Gingery A, Grygo SB,
et al. Endoxifen’s molecular mechanisms of action are concentration
dependent and different than that of other anti-estrogens. PLoS One
2013;8:e54613.
11. Binkhorst L, Mathijssen RH, Ghobadi Moghaddam-Helmantel IM, de
Bruijn P, van Gelder T, Wiemer EA, et al. Quantification of tamoxifen
and three of its phase-I metabolites in human plasma by liquid
chromatography/triple-quadrupole mass spectrometry. J Pharm Biomed
Anal 2011;56:1016-23.
12. Binkhorst L, Mathijssen RH, Jager A, van Gelder T. Individualization
of tamoxifen therapy: Much more than just CYP2D6 genotyping.
Cancer Treat Rev 2015;41:289-99.
13. Gjerde J, Kisanga ER, Hauglid M, Holm PI, Mellgren G, Lien EA, et al.
Identification and quantification of tamoxifen and four metabolites
in serum by liquid chromatography-tandem mass spectrometry.
J Chromatogr A 2005;1082:6-14.
14. Dahmane E, Mercier T, Zanolari B, Cruchon S, Guignard N, Buclin T,
et al. An ultra performance liquid chromatography-tandem MS assay
for tamoxifen metabolites profiling in plasma: First evidence of
4’-hydroxylated metabolites in breast cancer patients. J Chromatogr B
Analyt Technol Biomed Life Sci 2010;878:3402-14.
15. Hoskins JM, Carey LA, McLeod HL. CYP2D6 and tamoxifen: DNA
matters in breast cancer. Nat Rev Cancer 2009;9:576-86.
16. Ahmad A, Shahabuddin S, Sheikh S, Kale P, Krishnappa M, Rane RC,
et al. Endoxifen, a new cornerstone of breast cancer therapy:
Demonstration of safety, tolerability, and systemic bioavailability in
healthy human subjects. Clin Pharmacol Ther 2010;88:814-7.
17. Li W, Zhang J, Tse FL. Handbook of LC-MS Bioanalysis: New Jersey:
John Wiley and Sons; 2013.
18. Madlensky L, Natarajan L, Tchu S, Pu M, Mortimer J, Flatt SW, et al.
Tamoxifen metabolite concentrations, CYP2D6 genotype and breast
cancer outcomes. Clin Pharmacol Ther 2011;89:718-25.
19. Qureshi SS, Shah K, Upmanyu N. Prevalence and risk factor of
polycystic ovarian syndrome. Asian J Pharm Clin Res 2016;9:23-5.
20. European Medicines Agency. Guideline on Bioanalytical Method
Validation: London: European Medicines Agency; 2012.
21. Satria D, Hasibuan PA, Sitorus P. Anticancer activity of B-Sitosterol
from Plectranthus amboinicus (Lour.spreng) leaves: In vitro and
in silico studies. Asian J Pharm Clin Res 2017;10:306-8.
22. Pappachen LK, Zachariah SM, Chandran D. Insilico design, synthesis
and characterization of some novel benzothiazole derivatives as anticancer
agents. Asian J Pharm Clin Res 2017;10:150-5.
Statistics
35 Views | 49 Downloads
How to Cite
Harahap, Y., Leony, N., Silalahi, L. M. C., & Andalusia, R. (2018). SIMULTANEOUS QUANTIFICATION OF TAMOXIFEN AND 4-HYDROXY-N-DESMETHYLTAMOXIFEN LEVELS IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY: DEVELOPMENT AND APPLICATION IN BREAST CANCER PATIENTS. International Journal of Applied Pharmaceutics, 10(1), 370-375. https://doi.org/10.22159/ijap.2018.v10s1.82
Section
Original Article(s)