1. Academic Validation
  2. Simultaneous analysis of regorafenib and sorafenib and three of their metabolites in human plasma using LC-MS/MS

Simultaneous analysis of regorafenib and sorafenib and three of their metabolites in human plasma using LC-MS/MS

  • J Pharm Biomed Anal. 2017 Aug 5;142:42-48. doi: 10.1016/j.jpba.2017.04.053.
Marie Allard 1 Nihel Khoudour 1 Benoît Rousseau 2 Charlotte Joly 2 Charlotte Costentin 3 Benoît Blanchet 4 Christophe Tournigand 2 Anne Hulin 5
Affiliations

Affiliations

  • 1 Laboratory of Pharmacology, AP-HP, GH H Mondor, Université Paris-Est Créteil, Créteil, France.
  • 2 Oncology Department, AP-HP, GH H Mondor, Université Paris-Est Créteil, Créteil, France.
  • 3 Hepatology Department, AP-HP, GH H Mondor, Université Paris-Est Créteil, Créteil, France.
  • 4 Laboratory of Pharmacology, AP-HP, GH Cochin, Université Paris Descartes, France.
  • 5 Laboratory of Pharmacology, AP-HP, GH H Mondor, Université Paris-Est Créteil, Créteil, France. Electronic address: anne.hulin@aphp.fr.
Abstract

A new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, performed by electrospray ionization in positive mode using a triple quadrupole mass spectrometry, has been developed and validated for the simultaneous determination of regorafenib (REGO), its two metabolites regorafenib-M2 and regorafenib-M5, sorafenib (SORA), and its N-oxide metabolite in human plasma. Separation is achieved on an Hypersil Gold® column using a gradient elution of 10mM ammonium formate containing 0.1% formic acid (A) and acetonitrile containing 0.1% formic acid (B) at a flow rate of 0.3mL/min. After addition of two internal standards and a protein precipitation, the supernatant is diluted two-fold in a 0.1% (v/v) formic acid solution. Two selected reaction monitoring transitions are used, for each analyte, one for quantitation and the second one for confirmation. The standard curves are ranged from 50 to 5 000ng/mL for REGO and its metabolites and 80 to 5 000ng/mL for SORA and its metabolite and were fitted to a 1/x weighted linear regression model. The method also showed satisfactory results in terms of sensitivity, specificity, precision (intra- and inter-day CV from 2.4 to 10.2%), accuracy (from 91.0 to 111.7%), recovery as well as stability of the analytes under various conditions. The method is usually used in clinical practice in order to improve the SORA treatment for renal carcinoma, REGO treatment for colorectal Cancer and both for hepatocellular carcinoma.

Keywords

LC–MS/MS; Regorafenib; Regorafenib-M2; Regorafenib-M5; Sorafenib; Sorafenib-N-oxide.

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