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Determination of Rac inhibitor Ehop‐016 in mouse plasma by ultra‐performance liquid chromatography tandem mass spectrometry (987.2)
Author(s) -
HumphriesBickley Tessa,
Corujo Francheska,
CastilloPichardo Linette,
HernandezO'Farrill Eliud,
Vlaar Cornelis,
RodriguezOrengo Jose,
Dharmawardhane Suranganie
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.987.2
Subject(s) - chemistry , chromatography , formic acid , in vivo , tandem mass spectrometry , high performance liquid chromatography , electrospray ionization , selected reaction monitoring , mass spectrometry , pharmacokinetics , pharmacology , microbiology and biotechnology , biology , medicine
The Rho GTPase Rac is an important regulator of cancer cell migration and cell invasion; processes required for metastatic progression. We previously characterized the small molecule Ehop‐016 as a novel Rac inhibitor in metastatic breast cancer cells in‐vitro (Montalvo‐Ortiz, et al., 2012). To investigate the efficacy of EHop‐016 in‐vivo, we used a mouse model of experimental metastasis, where mice with MDA‐MB‐435 mammary fat pad tumors were treated with intraperitoneal EHop‐016. At 25 and 40 mg/kg BW, EHop‐016 significantly inhibited mammary tumor growth and metastasis. In order to characterize the pharmacokinetics of Ehop‐016, we developed a rapid and sensitive method for the quantitation of Ehop‐016 in mouse plasma by ultra high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC/MS/MS). Plasma samples were pretreated with acetonitrile (ACN) to extract matrix proteins prior to injection into UPLC/MS/MS. Separation was carried out on an Agilent Poroshell 120 EC‐C18 column (3.0 x 50) mm using a mobile phase of 50% ACN/50% methanol/0.1% formic acid and 1mM ammonium fluoride. Ehop‐016 was identified from its accurate mass and retention times from the acquired full‐scan chromatogram and quantified by its peak areas. The linear range for the determination of analytes was 5 ‐ 1000 ng/mL. Ehop‐016 was quantified in samples from the in‐vivo study using this method.