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Supercritical Fluid Chromatography‐Tandem Mass Spectrometry Method Development for the Detection of the Rac/Cdc42 inhibitor MBQ‐167 in Mouse Plasma
Author(s) -
Mar Maldonado Maria,
Bloom Joseph,
HernandezO'Farrill Eliud,
Vlaar Cornelis,
RodriguezOrengo Jose F,
Dharmawardhane Suranganie
Publication year - 2017
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.31.1_supplement.924.3
Subject(s) - chemistry , supercritical fluid chromatography , protein precipitation , chromatography , formic acid , mass spectrometry , electrospray ionization , tandem mass spectrometry , supercritical fluid , high performance liquid chromatography , organic chemistry
Metastasis is the principal cause of death for most cancer patients; however, effective treatment options are limited. Targeting pivotal regulators of metastasis the Rho GTPases Rac and Cdc42 is a rational approach for novel metastasis targeted therapy design. Therefore, we developed the small molecule MBQ‐167 as a promising new Rac/Cdc 42 inhibitor, which inhibits mammary tumor growth and metastasis in immunocompromised mice by ~90% and is ten times more potent than other currently available inhibitors. However, further studies are needed to determine the pharmacokinetic (PK) parameters of MBQ‐167 in a mouse model, which are essential to understand the mechanisms of drug distribution and elimination of this compound. Hence, it is necessary to develop a rapid and sensitive method to quantify MBQ‐167 in plasma. Supercritical fluid chromatography (SFC) uses supercritical carbon dioxide (SCO 2 ) as mobile phase and offers advantages such as high sensitivity, increased resolution, and rapid analysis times. Also, it is an environmentally friendly technique, since is requires less organic solvents compared to High Performance Liquid Chromatography (HPLC). The objective of this study was to develop a method for the quantification of MBQ‐167 in mouse plasma using SFC coupled with electrospray ionization tandem mass spectrometry (SFC/MS/MS). Plasma samples were subjected to protein precipitation with acetonitrile. Separation was performed on an ACQUITY UPC 2TM BEH (3.0 × 100 mm, 1.7 μm) column at 35 °C with a mobile phase consisting of CO 2 and methanol 0.1% formic acid (95:5, v/v) at a flow rate of 1 mL/min. A tandem triple quadrupole mass spectrometer was operated in multiple reaction monitoring (MRM) mode. The method was sensitive with a lower limit of quantification (LLOQ) of 2.5 ng/mL and a lower limit of detection (LOD) of 1.25 ng/mL. Preliminary linearity was observed over the concentration range 2.5–1000 ng/mL. The data obtained supports the use of SFC/MS/MS as a highly specific method to evaluate the pharmacokinetics of MBQ‐167 in mice. Support or Funding Information Susan G Komen for the Cure and Puerto Rico Science, Technology, and Research Trust (PRSTRT) grants (to SD), UPR RCM NIH/NIMHHD grant G12MD007600 (to MM)