LC/MS/MS of Digoxin and Digitoxin in Serum or Plasma on Titan™ C18 after SPE with HybridSPE®-Phospholipid, Minimization of Matrix Effects

LC/MS/MS of Digoxin and Digitoxin in Serum or Plasma on Titan™ C18 after SPE with HybridSPE®-Phospholipid, Minimization of Matrix Effects

Conditions

sample preparation SPE (Solid Phase Extraction)
sample/matrix rat plasma or human serum spiked at 0.05 ng/mL with digitoxin and digoxin
SPE well plate HybridSPE-PLus 96-well plate, 50 mg/well (575659-U)
sample addition 100 µL spiked rat plasma followed by 300 µL 1% formic acid in acetonitrile
elution apply vacuum at 10 in Hg for 4 min
column Titan C18, 10 cm x 2.1 mm I.D., 1.9 µm particles (577124-U)
mobile phase 10 mM ammonium formate in methanol:water (80:20)
flow rate 0.2 mL/min
pressure 4550 psi (314 bar)
injection 2 µL
detector MS, ESI(+), MRM 798.5/651.5 (digoxin), 782.5/635.5 or 243.2 (digtoxin)

Description

Analysis Note The cardiac glycosides digitoxin and digoxin are widely prescribed for treating congestive heart failure. Immunoassay-based methods are not able to effectively differentiate these two compounds due to their similar chemical structures. This application demonstrates a rapid, selective and sensitive LC/MS/MS method for determination of digitoxin and digoxin in plasma with a simple sample cleanup technique using HybridSPE-PLus for siµLtaneous removal of proteins and phospholipids and a Titan C18 column for rapid and efficient UHPLC separation. The highest grade LC-MS solvents were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant CRMs provided reliable quantification.
Categories Analytical Chromatography, SPE Applications, Cardiac drugs
Featured Industry Clinical
Life Science and Biopharma
Legal Information HybridSPE is a registered trademark of Sigma-Aldrich Co. LLC, Titan is a trademark of Sigma-Aldrich Co. LLC
suitability application for LC-MS, application for SPE

Materials

     
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