Simultaneous determination of eight protein anabolic hormones in human plasma by ultrafast liquid chromatography- tandem mass spectrometry

AIM To develop a sensitive and accurate method for the simultaneous determination of 8 protein anabolic hormones in human plasma by ultrafast liquid chromatography- tandem mass spectrometry(UFLC-MS/MS)with isotope internal standard dilution technique and solid-phase extraction.METHODS Proteins in plasma sample were precipitated by methanol and cleaned by Waters Oasis~(?) HLB solid-phase extraction cartridge.The separation was performed on a Shim-pack XR-ODSⅡcolumn(75 mm×2.0 mm,2.2μm) with a linear gradient elution program using acetonitrile (containing 0.1%formic acid) and 0.1%formic acid aqueous solution as mobile phase.Electrospray ionization was applied and operated in the positive ion multiple reaction monitoring(MRM) mode.The quantitation was carried on by methyltestosterone-D_3 isotope internal standard dilution technique.RESULTS The calibration curves were in good linearity for the 8 analytes in the each linearity range with the correlative coefficients more than 0.999.Limits of quantitation (LOQs,R_(S,N)≥10) were in the range of 0.5 -1.0μg·L~(-1),and the relative recoveries were between 95.0%and 105.0%.The intra-day relative standard deviations(RSDs) for assaying the plasma sample containing three concentrations of the 8 analytes were all lower than 6.0%,and inter-day RSDs were all lower than 10.0%.CONCLUSION The developed method is specific,sensitive and simple,and is conformed to the requirements of biological specimen analyses.