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Creatine and Creatinine in Human Plasma on Raptor HILIC-Si by LC-MS/MS

LC_CF0722
PeakstR (min)Precursor IonProduct IonProduct Ion
1.Creatinine0.537114.044.386.0
2.Creatine1.232132.143.390.2
ColumnRaptor HILIC-Si (cat.# 9310A52)
Dimensions:50 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:UltraShield UHPLC precolumn filter 0.2 µm (cat.# 25810)
Temp.:40 °C
Standard/Sample
Diluent:20:80 Water:acetonitrile
Conc.:Endogenous levels
Inj. Vol.:0.2 µL
Mobile Phase
A:5 mM Ammonium formate in 20:80 water:acetonitrile
Time (min)Flow (mL/min)%A
0.000.5100
1.5Stop
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentUHPLC
Sample PreparationEndogenous levels of creatinine and creatine in human plasma were determined using a single protein precipitation step followed by LC-MS/MS analysis. A 50 μL aliquot of human plasma (K2EDTA) was mixed with 950 μL acetonitrile. After vortexing and centrifuging at 4300 rpm for 10 min, 200 μL of the supernatant was transferred to a new vial and mixed with 50 μL of water. Centrifugation was performed again before injection.