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

	Peaks	t_R (min)	Precursor Ion	Product Ion	Product Ion
1.	Creatinine	0.537	114.0	44.3	86.0
2.	Creatine	1.232	132.1	43.3	90.2

Column

Raptor 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

5 mM Ammonium formate in 20:80 water:acetonitrile

Time (min)	Flow (mL/min)	%A
0.00	0.5	100
1.5	Stop

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC
Sample Preparation	Endogenous 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.

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