Bile Acids with Isomer Separation in Human Plasma on Raptor C18 (1.8 μm) by LC-MS/MS

Column

Raptor C18 (cat.# 9304252)

Dimensions:

50 mm x 2.1 mm ID

Particle Size:

1.8 µm

Pore Size:

90 Å

Guard Column:

UltraShield UHPLC precolumn filter, 0.2 µm (cat.# 25810)

Temp.:

60 °C

Standard/Sample

Diluent:

70:30 Water:methanol

Inj. Vol.:

3 µL

Mobile Phase

5 mM Ammonium acetate in water

50:50 Acetonitrile:methanol

Time (min)	Flow (mL/min)	%A	%B
0.00	0.5	65	35
2.00	0.5	60	40
2.50	0.5	55	45
3.50	0.5	50	50
4.60	0.5	45	55
5.70	0.5	20	80
5.90	0.8*	5	95
6.50	0.8*	5	95
6.51	0.5	65	35
8.50	0.5	65	35

Detector	MS/MS
Ion Mode:	ESI-
Mode:	MRM
Instrument	UHPLC
Notes	*The flow rate was increased to 0.8 mL/min to more thoroughly flush phospholipids from the analytical column, thereby reducing matrix effects.

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