GHB and Related Compounds in Human Blood by LC-MS/MS

	Peaks	t_R (min)	Precursor ion	Product ion
1.	γ-Hydroxybutyric acid (GHB)	1.42	105.2	87.0
2.	1,4-Butanediol (1,4-BD)	1.42	91.0	55.0
3.	γ-Butyrolactone (GBL)	1.75	87.0	45.0

Column

Force C18 (cat.# 963431E)

Dimensions:

100 mm x 3.0 mm ID

Particle Size:

3 µm

Pore Size:

100 Å

Guard Column:

Force C18 EXP guard column cartridge 5 mm, 3.0 mm ID, 3 µm (cat.# 963450253)

Temp.:

30 °C

Standard/Sample

Diluent:

Water

Conc.:

500 ng/mL

Inj. Vol.:

10 µL

Mobile Phase

0.5% Formic acid in water

0.5% Formic acid in methanol

Time (min)	Flow (mL/min)	%A	%B
0.00	0.7	95	5
0.50	0.7	95	5
3.00	0.7	50	50
3.01	0.7	95	5
5.00	0.7	95	5

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	HPLC
Sample Preparation	100 μL of whole human blood was fortified at 50 μg/mL with GHB, GBL, 1,4-BD, and GHB-D6 (IS) using 5 μL of 1 mg/mL solutions. The blood was precipitated with 380 μL methanol. The sample was then vortexed at 1000 rpm for 10 seconds and centrifuged at 3000 rpm for 10 minutes at 10 °C. 50 μL of the supernatant was removed and diluted to 1 mL with water. The sample was then vortexed and subjected to LC-MS/MS analysis. (Internal standard not shown on chromatogram.)

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