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

LC_CF0732
PeakstR (min)Precursor ionProduct ion
1.γ-Hydroxybutyric acid (GHB)1.42105.287.0
2.1,4-Butanediol (1,4-BD)1.4291.055.0
3.γ-Butyrolactone (GBL)1.7587.045.0
ColumnForce 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
Sample
Diluent:Water
Conc.:500 ng/mL
Inj. Vol.:10 µL
Mobile Phase
A:0.5% Formic acid in water
B:0.5% Formic acid in methanol
Time (min)Flow (mL/min)%A%B
0.000.7955
0.500.7955
3.000.75050
3.010.7955
5.000.7955
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentHPLC
Notes100 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.)