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Peaks | tR (min) | Conc. (ng/mL) | Precursor Ion | Product Ion 1 | Product Ion 2 | |
---|---|---|---|---|---|---|
1. | EtG-d5 | 0.76 | 100 | 225.9 | 84.7 | - |
2. | EtG | 0.80 | 500 | 220.9 | 74.9 | 85 |
3. | EtS-d5 | 1.70 | 100 | 129.7 | 97.7 | - |
4. | EtS | 1.78 | 500 | 124.8 | 96.8 | 79.7 |
5. | Phenobarbital-d5 | 2.54 | 100 | 236.0 | 42.0 | - |
6. | Phenobarbital | 2.55 | 500 | 231.2 | 188.0 | 42.0 |
7. | Butalbital-d5 | 2.57 | 100 | 228 | 42.0 | - |
Peaks | tR (min) | Conc. (ng/mL) | Precursor Ion | Product Ion 1 | Product Ion 2 | |
---|---|---|---|---|---|---|
8. | Butalbital | 2.58 | 500 | 223 | 180.0 | 42.0 |
9. | Amobarbital-d5 | 2.74 | 100 | 230 | 42.0 | - |
10. | Pentobarbital-d5 | 2.74 | 100 | 230 | 42.0 | - |
11. | Amobarbital | 2.75 | 500 | 225 | 182.0 | 42.0 |
12. | Pentobarbital | 2.76 | 500 | 225 | 182.0 | 42.0 |
13. | Secobarbital-d5 | 2.93 | 100 | 242 | 42.0 | - |
14. | Secobarbital | 2.93 | 500 | 237 | 194.0 | 42.0 |
Column | Raptor EtG/EtS (cat.# 9325A12) | ||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Dimensions: | 100 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.# 25809) | ||||||||||||||||||||||||||||
Temp.: | 30 °C | ||||||||||||||||||||||||||||
Standard/Sample | Ethyl-ß-D-glucuronide (cat.# 34101) | ||||||||||||||||||||||||||||
Ethyl-ß-D-glucuronide-d5 (cat.# 34102) | |||||||||||||||||||||||||||||
Ethyl sulfate sodium salt (cat.# 34103) | |||||||||||||||||||||||||||||
Ethyl sulfate-d5 sodium salt (cat.# 34104) | |||||||||||||||||||||||||||||
Phenobarbital (cat.# 34037) | |||||||||||||||||||||||||||||
Pentobarbital (cat.# 34036) | |||||||||||||||||||||||||||||
Butalbital (cat.# 34032) | |||||||||||||||||||||||||||||
Secobarbital (cat.# 34038) | |||||||||||||||||||||||||||||
Diluent: | Water, 0.1% formic acid | ||||||||||||||||||||||||||||
Inj. Vol.: | 10 µL | ||||||||||||||||||||||||||||
Mobile Phase | |||||||||||||||||||||||||||||
A: | Water, 0.1% formic acid | ||||||||||||||||||||||||||||
B: | Acetonitrile, 0.1% formic acid | ||||||||||||||||||||||||||||
|
Detector | MS/MS |
---|---|
Ion Source: | Electrospray |
Ion Mode: | ESI- |
Sample Preparation | Human urine (alcohol free) was fortified with EtG, EtS, phenobarbital, butalbital, amobarbital, pentobarbital, and secobarbital in order to prepare the calibration standards and QC samples. 50 µL of urine was diluted with 950 µL of the working internal standard (100 ng/mL EtG-d5, EtS-d5, and barbiturates-d5 in 0.1% formic acid in water). Samples were vortexed at 3500 rpm for ten seconds to mix, centrifuged at 3000 rpm for five minutes at 10 °C, and injected for LC-MS/MS analysis. |
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