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Peaks | tR (min) | Precursor Ion | Product Ion | |
---|---|---|---|---|
1. | Nornicotine | 1.25 | 149.10 | 80.05 |
2. | Norcotinine | 1.81 | 163.09 | 80.05 |
3. | trans-3'-Hydroxycotinine | 1.88 | 193.12 | 80.05 |
4. | Nicotine | 1.95 | 163.15 | 132.10 |
5. | Anabasine | 2.20 | 163.13 | 91.63 |
6. | Cotinine | 2.88 | 177.12 | 98.07 |
Column | Raptor Biphenyl (cat.# 9309512) | ||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Dimensions: | 100 mm x 2.1 mm ID | ||||||||||||||||||||||||||||
Particle Size: | 5 µm | ||||||||||||||||||||||||||||
Pore Size: | 90 Å | ||||||||||||||||||||||||||||
Guard Column: | Raptor Biphenyl EXP cartridges 5 mm, 2.1 mm ID, 5 µm (cat.# 930950252) | ||||||||||||||||||||||||||||
Temp.: | 30 °C | ||||||||||||||||||||||||||||
Standard/Sample | |||||||||||||||||||||||||||||
Diluent: | Water | ||||||||||||||||||||||||||||
Conc.: | 25 ng/mL (fortified human urine) | ||||||||||||||||||||||||||||
Inj. Vol.: | 5 µL | ||||||||||||||||||||||||||||
Mobile Phase | |||||||||||||||||||||||||||||
A: | 0.1% Formic acid, 5 mM ammonium formate in water | ||||||||||||||||||||||||||||
B: | 0.1% Formic acid in methanol | ||||||||||||||||||||||||||||
|
Detector | MS/MS |
---|---|
Ion Mode: | ESI+ |
Mode: | MRM |
Instrument | UHPLC |
Sample Preparation | The urine from a non-tobacco user was fortified with analytes to a final concentration of 25 ng/mL and prepared by a liquid-liquid extraction method. A 250 μL urine sample was mixed with 50 μL of 5N NaOH and extracted with 1.5 mL of 50/50 methylene chloride/diethyl ether. A portion of organic phase (1.0 mL) was dried and reconstituted with 200 μL of water for analysis. |
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