Synthetic Cannabinoid Metabolites on Ultra Biphenyl (500 ng/mL Extracted Urine Standard)
| Peaks | RT (min) | MRM1 | MRM2 | MRM3 | |
|---|---|---|---|---|---|
| 1. | JWH-073 4-hydroxybutyl | 2.05 | 344.1/155.1 | 344.1/127.2 | 344.1/144.0 |
| 2. | JWH-073 N-butanoic acid | 2.14 | 358.1/155.1 | 358.1/127.3 | 358.1/144.1 |
| 3. | JWH-018 N-pentanoic acid-d4 (IS) | 2.52 | 376.1/155.2 | 376.1/230.4 | 376.1/248.3 |
| 4. | JWH-018 N-pentanoic acid | 2.58 | 372.1/155.2 | 372.1/127.1 | 372.1/144.1 |
| 5. | JWH-018 5-hydroxypentyl | 2.58 | 358.1/155.2 | 358.1/127.1 | 358.1/230.3 |
| 6. | JWH-073 6-hydroxyindole | 3.53 | 344.1/155.2 | 344.1/127.1 | 344.1/145.1 |
| 7. | JWH-073 5-hydroxyindole | 3.69 | 344.1/155.2 | 344.1/127.1 | 344.1/160.0 |
| 8. | JWH-073 7-hydroxyindole | 3.95 | 344.2/155.1 | 344.2/127.1 | 344.2/216.3 |
| 9. | JWH-018 6-hydroxyindole | 4.00 | 358.1/155.1 | 358.1/127.2 | 358.1/145.2 |
| 10. | JWH-018 5-hydroxyindole | 4.13 | 358.1/155.1 | 358.1/127.2 | 358.1/160.2 |
| 11. | JWH-018 7-hydroxyindole | 4.34 | 358.1/155.1 | 358.1/127.1 | 358.1/230.3 |
| 12. | JWH-073 4-hydroxyindole-d7 (IS) | 5.11 | 351.3/127.0 | 351.3/223.0 | 351.3/155.0 |
| 13. | JWH-073 4-hydroxyindole | 5.15 | 344.1/155.2 | 344.1/127.2 | 344.1/160.0 |
| 14. | JWH-018 4-hydroxyindole | 5.44 | 358.1/155.2 | 358.1/230.2 | 358.1/127.1 |
LC_CF0532
| Column | Ultra Biphenyl (cat.# 9109552) | ||||||||||||||||||||||||||||||||
| Dimensions: | 50 mm x 2.1 mm ID | ||||||||||||||||||||||||||||||||
| Particle Size: | 5 µm | ||||||||||||||||||||||||||||||||
| Pore Size: | 100 Å | ||||||||||||||||||||||||||||||||
| Temp.: | 25 °C | ||||||||||||||||||||||||||||||||
| Sample | |||||||||||||||||||||||||||||||||
| Diluent: | 50:50 mobile phase | ||||||||||||||||||||||||||||||||
| Conc.: | 500 ng/mL extracted spiked sample | ||||||||||||||||||||||||||||||||
| Inj. Vol.: | 10 µL | ||||||||||||||||||||||||||||||||
| Mobile Phase | |||||||||||||||||||||||||||||||||
| A: | water + 0.05% acetic acid (pH approx. 3.4) | ||||||||||||||||||||||||||||||||
| B: | acetonitrile + 0.05% acetic acid | ||||||||||||||||||||||||||||||||
| |||||||||||||||||||||||||||||||||
| Detector | API 4000 | ||||||||||||||||||||||||||||||||
| Model #: | API 4000 | ||||||||||||||||||||||||||||||||
| Ion Source: | TurboIonSpray® | ||||||||||||||||||||||||||||||||
| Ion Mode: | ESI+ | ||||||||||||||||||||||||||||||||
| Ion Spray Voltage: | 3 kV | ||||||||||||||||||||||||||||||||
| Curtain Gas: | 40 psi (275.8 kPa) | ||||||||||||||||||||||||||||||||
| Gas 1: | 40 psi (275.8 kPa) | ||||||||||||||||||||||||||||||||
| Gas 2: | 40 psi (275.8 kPa) | ||||||||||||||||||||||||||||||||
| Interface Temp.: | 600 °C | ||||||||||||||||||||||||||||||||
| Mode: | MRM | ||||||||||||||||||||||||||||||||
| Dwell Time: | 30 ms | ||||||||||||||||||||||||||||||||
| Instrument | API LC-MS/MS | ||||||||||||||||||||||||||||||||
| Notes | Since multiple transitions are shared between analytes, only 5 transitions are shown to simplify viewing. The transitions shown are: 344.1/155.1 (blue trace), 358.1/155.1 (green trace), 372.1/155.2 (grey trace), 376.1/155.2 (purple trace - internal standard), 351.3/127.0 (red trace - internal standard) CAD Gas was set to 4 psi. For 1 ng/mL calibration level, see chromatogram LC_CF0530. For 50 ng/mL calibration level, see chromatogram LC_CF0531. Sample was prepared according to the following method: 1) Spike 1 mL blank urine sample with analytes and internal standards. 2) Hydrolyze sample: - Add 1 mL solution of beta-glucuronidase from keyhole limpet (Sigma-Aldrich cat.# G8132). Solution is prepared at a concentration of 5,000 Fishman units/mL in 100 mM ammonium acetate buffer (pH = 5.0). - Incubate at 60 °C for 3 hours. 3) Extract sample on 6 mL, 500 mg C18 high-load endcapped Resprep® SPE cartridge (cat.# 24052): - Add 1 mL 5 mM ammonium acetate + 0.1% acetic acid (pH = 4.2) to sample. - Condition cartridge with 3x 1 mL acetonitrile. - Condition cartridge with 3x 1 mL 5 mM ammonium acetate + 0.1% acetic acid. - Apply sample and allow to pass through under gravity. - Rinse with 3x 1 mL 5 mM ammonium acetate + 0.1% acetic acid. - Dry cartridge with vacuum for 10 minutes. - Elute with 3 mL acetonitrile followed by 3 mL butyl chloride. 4) Concentrate sample: - Evaporate sample to dryness under nitrogen at 40 °C. - Reconstitute in 0.5 mL water + 0.05% acetic acid:acetonitrile + 0.05% acetic acid (50:50). | ||||||||||||||||||||||||||||||||
| Acknowledgement | Special thanks to Cayman Chemical for reference standards |
