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Chiral Separation of Dextromethorphan/Levomethorphan and Dextrorphan/Levorphanol on Lux 3u AMP, 150x4.6mm

Sample Preparation Details

Part Number :8E-S029-TGB
Description :Strata™ X-C 33 µm Polymeric Strong Cation
Condition 1 :1 mL methanol
Sample Pretreatment :Urine sample pretreatment: Combine 200µL urine sample, 200µL 100mM ammonium acetate buffer (pH 4.0), 20µL internal standard solution (250ng/mL), and 20µL of beta-glucuronidase (>100,000 units/mL) and incubate for 1 hour at 55°C. Centrifuge for 10 minutes at 13,000rcf. Dilute 400µL supernatant 1:1 with 0.1% aqueous formic acid. Serum sample pretreatment: Combine 250µL serum, 250µL 0.1% formic acid, and 20µL internal standard solution (250 ng/mL).
Load :Full pretreated sample (from above)
Wash 1 :1.0% formic acid in water
Wash 2 :1.0% formic acid in methanol
Dry :3-4 minutes at high vacuum
Elution 1 :2 x 0.5 mL 5% NH4OH in 50:50 methanol/acetonitrile
Evaporate: :Evaporate to dryness
Reconstitute :100 µL 40:60 acetonitrile/water
Condition 1 mL methanol
Equilibrate 1 mL 0.1% formic acid in water
Load Full pretreated sample (from above)
Wash 1.0% formic acid in water
Wash 1.0% formic acid in methanol
Dry 3-4 minutes at high vacuum
Elute 2 x 0.5 mL 5% NH4OH in 50:50 methanol/acetonitrile
Dry Down at 40°C under a gentle stream of nitrogen
Reconstitute 100 µL 40:60 acetonitrile/water

Application Detail (App ID: 25783)

Part No:00F-4751-E0
Dimensions:150 x 4.6 mm ID
Elution Type:Gradient
Elution A:5mM ammonium bicarbonate, adjusted to pH 11 with ammonium hydroxide
Elution B:Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 55 45
2 4.5 55 45
3 4.6 20 80
4 8 20 80
5 8.1 55 45
6 10 55 45

Flow Rate:1 mL/min
Col. Temp: 40 °C
Detection: LC/MS/MS @ (0°C)
Note:Urine hydrolyzed with beta-glucuronidase and extracted with Strata-X-C

Analyte Details

Analyte Detail not available

1
Dextrorphan 1
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2
Dextrorphan 2
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3
Levorphanol 1
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4
Levorphanol 2
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5
Dextromethorphan 1
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6
Dextromethorphan 2
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7
Levomethorphan 1
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8
Levomethorphan 2
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