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Extraction of Phenylbutazone from Beef Using Strata-X-A on Kinetex 2.6u XB-C18 50x2.1 by LC/MS/MS

Sample Preparation Details

Condition 1 :3 mL Methanol at 1 mL/min
Condition 2 :3 mL Water at 1 mL/min
Load :Load 2.00 mL Sample
Wash 1 :2 mL Water at 1 mL/min
Wash 2 :2 mL Acetonitrile at 1 mL/min
Wash 3 :2 mL Ethylacetate at 1 mL/min
Dry :5 mins at 10" of Hg
Elution 1 :3 mL 1% formic acid/methanol at 1 mL/min
Evaporate: :Evaporate to dryness
Reconstitute :500 µL Initial mobile phase (A/B=50/50)
Notes :Extraction Procedure: 1. Condition a Strata-X-A, 100 mg/6 mL with 3 mL 100% MeOH followed by 3 mL DI water 2. Load sample from the final step of pretreatment section 3. Wash with 2 mL DI H2O 4. Wash with 2 mL 100% ACN 5. Wash with 2 mL 100% Ethylacetate 6. Dry the SPE bed for 4-5 min @ max vacuum 7. Elute with 2x1.5 mL 1% formic acid in MeOH 8. Evaporate the eluate to dryness under gentle nitrogen stream at 45-50 C 9. Re-suspend the residue with 500 uL 50:50 MeOH:0.1% formic acid in DI H2O

Application Detail (App ID: 21872)

Part No:00B-4496-AN
Dimensions:50 x 2.1 mm ID
Elution Type:Gradient
Elution A:0.1% Formic Acid in DI Water
Elution B:0.1% Formic Acid in Methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 50 50
2 0.5 50 50
3 2 5 95
4 3.5 5 95
5 3.51 50 50
6 5 50 50

Flow Rate:0.45 mL/min
Col. Temp: 45 °C
Detection: Mass Spectrometer (MS) @ amu (0°C)
Detector Info: AB Sciex API 4000
Note:Mass spec conditions:
API 4000, ESI TurboIon Spray, + Ionization;
CAD: 6.00
CUR: 20.00
GS1: 50.00
GS2: 50.00
IS: 5500.00
TEM: 600.00
ihe: ON
DP 50.00
EP 10.00


Analyte Details

Analyte Detail not available

1
Phenylbutazone
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