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Tetracyclines in Meat by LC/MS/MS

Sample Preparation Details

Part No:8B-S100-UBJ
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 60 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:Measure 5 g of sample into an 85 mL cenrifuge tube. Add 25 mL of acidified methanol/1M HCl mixture (0.833 mL of 1M HCl in a total volume of 200 mL methanol). Mix for 1 min using an Ultra-Turrax mixer at a velocity setting of 2. Agitate further for 5 min using an orbital mixer at a velocity setting of 100. Centrifuge at 4400 rpm, 10 C for 15 min. Filter 5 mL of the top liquid layer through a 45 um Phenex filter. Dilute 0.25 mL of the filtrate with 0.1 % formic acid in 4.75 mL of water. Filter through a 45 um Phenex filter.
Dry:Evaporate to dryness using an eppendorf concentrator set at 45C or blow dry with nitrogen.
Final Prep and Analysis: Inject: 50 µL on HPLC / Tandem Mass Spec (MS-MS) @ (600 °C)

Analytes:

Name Spiked
Conc.
Log P pKa % Rec %RSC (n=0)
1 Oxytetracycline
2 Tetracycline
3 Chlortetracycline
4 Methacycline
5 Doxycycline

Application Detail (App ID: 20032)

Part No:00B-4435-E0
Dimensions:50 x 4.6 mm ID
Elution Type:Gradient
Elution A:0.1% Formic Acid in Water
Elution B:Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 98 2
2 2 98 2
3 2.1 85 15
4 6.5 50 50
5 7.1 0 100
6 8 0 100
7 8.1 98 2
8 10 98 2

Flow Rate:0.8 mL/min
Col. Temp: 40 °C
Detection: Tandem Mass Spec (MS-MS) @ (600°C)
Detector Info: 3200 QTRAP LC/MS/MS
Note:SecurityGuard™ Guard Cartridge System extends column lifetime.
- SecurityGuard Cartridges, Gemini C18 4 x 3.0mm ID, 10/Pk Part No.: AJ0-7597
- Holder Part No.: KJ0-4282

Analyte Details

Analyte Detail not available

1
Oxytetracycline
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2
Tetracycline
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3
Chlortetracycline
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4
Methacycline
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5
Doxycycline
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