Chloramphenicol in Seafood and dairy products 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:Homogenize ~100 g of thawed shrimp using blender or tissue homogenizer. Weigh out 5 g of homogenized shrimp and transfer to a 15 mL polypropylene tube. Add 50 uL of d5-Chloramphenicol (IS) solution. Mix thoroughly using a vortex mixer to ensure adequate distribution of the IS throughout the homogenate. Add 2 mL water and mix well using a vortex mixer. Add 5 mL ethyl acetate and transfer the tubes to a mechanical shaker, shake rigorously for 30 minutes. Centrifuge at 7000 rpm for 15 min. Transfer the supernatant to a new 15 mL polypropylene test tube, reserving the pelleted shrimp homogenate tissue. Add 5 mL ethyl acetate to the pelleted shrimp homogenate tissue and repeat the extraction process. Combine the resulting ethyl acetate supernatant with the extraction previously obtained. Dry the combined supernatant extracts under nitrogen at 55 C. Reconstitute with 300 uL methanol and then dilute to 10 mL with water.
Dry:5 minutes @ 10 in Hg Vaccum
Final Prep and Analysis: Inject: 25 µL on HPLC / Tandem Mass Spec (MS-MS) @ (450 °C)


Name Spiked
Log P pKa % Rec %RSC (n=0)
1 Chloramphenicol
2 Chloramphenicol
3 Chloramphenicol-d5

Application Detail (App ID: 20026)

Part No:00B-4462-AN
Dimensions:50 x 2.1 mm ID
Elution Type:Gradient
Elution A:Water
Elution B:Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 -2 95 5
2 0 95 5
3 2 95 5
4 2.01 5 95
5 4.5 5 95

Flow Rate:0.4 mL/min
Col. Temp: 20 °C
Detection: Tandem Mass Spec (MS-MS) @ (450°C)
Detector Info: AB Sciex API 4000™
Note:SecurityGuard™ ULTRA Guard Cartridge System extends column lifetime.
- SecurityGuard ULTRA Cartridges, UHPLC C18 for 2.1mm ID Columns, 3/Pk Part No.: AJ0-8782
- Holder Part No.: AJ0-9000

Analyte Details

Analyte Detail not available