Determination of Sulfonamides in Animal Tissue

Method: SPE
Matrix: Animal tissue
Application No.: 101726
Columns: ProElut PLS 60 mg / 3 mL 50/pk
Cat No.: 68003
Sample Pretreatment: 1. Sample preparation
Weigh 5 g sample, add 5 g anhydrous sodium sulfate and 25 mL ethyl acetate, homogenize at 10,000 rpm for 2 min,centrifuge at 4,000 rpm for 2 min, collect ethyl acetate layer. Repeat 25 mL ethyl acetate extraction, combined ethyl acetate extracts, and vacuum distillation at 30 ºC to near dry. Add 1 mL methanol, 2 mL 1% acetic acid and 3 mL n-hexane to the distillation flask, vortex 1 min, then transfer to 15 mL centrifuge tube. Repeat the dissolution process, add mixture to the centrifuge tube, vortex 1 min, centrifuge 1 min at 4,000 rpm, discard the hexane. Add 6 mL n-hexane and repeat the operation. Finally, add 6 mL deionized water to the lower layer.
2. Sample purification
ProElut™ PLS 60 mg / 3mL (Cat#68003 )
Condition: 3 mL MeOH / 3 mL H2O
Load: Add sample
Wash 1: 3 mL H2O
Wash 2: 3 mL MeOH:H2O = 5:95
Elute: 5 mL MeOH
Reconstitute: Evaporate at 30 ºC by N2, reconstitute to 1 mL with mobile phase
Conditions: HPLC method
Column: Inspire™ 5 µm C18, 250 x 4.6 mm (Cat#81006)
Mobile Phase: A: MeCN, B: 2 % CH3COOH in H2O
Flow Rate: 1.0 mL/min
Temperature: 35 ºC
Detection: UV 270 nm
Injection Volume: 20 µL
Author: Dikma Technologies Inc.
Keyword: Sulfonamides, Animal Tissue, SPE, PLS