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