Determination of Sulfonamides in Milk and Milk Powder

Method:	SPE
Matrix:	Milk
Application No.:	101728
Columns:	ProElut PXC 200 mg / 6 mL 30/pk
Cat No.:	68212
Sample Pretreatment:	1. Sample preparation To 15 mL milk (or 3 g milk powder in 15 mL H₂O), add 15 mL acetonitrile, vortex for 2 min, centrifuge at 6,000 rpm for 5 min, transfer 20 mL supernatant to another centrifuge tube, and add 15 mL n-hexane, vortex for 2 min, centrifuge at 6,000 rpm for 2 min, then discard the n-hexane. Repeat 15 mL n-hexane extraction. Add 15 mL ethyl acetate to the lower layer, vortex for 2 min, centrifuge at 6,000 rpm for 2 min, and collect supernatant. Repeat 15 mL ethyl acetate extraction, and combine supernatants. Vacuum evaporate the ethyl acetate layer to near dry at 30 ºC, reconstitute with 10 mL 2% phosphoric acid. 2. Sample purification ProElut™ PXC 200 mg / 6 mL (Cat#68212 ) Condition: 6 mL MeOH / 6 mL H₂O Load: Add sample Wash 1: 6 mL H₂O Wash 2: 6 mL MeOH Elute: 6 mL MeOH (5 % NH₄OH) Reconstitute: Vacuum evaporation at 30 ºC, 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 % CH₃ COOH 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, Milk, Milk Powder, SPE, PXC