Determination of Tetracyclines in Animal Tissue

Method: SPE
Matrix: Animal tissue
Application No.: 101735
Columns: ProElut PLS 60 mg / 3 mL 50/pk
Cat No.: 68003
Sample Pretreatment: 1. Sample preparation
To 5.0 g of homogenized sample, add 20 mL of McIlvaine buffer*, vortex 2 min, centrifuge at 4000 rpm for 5 min, collect supernatant. Wash lower residue with 20 mL, and then 10 mL Mcllvaine buffer. Repeat wash and combine extracts, and set the volume to 50 mL. Filter extract with fast filter paper, collect filtrate and take 10 mL as the sample solution.
*McIlvaine buffer: Disodium hydrogen phosphate (Na2HPO4•12H2O) 27.6 g, citric acid (C6H8O7•H2O) 12.9 g,EDTA disodium salt 37.2 g, dissolved in water and diluted to 1,000 mL.
2. Sample purification
ProElut™ PLS 60 mg / 3 mL (Cat#68003)
Condition: 3 mL MeOH / 3 mL H2O
Load*: 10 mL sample
Wash 1: 3 mL H2O
Wash 2: 3 mL 5 % MeOH in H2O
Elute: 3 mL MeOH
Reconstitute: Evaporate to near dry, reconstitute to 1 mL with mobile phase
*20 mL reservoir (Cat#4811) and adaptor (Cat#4803) is avilable for large volume sample
Conditions: Column: Spursil™ 5 µm C18, 150 x 4.6 mm (Cat#82001)
Mobile Phase: A: 0.01 M Oxalic acid in H2O, B: MeOH:MeCN = 1:1
Flow Rate: 1.0 mL/min
Temperature: 30 ºC
Detection: UV 365 nm
Injection Volume: 20 µL
Author: Dikma Technologies Inc.
Keyword: Tetracyclines, Animal Tissue, SPE, PLS