Determination of β-Agonist Drugs in Animal Tissue

Method: SPE
Matrix: Animal tissue
Application No.: 101732
Columns: ProElut PXC 60 mg / 3 mL 50/pk
Cat No.: 68203
Sample Pretreatment: 1. Sample preparation
Weigh 5 g sample, add 15 mL ethyl acetate and 3 mL 10% sodium carbonate, homogenize at 10,000 rpm for 2 min, centrifuge at 6,000 rpm for 2 min, transfer the supernatant to the centrifuge tube, extract residues using 15 mL ethyl acetate, combine ethyl acetate layers. Add 5 mL 0.1 M hydrochloric acid to the ethyl acetate extract, vortex 1 min, centrifuge 1 min at 6,000 rpm, collect the lower aqueous phase. Repeat the extraction and combine lower aqueous phase, adjust to pH 5.2 with 2.5 mol/L sodium hydroxide.
2. Sample purification
ProElut™ PXC 60 mg / 3 mL (Cat#68203)
Condition: 3 mL MeOH / 3 mL H2O / 3 mL 30 mM HCl
Load: Add sample
Wash 1: 3 mL deionized water
Wash 2: 3 mL MeOH
Elute: 5 mL MeOH (4% NH4OH), evaporate to near dry at 50ºC by N2
3. Derivatization
Add 100 µL toluene and 100 µL bis-trimethylsilyl trifluoroacetamide (BSTFA), vortex 20 sec, seal, heat for 1 h at 80 ºC, add 300 µL of toluene after cooling as the sample solution.
Conditions: GC-MS method
GC conditions
Column: DM-5MS 30 m x 0.25 mm x 0.25 µm (Cat#8221)
Inlet Temperature: 220 ºC
Injection Mode: Splitless
Injection Volume: 1 µL
Temperature Program: Heating to 70 ºC in 0.6 min, then heating to 200 ºC with 25 ºC/min in 6 min, finally heating to 280 ºC with 25 ºC/min in 5 min
Carrier Gas: He > 99.999%, flow rate: 0.9 mL/min

MS conditions
Interface Temperature: 280 ºC
Solvent Delay: 8 min
EI Temperature: 230 ºC
Quadrupole Temperature: 160 ºC
Ion Monitoring: Qualitative ion 86, 243, 262, 277, quantitative ion 86
Author: Dikma Technologies Inc.
Keyword: β-Agonist Drugs, Animal Tissue, SPE, PXC