LC/MS/MS Analysis of Drugs in Plasma on Ascentis® Express C18 after Extraction using SPME LC Tips
材料
SPME 光纤
分析色谱柱
标准
普萘洛尔 盐酸盐 溶液
1.0 mg/mL in methanol (as free base), ampule of 1 mL, certified reference material, Cerilliant®CONDITIONS
sample preparation
Solid Phase Microextraction
sample/matrix
rat plasma stabilized with K2EDTA and spiked at 100 ng/mL with each analyte
SPME fiber
SPME LC Tips, C18 chemistry (57234-U)
extraction
immersion with agitation, 30 minutes
desorption process
acetonitrile, 10 minutes
column
Ascentis Express C18, 5 cm x 2.1 mm I.D., 2.7 μm particles (53822-U)
mobile phase
[A] 5 mM ammonium formate in water; [B] 5 mM ammonium formate in acetonitrile:water (90:10 )
gradient
5% to 70% B in 3 min, to 90%B in 0.1 min, held at 90%B for 0.9 min
flow rate
0.5 mL/min
pressure
4641 psi (320 bar)
column temp.
40 °C
detector
MS/MS ESI+, MRM m/z
injection
2 μL
说明
分析说明
Determination of free circulating drug is important in establishing the drug′s pharmacokinetic activity. In most cases, drug-protein complexes are formed that affect the active level of circulating drugs. In this study, biocompatible SPME was used as a rapid means of determining drug-protein binding affinities from plasma. HPLC separation was on an Ascentis Express C18 column packed with Fused-Core particles.
法律信息
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany