LC/MS with Online Solid Phase Extraction (SPE-LC/MS) of Thyroid Hormones in Plasma on Ascentis® Express Phenyl-Hexyl using RP-Amide Traps
材料
used together
相关产品
L-甲状腺素（T4） 溶液
100 μg/mL in methanol with 0.1N NH3, ampule of 1 mL, certified reference material, Cerilliant®3,3′,5-三碘-L-甲状腺原氨酸（T3） 溶液
100 μg/mL in methanol with 0.1N NH3, ampule of 1 mL, certified reference material, Cerilliant®3,3′,5′-三碘代-甲腺原氨酸（rT3）标准液 溶液
100 μg/mL in methanol with 0.1N NH3, ampule of 1 mL, certified reference material, Cerilliant®CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
column
Ascentis Express Phenyl-Hexyl, 5 cm x 2.1 mm I.D., 2.7 μm particles (53334-U)
mobile phase
[A] water; [B] methanol, both with 0.1% acetic acid
gradient
5% B for 3 min; to 70% B in 2 min, hold 3 min
pressure
1800 psi
column temp.
ambient
detector
MS, ESI(+), MRM mode (777.7/731.8 for T4; 651.8/605.5 for T3 or rT3)
injection
5 μL
sample
protein precipitated rabbit plasma spiked with desired amount of T4, T3 and rT3
说明
分析说明
An online SPE-LC/MS method is shown here for the determination of thyroid hormones in blood plasma. The method exploits RP-Amide or C8 as the trapping column and a phenyl-hexyl phase as the analytical column, respectively. Under the optimized conditions, both RP-Amide and C8 effectively trapped the thyroid hormones extracted from spiked rabbit plasma sample which had been protein precipitated. However, the RP-Amide traps were shown to be superior to the commonly used C8 traps because they provided higher analyte signals and recoveries. Additionally, RP-Amide traps are more flexible with washing solvent as they are compatible with 100% aqueous mobile phases and can be used with up to 20% methanol with minimal sample loss. Separation was carried out using an Ascentis Express 2.7 μm Phenyl-Hexyl column. The highest grade solvents provided clean, robust operation. Cerilliant CRMs provided reliable quantification.
法律信息
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany