Determination of an Endogenous Biomarker - 4beta-Hydroxycholesterol in K_(2)EDTA Human Plasma by LC-MS/MS

摘要

1. A highly sensitive, selective, and rugged LC-MS/MS method was developed and validated using picolinic acid as the derivatization reagent. 2. The derivatization conditions were optimized and reaction was completed with 30 min incubation at RT. 3. Water was used as the surrogate for human plasma to prepare working standards since 4beta-HC is an endogenous compound. QCs were prepared in human plasma to mimic incurred samples. 4. 4beta-HC was completely separated from 8 commercially available isomers of 4beta-HC and 4alpha-HC which was synthesized in our lab. 5. Method validation data met the validation acceptance criteria defined in the method validation protocol.