Clinica chimica acta; international journal of clinical chemistry · 2024
Application of magnetic bead-assisted liquid chromatography-tandem mass spectrometry method for analyzing serum bile acid profiles: Advancing toward automated mass spectrometry.
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Zhou Yu, Du Jing, Liu Yamei, Ma Jinfei, Yuan Xiaofen, Xia Jun
Аннотация
BACKGROUND: Magnetic bead-assisted extraction technology offers potential for automating mass spectrometry processes, yet research in this area remains limited. Here, we developed and validated a magnetic bead extraction method to simultaneously analyze 13 bile acid profiles in human serum.
METHODS: A magnetic bead-assisted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for serum bile acid profiles. The linearity, accuracy, precision, matrix effect, carryover, stability test and interference test of the LC-MS/MS method were assessed. Comparative analysis involving 20 pregnant women evaluated different methodologies for diagnosing and monitoring intrahepatic cholestasis of pregnancy (ICP).
RESULTS: The magnetic bead extraction method demonstrated analytical performance comparable to the traditional protein precipitation method. The method exhibited linear responses across the clinical reference interval for all 13 bile acids. Meanwhile, the recovery rates for bile acid analytes ranged from 85.14 to 114.43%, and the repeatability was within 1.68 to 10.83%, meeting acceptance criteria. Our magnetic bead-assisted extraction method showed no obvious matrix effects and carryover effects. The extracted specimens remained stable in the autosampler for 24 h, and the interference test suggested the bile acid profile results would not be significantly affected under different interfering substances. Furthermore, comparative studies with traditional protein precipitation method showed comparable results, and our method exhibited robust clinical consistency in monitoring the efficacy of ICP patients.
CONCLUSIONS: We developed a straightforward magnetic bead extraction method coupled with LC-MS/MS for bile acid profiles. It can be applied to the clinical routine diagnosis of ICP and facilitate the automated mass spectrometry processes.
METHODS: A magnetic bead-assisted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for serum bile acid profiles. The linearity, accuracy, precision, matrix effect, carryover, stability test and interference test of the LC-MS/MS method were assessed. Comparative analysis involving 20 pregnant women evaluated different methodologies for diagnosing and monitoring intrahepatic cholestasis of pregnancy (ICP).
RESULTS: The magnetic bead extraction method demonstrated analytical performance comparable to the traditional protein precipitation method. The method exhibited linear responses across the clinical reference interval for all 13 bile acids. Meanwhile, the recovery rates for bile acid analytes ranged from 85.14 to 114.43%, and the repeatability was within 1.68 to 10.83%, meeting acceptance criteria. Our magnetic bead-assisted extraction method showed no obvious matrix effects and carryover effects. The extracted specimens remained stable in the autosampler for 24 h, and the interference test suggested the bile acid profile results would not be significantly affected under different interfering substances. Furthermore, comparative studies with traditional protein precipitation method showed comparable results, and our method exhibited robust clinical consistency in monitoring the efficacy of ICP patients.
CONCLUSIONS: We developed a straightforward magnetic bead extraction method coupled with LC-MS/MS for bile acid profiles. It can be applied to the clinical routine diagnosis of ICP and facilitate the automated mass spectrometry processes.