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여름정기학술대회

2022여름초록

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Development and validation of a highly sensitive method for quantification of oxysterols as picolinyl ester derivatives using LC-ESI/MS/MS in serum

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대구경북첨단의료산업진흥재단
발표구분
포스터발표
포스터발표
Medical/Pharmaceutical Science
Brief Oral Presentation 발표신청
Keyword
Oxysterol
Liquid Chromatography
Mass Spectrometry
Picolinyl ester derivatization
Serum

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대구경북첨단의료산업진흥재단
국가
대한민국

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대구경북첨단의료산업진흥재단
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대한민국
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국가
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대구경북첨단의료산업진흥재단
To analysis of oxysterols as oxidized cholesterol is important and is highly challenging due to their low concentration in biological fluids. Thus, highly sensitive and selective analytical method is essential for the simultaneous quantitative determination of the hydroxycholesterols (OHCs) concentration. In this study, we developed analytical method to overcome the bottleneck via picolinyl ester (PE) derivatization, and validated for six OHCs including 24S-OHC, 25-OHC, 27- OHC, 24-diOHC, 25-diOHC, and 27-diOHCs in serum. Calibration curves were obtained over a linear range of 0.1 to 250 ng/mL for oxysterols except 25-OHC and 0.5 to 250 ng/mL for 25-OHC. Intra- and inter-day precisions were between 1.0% and 9.8% and the accuracies ranged from 87.6% to 114.2%. Limit of detection and lower limit of quantification were 0.05 ng/mL and 0.1 ng/mL for oxysterols except 25-OHC, and 0.1 ng/mL and 0.5 ng/mL for 25-OHC, respectively. Processed sample stabilities and recoveries were obtained in range of 86.3% to 116.8% and 32.5% to 107.6%, respectively. Pooled serums from obtained different batches were determined for oxysterol analysis. Profiles of oxysterols in serum were successfully applied and evaluated. OHCs spiked serums for potential biomarker discovery were preliminarily analyzed. In conclusion, we presented a highly sensitive method for quantification of oxysterols in human serum by LC-ESI/MS/MS with PE derivatization. This method is powerful tool and provides reproducibility and reliability for simultaneous analysis of oxysterols. * This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2020R1F1A1071199).

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