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- 저자 : 나승훈(Seonghoon Na) (검색결과 2 건)
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초고성능액체크로마토그래피-탠덤매스에 의한 혈장과 담즙 중 콜릭산의 정량 분석에 관한 연구
나승훈(Seonghoon Na),윤혜란(Hye-Ran Yoon) 대한약학회 2019 약학회지 Vol.63 No.6
Bile acids are increasingly appreciated as bioactive molecules and important end products of cholesterol metabolism. While they have been identified as key factors in lipid emulsification and absorption due to their detergent properties. bile acids have also been shown to act as signaling molecules and intermediates between the host and the gut microbiota. To investigate bile acid functions in humans, an advanced platform for high throughput analysis is essential. Herein, we developed the analytical method of cholic acids following simple one step protein precipitation from biological sample by UPLC-MS/MS. MRM ions was m/z=407.2 for cholic acid. The R2 of calibration curves provided 0.9995 in the calibration range of 0.005~5 μg/mL. The quantification system was validated with excellent sensitivity that allows quantitative targeted analyses of bile acids. The LOD (0.001 μg/mL), LOQ (0.005 μg/mL), recoveries (96.8~101.3%± 2.7~4.0%) on intra-day assay, and (98.4~111.0%±2.3~3.4%) on inter-day assay achieved resonable validated data for bile acids analyses. The developed method was applied for drawing plasma and bile acid profile in both normal and disease status. Our study is characterized by rapid and simple sample preparation as well as successful application to plasma and bile. These results could be usable for routine diagnostic monitoring of bile acids on human biofluids.
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초고성능액체크로마토그래피-텐덤질량분석법에 의한 혈청 중스핑고신과 스핑고신-1-포스페이트의 동시 분석
나승훈(Seonghoon Na),윤혜란(Hye-Ran Yoon) 대한약학회 2021 약학회지 Vol.65 No.1
Sphingosine (SPH) and sphingosine-1-phosphate (S1P) are emerging as key players in asthma metabolism and in numerous cellular inflammation processes. To identify potential biomarkers of asthma and inflammatory therapeutics, it is essential to determine their levels. Herein, we developed a rapid and sensitive UHPLC-MS/MS method to simultaneously quantify SPH and S1P in human serum using C17-SPH and C17-S1P as internal standards. After methanol precipitation of serum proteins, the supernatants were analyzed by MS/MS performed in the positive ion mode by multiple reaction monitoring. UHPLC analysis (C18 column) was performed using two mobile phase systems (water containing 0.1% formic acid, and 85% acetonitrile containing 0.1% formic acid) within 5 min of the short run. The calibration curves were linear in the range of 0.002-1.5 µg/mL for S1P and SPH with an R 2 greater than 0.9999. The LOD and LOQ were 0.0002 and 0.0004 µg/mL for S1P, and 0.0005 and 0.001 µg/mL for SPH, respectively. The accuracy and precision of the method were in the range of 89.8-100.7% (RSD, 1.5-2.8%) for both SPH and S1P species. We were able to quantify both molecules in serum from healthy and asthmatic patients. These results suggest that SPH and S1P are promising potential biomarkers, and also contribute to the basic data for the construction of an omics-based platform for preventive index prior to asthma diagnosis.
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