Determination of oxysterols in human serum by liquid chromatography-silver coordination ion spray/tandem mass spectrometry(LC-AG+CIS/MS/MS)
- Determination of oxysterols in human serum by liquid chromatography-silver coordination ion spray/tandem mass spectrometry(LC-AG+CIS/MS/MS)
- 차은주; 김소희; 김희원; 김호준; 손정현; 권오승; 이재익
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- Oxysterols are mono-oxygenated forms of cholesterol and play an important role for preserving lipid homeostasis in the body. Recently, several study for cholesterols in biological samples were reported using liquid chromatography-tandem mass spectrometry equipped electrospray ionization or atmospheric pressure chemical ionization instead of the gas chromatography-mass spectrometry. However the lacks of ionization sites of cholesterols have restricted the sensitivity and reproducibility of analytical method. Therefore, we have developed a highly sensitive and specific method using Ag+ coordination ion spray ionization (CIS) to analyze the oxysterols such as 24S-hydroxycholesterol (24S-OHC), 25-hydroxycholesterol (25-OHC) or 27-hydroxycholesterol (27-OHC) in human serum. Calibration curves were obtained over a linear range of 5 to 250 ng/mL for 24S-OHC and 27-OHC, and 0.5 to 25 ng/mL for 25-OHC. Intra- and inter-day precisions were between 1.0% and 13.3% and the accuracies ranged from 94.3% to 109.0%. Limit of detection and limit of quantification were 0.3 ng/mL, 5 ng/mL for 24S-OHC and 27-OHC, and 0.03 ng/mL and 0.5 ng/mL for 25-OHC, respectively. In conclusion, we have validated the developed method for linearity, accuracy and precision and performed microanalysis for metabolites of cholesterol in human serum by a novel Ag+CIS method provides reproducibility and reliability for qualitative and quantitative analysis.
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