摘要/Abstract

摘要： 目的·建立人血浆中7α- 羟基 -4- 胆固醇烯 -3- 酮（7α-hydroxy-4-cholesten-3-one，C4）以及胆酸（cholic acid，CA）、鹅去氧胆酸（chenodeoxycholic acid，CDCA）的定量方法。方法·采用乙腈提取人血浆中C4、CA、CDCA，利用超高效液相色谱 - 串联质谱（ultra-high performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS）技术，通过内标校正的标准曲线法定量C4、CA、CDCA含量。结果·经方法学验证，C4、CA、CDCA的检出限（limits of detection，LOD)分别为0.16、0.02、0.04 nmol/L，均具有良好的线性关系（均R2>0.998）；重复性的相对标准偏差（relative standard deviation，RSD）均低于3.0%；日内及日间精密度的RSD均小于6%；4 ℃保存7 d结果稳定，RSD均小于10%；C4、CA和CDCA的平均加标回收率分别为97.7%、113.3%、105.0%。结论·该方法具有较高的检测灵敏度、良好的精密度和稳定性，可用于血浆生物样本的定量检测。
关键词: 7&, alpha, - 羟基 -4- 胆固醇烯 -3- 酮, 胆酸, 鹅去氧胆酸, 超高效液相色谱 - 串联质谱, 定量
Abstract:
Objective · To develop a quantitative method of 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA) and chenodeoxycholic acid (CDCA) in human plasma. Methods · After extraction of C4, CA and CDCA with acetonitrile plasma, they were quantified with standard curve correctedthe internal standards based on Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Results · The limits of detection (LOD) of C4, CA and CDCA were 0.16, 0.02 and 0.04 nmol/L respectively; All three metabolites had good linear relationships (correlation coefficients R2 were over than 0.998). The relative standard deviations (RSDs) of repeatabilities were below 3.0%. The RSDs of inter-day and intra-day precision were less than 6%, and the RSDs of stabilities at 4 °C were below 10% within 7 days. The average added recoveries of C4, CA and CDCA were 97.7%, 113.3% and 105.0%, respectively. Conclusion · This method is of high detective sensitivity, good precision and stability, which meets the quantitative requirements of plasma biological samples.
Key words: 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA), chenodeoxycholic acid (CDCA), UPLC-MS/MS, quantification


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