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采用液相色谱-串联质谱法测定血清中的醛固酮。

Determination of aldosterone in serum by liquid chromatography-tandem mass spectrometry.

作者信息

Turpeinen Ursula, Hämäläinen Esa, Stenman Ulf-Håkan

机构信息

HUSLAB, Laboratory of Women's Clinic, P.O. Box 140, 00029 HUS, Helsinki, Finland.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Feb 1;862(1-2):113-8. doi: 10.1016/j.jchromb.2007.11.005. Epub 2007 Nov 17.

Abstract

Measurement of serum aldosterone is clinically important in the diagnosis of hypertension. While isotope dilution gas chromatography-mass spectrometry (ID-GC-MS) provides reliable results, it requires derivatization and is lengthy and time-consuming. Detection by liquid chromatography-mass spectrometry (LC-MS) is a potentially superior method. The analysis utilizes 0.5mL of serum. The samples were extracted with dichloromethane-ether. The extract was evaporated to dryness and aldosterone was analyzed by LC-MS/MS operating in the negative mode ESI after separation on a reversed-phase column. Aldosterone was also measured by RIA. The calibration curves for analysis of serum aldosterone exhibited consistent linearity and reproducibility in the range of 60-3000pmol/L. Interassay CVs were 4.3-7.5% at aldosterone concentrations of 97-993pmol/L. The lower limit of quantitation (LOQ) was 30pmol/L (signal to noise ratio=10). The mean recovery of the analyte added to serum ranged from 95 to 102%. The regression equation by LC-MS/MS (x) and RIA (y) method was: y=1.33x+185 (r=0.95; n=124). Sensitivity and specificity of the LC-MS/MS method for serum aldosterone offer advantages over GC-MS by eliminating derivatization. The novel method is rapid, reliable and simple to perform with a routine LC-MS/MS spectrometer. The sensitivity is adequate for patient samples. Aldosterone concentrations reported by nonextraction RIA were consistently higher than those produced by LC-MS/MS.

摘要

血清醛固酮的测定在高血压诊断中具有重要临床意义。虽然同位素稀释气相色谱 - 质谱联用技术(ID - GC - MS)能提供可靠结果,但它需要进行衍生化处理,且耗时较长。液相色谱 - 质谱联用技术(LC - MS)检测是一种潜在的更优方法。该分析使用0.5mL血清。样品用二氯甲烷 - 乙醚萃取。提取物蒸发至干,在反相柱上分离后,通过在负离子模式电喷雾电离(ESI)下运行的LC - MS/MS分析醛固酮。醛固酮也通过放射免疫分析法(RIA)进行测定。血清醛固酮分析的校准曲线在60 - 3000pmol/L范围内呈现出一致的线性和可重复性。在醛固酮浓度为97 - 993pmol/L时,批间变异系数为4.3 - 7.5%。定量下限(LOQ)为30pmol/L(信噪比 = 10)。添加到血清中的分析物的平均回收率在95%至102%之间。LC - MS/MS(x)和RIA(y)方法的回归方程为:y = 1.33x + 185(r = 0.95;n = 124)。LC - MS/MS法检测血清醛固酮的灵敏度和特异性通过消除衍生化处理优于GC - MS法。这种新方法快速、可靠,使用常规LC - MS/MS光谱仪操作简单。其灵敏度对于患者样本来说足够。非萃取RIA法报告的醛固酮浓度始终高于LC - MS/MS法测定的浓度。

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