Potential of sterol analysis by liquid chromatography-tandem mass spectrometry for the prenatal diagnosis of Smith-Lemli-Opitz syndrome.
Potential of sterol analysis by liquid chromatography-tandem mass spectrometry for the prenatal diagnosis of Smith-Lemli-Opitz syndrome.
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DOI：
10.1373/clinchem.2007.100644
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发表时间：
2008-08
影响因子：
9.3
通讯作者：
Shackleton, Cedric
中科院分区：
文献类型：
--
作者：
Griffiths, William J.;Wang, Yuqin;Karu, Kersti;Samuel, Emmanuel;McDonnell, Shane;Hornshaw, Martin;Shackleton, Cedric
Smith-Lemli-Opitz综合征（SLOS）是一种严重的胆固醇合成障碍，通常通过羊水中固醇的气相色谱-质谱分析在产前诊断。鉴于目前串联质谱（MS/MS）方法的发展趋势，我们提出了用于准确诊断羊水中3β-羟基甾醇被胆固醇氧化酶氧化成相应的3-酮，然后在“一锅”反应中与吉拉德P （GP）肼衍生化的原型LC-MS/MS方法。由于带电季氮的存在，所得gp -腙在电喷雾(ES)-MS/MS中具有更好的响应，并通过反相LC-ES-MS/MS进行分析。毛细管LC-MS/MS和常规LC-MS/MS格式均适用，该方法也适用于纸吸收血斑。在一个双盲分析18羊水样本包括6 SLOS和12控制,7 + 8-dehydrocholesterol (7 + 8-DHC)胆固醇比被发现躺低于0.02(范围0.00 - 0.02:意味着±SD, 0.01±0.007)在所有控制样品(内部分析变异5.91%),0.20及以上(范围0.20 - 1.13:意味着±SD, 0.79±0.35)在SLOS(内部分析变异4.56%),对应两组之间的差异比率至少10倍。定量限相当于柱上注射2nl羊水。在这里，我们描述了一个“概念验证”的研究产前诊断的sls。然而，进一步的发展将是必要的自动化样品处理和减少色谱时间，以便使该方法用于产前和产后诊断。
Smith-Lemli-Opitz syndrome (SLOS) is a severe disorder of cholesterol synthesis classically diagnosed prenatally by GC-MS analysis of sterols in amniotic fluid. In recognition of the current move towards tandem mass spectrometry (MS/MS) methodologies we present prototype LC-MS/MS methods for the accurate diagnosis of the disorder 3β-Hydroxysterols in amniotic fluid are oxidised with cholesterol oxidase to their corresponding 3-ketones, which are then derivatised with Girard P (GP) hydrazine in a “one-pot” reaction. The resulting GP-hydrazones give an improved response in electrospray (ES)-MS/MS due to the presence of a charged quaternary nitrogen and are analysed by reversed-phase LC-ES-MS/MS. Both capillary LC-MS/MS and conventional LC-MS/MS formats are suitable, and the method is also applicable to paper absorbed blood spots. In a double blind analysis of 18 amniotic fluid samples comprising 6 SLOS and 12 controls, the 7+8-dehydrocholesterol (7+8-DHC) to cholesterol ratio was found to lie below 0.02 (range, 0.00 - 0.02: mean ± SD, 0.01 ± 0.007) in all control samples (intra assay variation 5.91%), and above 0.20 (range, 0.20 - 1.13: mean ± SD, 0.79 ± 0.35) in SLOS (intra assay variation 4.56%), corresponding to a difference in ratios between the two groups of a factor of at least 10. The limit of quantification was equivalent to 2 nL of amniotic fluid injected on-column. Here we describe a “proof-of-concept” study for the prenatal diagnosis of SLOS. However, further developments will be necessary to automate sample handling and reduce chromatographic time in order to allow the methodology to be used for pre- and postnatal diagnosis.
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DOI：
10.1002/ajmg.1320560309
发表时间：
1995-04-10
期刊：
AMERICAN JOURNAL OF MEDICAL GENETICS
影响因子：
--
作者：
ABUELO, DN;TINT, GS;SALEN, G
通讯作者：
SALEN, G
影响因子：
5.2
作者：
Starck, L;Lövgren, A
通讯作者：
Lövgren, A
DOI：
10.1016/j.jasms.2005.10.012
发表时间：
2006-03-01
影响因子：
3.2
作者：
Griffiths, WJ;Wang, YQ;Sjövall, J
通讯作者：
Sjövall, J
影响因子：
3
作者：
Chevy, F;Humbert, L;Wolf, C
通讯作者：
Wolf, C
影响因子：
158.5
作者：
TINT, GS;IRONS, M;SALEN, G
通讯作者：
SALEN, G