Methods in endogenous steroid profiling - A comparison of gas chromatography mass spectrometry (GC-MS) with supercritical fluid chromatography tandem mass spectrometry (SFC-MS/MS)
Methods in endogenous steroid profiling - A comparison of gas chromatography mass spectrometry (GC-MS) with supercritical fluid chromatography tandem mass spectrometry (SFC-MS/MS)
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DOI：
10.1016/j.chroma.2018.04.035
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发表时间：
2018-06-15
影响因子：
4.1
通讯作者：
Parr, Maria Kristina
中科院分区：
文献类型：
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作者：
Teubel, Juliane;Wuest, Bernhard;Parr, Maria Kristina
在内分泌学的各个领域中，人体合成的类固醇激素的测定起着重要的作用。在过去的几年里，中枢神经类固醇的研究得到了加强，因为它们被讨论为各种认知障碍的生物标志物。它们在脑脊液（CSF）中的浓度被认为是独立于外周液体调节的。因此，具有挑战性的基质CSF成为一个非常有趣的分析样本。预计浓度很低，可用的脑脊液量有限。因此，需要一种综合的方法，对一个分析同物中尽可能多的一组分析物进行非常灵敏的定量分析。然而，51种类固醇和类固醇硫酸盐（包括许多异构体）的高结构相似性给色谱选择性的实现带来了挑战。几十年来，各种体液中内源性类固醇的分析主要是通过气相色谱（GC）耦合（串联）质谱（MS）进行的。由于感兴趣的类固醇的结构，进行衍生化以满足GC-MS（/MS）的分析要求。大多数实验室在多分析物分析中使用两步衍生法，这种方法早在20世纪80年代就已发表。然而，对于某些类固醇，这个复杂的过程产生多个同分异构体衍生物。因此，一些实验室使用（超）高效液相色谱((U)HPLC)-质谱/质谱作为替代，但是，即使是UHPLC也无法分离某些同分异构体对。超临界流体色谱（SFC）作为一种与气相色谱（GC）和(U)高效液相色谱（HPLC）相结合的正交分离技术，可能有助于克服这些问题。本项目对两种最有前途的内源性类固醇分析方法进行了研究和比较：“金标准”GC-MS和正交分离技术SFC-MS/MS。探讨了气相色谱检测的不同衍生化程序，并对多种衍生物的形成进行了描述和确认。综上所述，所调查的衍生化程序均未提供可接受的结果，以供进一步的方法开发以满足本项目的要求。SFC具有独特的选择性，能够克服这些问题，并以适当的选择性区分所有选定的类固醇，包括（前）孕激素、雄激素、皮质激素、雌激素和类固醇硫酸盐。特别是在对映体分析物的分离中，SFC已经显示出其作为GC替代品的潜力。在不同的色谱柱上成功分离了51种类固醇和类固醇硫酸盐，证明了SFC在内源性类固醇谱分析中的潜力。(C) 2018 Elsevier B.V.版权所有
In various fields of endocrinology, the determination of steroid hormones synthesised by the human body plays an important role. Research on central neurosteroids has been intensified within the last years, as they are discussed as biomarkers for various cognitive disorders. Their concentrations in cerebrospinal fluid (CSF) are considered to be regulated independently from peripheral fluids. For that reason, the challenging matrix CSF becomes a very interesting specimen for analysis. Concentrations are expected to be very low and available amount of CSF is limited. Thus, a comprehensive method for very sensitive quantification of a set of analytes as large as possible in one analytical aliquot is desired.However, high structural similarities of the selected panel of 51 steroids and steroid sulfates, including numerous isomers, challenges achievement of chromatographic selectivity.Since decades the analysis of endogenous steroids in various body fluids is mainly performed by gas chromatography (GC) coupled to (tandem) mass spectrometry (MS(/MS)). Due to the structure of the steroids of interest, derivatisation is performed to meet the analytical requirements for GC-MS(/MS). Most of the laboratories use a two-step derivatisation in multi-analyte assays that was already published in the 1980s. However, for some steroids this elaborate procedure yields multiple isomeric derivatives. Thus, some laboratories utilize (ultra) high performance liquid chromatography ((U)HPLC)-MS/MS as alternative but, even UHPLC is not able to separate some of the isomeric pairs. Supercritical fluid chromatography (SFC) as an orthogonal separation technique to GC and (U)HPLC may help to overcome these issues.Within this project the two most promising methods for endogenous steroid profiling were investigated and compared: the "gold standard" GC-MS and the orthogonal separation technique SFC-MS/MS. Different derivatisation procedures for gas chromatographic detection were explored and the formation of multiple derivatives described and confirmed. Taken together, none of the investigated derivatisation procedures provided acceptable results for further method development to meet the requirements of this project. SFC with its unique selectivity was able to overcome these issues and to distinguish all selected steroids, including (pro-)gestagens, androgens, corticoids, estrogens, and steroid sulfates with appropriate selectivity. Valued especially in the separation of enantiomeric analytes, SFC has shown its potential as alternative to GC. The successful separation of 51 steroids and steroid sulfates on different columns is presented to demonstrate the potential of SFC in endogenous steroid profiling. (C) 2018 Elsevier B.V. All rights reserved.