Comparison of Strategies for the Determination of Sterol Sulfates via GC-MS Leading to a Novel Deconjugation-Derivatization Protocol
Comparison of Strategies for the Determination of Sterol Sulfates via GC-MS Leading to a Novel Deconjugation-Derivatization Protocol
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DOI：
10.3390/molecules24132353
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发表时间：
2019-07-01
期刊：
影响因子：
4.6
通讯作者：
Bracher, Franz
中科院分区：
文献类型：
--
作者：
Junker, Julia;Chong, Isabelle;Bracher, Franz
甾醇的磺基共轭物作为神经甾体、神经递质和离子通道配体在健康和疾病中发挥重要作用。在大多数情况下，甾醇共轭物分析是用液相色谱-质谱法进行的。这是一种有价值的常规分析工具，其优点是无需预先裂解和/或衍生化即可直接进行甾醇硫酸盐分析。气相色谱-质谱联用（GC-MS）是甾体组学领域的一种重要发现工具，但其分析受到强制性去共轭和衍生化的阻碍。尽管样品后处理困难，GC-MS是非靶向分析和类固醇分析不可或缺的工具。有没有一般的样品制备使用GC-MS分析硫酸甾醇的协议。在这项研究中，我们提出了一个重新调查和评价不同的去共轭和衍生化程序与一组代表性的硫酸甾醇。考察了三甲基硅基（TMS）、肟基-三甲基硅基（MO-TMS）和三氟乙酰基（TFA）衍生物的优缺点。不同公布的程序，包括酶和化学裂解，甾醇硫酸酯解共轭，重新调查和检查不同的甾醇硫酸酯。最后，我们提出了一种新的化学裂解甾醇硫酸酯的协议，允许同时去共轭和衍生化，简化基于GC-MS的甾醇硫酸酯分析。
Sulfoconjugates of sterols play important roles as neurosteroids, neurotransmitters, and ion channel ligands in health and disease. In most cases, sterol conjugate analysis is performed with liquid chromatography-mass spectrometry. This is a valuable tool for routine analytics with the advantage of direct sterol sulfates analysis without previous cleavage and/or derivatization. The complementary technique gas chromatography-mass spectrometry (GC-MS) is a preeminent discovery tool in the field of sterolomics, but the analysis of sterol sulfates is hampered by mandatory deconjugation and derivatization. Despite the difficulties in sample workup, GC-MS is an indispensable tool for untargeted analysis and steroid profiling. There are no general sample preparation protocols for sterol sulfate analysis using GC-MS. In this study we present a reinvestigation and evaluation of different deconjugation and derivatization procedures with a set of representative sterol sulfates. The advantages and disadvantages of trimethylsilyl (TMS), methyloxime-trimethylsilyl (MO-TMS), and trifluoroacetyl (TFA) derivatives were examined. Different published procedures of sterol sulfate deconjugation, including enzymatic and chemical cleavage, were reinvestigated and examined for diverse sterol sulfates. Finally, we present a new protocol for the chemical cleavage of sterol sulfates, allowing for simultaneous deconjugation and derivatization, simplifying GC-MS based sterol sulfate analysis.