Complete chromatographic separation of steroids, including 17α and 17β-estradiols, using a carbazole-based polymeric organic phase in both reversed and normal-phase HPLC
Complete chromatographic separation of steroids, including 17α and 17β-estradiols, using a carbazole-based polymeric organic phase in both reversed and normal-phase HPLC
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DOI：
10.1007/s00216-010-3601-3
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发表时间：
2010-05-01
影响因子：
4.3
通讯作者：
Ihara, Hirotaka
中科院分区：
文献类型：
--
作者：
Mallik, Abul K.;Shingo, Kaori;Ihara, Hirotaka
制备了末端带有三甲氧基硅基的聚（2-N-咔唑乙基丙烯酸酯）作为有机相，并将其固定在硅胶上。通过使用各种雌激素类固醇和皮质激素在反相和正相液相色谱中的保留行为的填充与这种咔唑基聚合物固定化硅胶（Sil-CEA）进行了研究。结果表明，Sil-CEA对8种甾体化合物的分离效果良好。使用Sil-CEA的最特异性分离可以通过高分离因子（例如，α =1.39，在甲醇-水（7：3，v/v）中，35 ℃），对于17 α和17 β-雌二醇，色谱分离中最困难的异构体对之一，而对于两种市售聚合ODS柱，在相同条件下，仅α =1.01。由于即使在正相移动的相中，如己烷-2-丙醇中，Sil-CEA也能保持优异的分离和保留顺序，因此估计CEA相通过比ODS预期的疏水效应更强的相互作用，如偶极-偶极、羰基-π和氢键相互作用，具有多种相互作用机制。
Poly(2-N-carbazolylethyl acrylate) with terminal trimethoxysilyl groups was prepared as an organic phase and immobilized onto silica. The retention behavior of the column packed with this carbazole-based polymer-immobilized silica (Sil-CEA) was investigated by using various estrogenic steroids and corticoids in both reversed-phase and normal-phase liquid chromatography. As a result, complete separation was confirmed for eight kinds of steroids with Sil-CEA. The most specific separation with Sil-CEA can be emphasized by the high separation factor (e.g., alpha=1.39 in methanol-water (7:3, v/v) at 35 degrees C) for 17 alpha and 17 beta-estradiols, one of the most difficult pairs of isomers in chromatographic separation, whereas for two kinds of commercially available polymeric ODS columns as references alpha=1.01, only, under the same conditions. Because the excellent separation and retention order with Sil-CEA was maintained even in a normal-phase mobile phase such as a hexane-2-propanol, it is estimated that the CEA phase has multiple interaction mechanisms through stronger interactions such as dipole-dipole, carbonyl-pi, and hydrogen bonding interactions than the hydrophobic effect expected with ODS.