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Combination of two spacers, here, 2\u2010amino\u20102\u2010hydroxymethyl\u2010propane\u20101,3\u2010diol and histidine, led to enhanced resolution of two model analytes (thiamine and pyridoxine) in model and real pharmaceutical samples (tablet drug dosage form). The addition of the spacers at relatively high concentration level (i.e., 20\u00a0\u00b5g\/mL) caused no significant interference and suppression of the analytical signal in the mass spectrometry detection step. Eco\u2010friendliness, practicality, and performance of the method were confirmed using the metric tools of the \u201cWhite Analytical Chemistry.\u201d High repeatability of the separation process could broaden the application possibilities of the hydrodynamically closed capillary electrophoresis system.<\/jats:p>","DOI":"10.1002\/elps.70143","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T06:41:31Z","timestamp":1787035291000},"page":"957-963","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Spacer\u2010Enhanced Resolution of Thiamine and Pyridoxine in Hydrodynamically Closed Two\u2010Dimensional Capillary Isotachophoresis Coupled With Tandem Mass Spectrometry"],"prefix":"10.1002","volume":"47","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-0642-3387","authenticated-orcid":false,"given":"Paula","family":"Cermakova","sequence":"first","affiliation":[{"name":"Department of 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