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Molecules 2017, 22(7), 1198; doi:10.3390/molecules22071198

Application of An Improved HPLC-FL Method to Screen Serine Palmitoyl Transferase Inhibitors

1
Dipartimento di Farmacia, Università di Pisa, Via Bonanno 6, 56126-Pisa, Italy
2
Laboratorio di Biochimica e Biologia Molecolare, Dipartimento di Scienze della Salute, Via A. di Rudinì 8, 20142 Milano, Italy
*
Author to whom correspondence should be addressed.
Received: 5 July 2017 / Revised: 14 July 2017 / Accepted: 14 July 2017 / Published: 17 July 2017
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Abstract

In this work, we reported the application and validation of an improved high-performance liquid chromatography method coupled with a fluorimetric detector (HPLC-FL) to screen the activity of two heterocyclic derivatives reported as serine palmitoyl transferase (SPT) inhibitors. The analytical conditions were optimized in terms of the derivatization procedure, chromatographic condition, extraction procedure, and method validation according to EMEA guidelines. Once fully optimized, the method was applied to assess the SPT-inhibitory activity of the above-mentioned derivatives and of the reference inhibitor myriocin. The obtained results, expressed as a percentage of residual SPT activity, were compared to those obtained with the reference radio immune assay (RIA). The good correlation between the two types of assay demonstrated that the improved HPLC-FL method is suitable for a preliminary and rapid screening of potential SPT-inhibitors. View Full-Text
Keywords: HPLC; serine palmitoyl transferase; SPT; screening; enzymatic assay HPLC; serine palmitoyl transferase; SPT; screening; enzymatic assay
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Bertini, S.; Saccomanni, G.; Carlo, S.D.; Digiacomo, M.; Gargini, C.; Piano, I.; Campisi, G.M.; Ghidoni, R.; Macchia, M.; Manera, C. Application of An Improved HPLC-FL Method to Screen Serine Palmitoyl Transferase Inhibitors. Molecules 2017, 22, 1198.

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