Highly sensitive UHPLC—MS/MS method for quantification of ethylenediamine-N,N′-di-2-(3-cyclohexyl) propanoic acid derivatives in mouse serum

Acta Chromatographicae

Biljana K. Tubić¹  Miroslav M. Savić²  Bojan D. Marković²  Sandra S. Vladimirov²  Branka M. Ivković²  Slavica M. Ristić³  Tibor J. Sabo⁴  Jelena M. Poljarević⁴ 

[1] 1Agency for Medicines and Medical Devices of Bosnia and Herzegovina, St. Veljka Mlađenovića bb, 78000 Banjaluka, Bosnia and Herzegovina;2Faculty of Pharmacy, University of Belgrade, St. Vojvode Stepe 450, 11040 Belgrade, Republic of Serbia;3Galenika a.d., Institute for Research and Development, St. Pasterova 2, 11000 Belgrade, Republic of Serbia;4Faculty of Chemistry, University of Belgrade, St. Studentski trg 12-16, 11000 Belgrade, Republic of Serbia

关键词: UHPLC—MS/MS;    protein precipitation sample preparation;    (S;    S)-ethylenediamine-N;    N′-di-2-(3-cyclohexyl) propanoic acid esters;    cytotoxic activity;    metabolite;   

DOI  :  10.1556/1326.2017.29.2.7

学科分类：化学（综合）

来源: Akademiai Kiado Rt.

【 摘 要 】

A series of new (S,S)-ethylenediamine-N,N′-di-2-(3-cyclohexyl)propanoate esters has shown cytotoxic activity towards human leukemic cell lines. The aim of this study was to develop and validate a bioanalytical method for quantification of (S,S)-O,O-diethyl-ethylenediamine-N,N′-di-2-(3-cyclohexyl)propanoate dihydrochlorides (DE-EDCP) and its metabolite, substituted propanoic acid (EDCP), in mouse serum by ultra high-performance liquid chromatography—tandem mass spectrometry (UHPLC—MS/MS). Structural analog, derivative of 1,3-propanediamine, was used as an internal standard (IS). Sample preparation employed protein precipitation by acetonitrile and subsequent centrifugation. Optimal UHPLC separation conditions were set to achieve simultaneous determination of both compounds in a short run time of 6 min. Additionally, the selected reaction monitoring (SRM) mode developed in this method allowed a highly sensitive, accurate, and precise identification of compounds of interest. The lower limit of quantitation (LOQ) was 1.3 ng mL−1 for DE-EDCP and 0.3 μg mL−1 for EDCP. The calibration curves were linear over the concentration range of 1.3–26.7 ng mL−1 and 0.3–6.7 μg mL−1 for DE-EDCP and EDCP, respectively. Precision (%CV) and accuracy (%RE) for DE-EDCP and EDCP ranged from 3.5% to 16.0% and from 1.8% to 14.4%, respectively.

【 授权许可】

Unknown