title: Micro-scale strategy to detect spermine and spermidine by MALDI-TOF MS in foods and identification of apoptosis-related proteins by nano-flow UPLC-MS/MS after treatment with spermine and spermidine.
abstract: Spermine and spermidine are multiple-nitrogen compounds found in many foods. Both compounds are essential for cell growth and human health. This study established a simple and fast method of detecting spermine and spermidine in food samples by matrix-assisted laser desorption/ionization combined with time-of-flight mass spectrometry (MALDI-TOF MS). After a simple sample preparation procedure, spermine and spermidine were directly detected by MALDI-TOF MS with no additional purification procedure. The calibration curves for spermine and spermidine ranged from 0.1 to 10 μg/mL. In intra- and inter-batch assays of three different concentrations of spermine and spermidine, all relative standard deviations and relative errors were below 18.9%. These experimental results confirmed the practicability and effectiveness of the proposed MALDI-TOF MS method for fast determination of spermine and spermidine in food samples. Furthermore, since spermine and spermidine have important roles in apoptosis, up-regulation and down-regulation of spermine and spermidine during apoptosis were analyzed. After treating NRK-52E cells with spermine and spermidine, the cells were lysed, and cell proteins were collected, and digested. Apoptosis-related proteins were then identified by tandem MS.
authors.affiliation: Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Huai-Hsin
authors.initials: HH
authors.lastname: Su
authors.affiliation: Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Lea-Yea
authors.initials: LY
authors.lastname: Chuang
authors.affiliation: Department of Chemistry, College of Science, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Wei-Lung
authors.initials: WL
authors.lastname: Tseng
authors.affiliation: Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; Research Center for Environmental Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; Institute of Medical Science and Technology, National Sun Yat-sen University, Kaohsiung 80424, Taiwan. Electronic address: cylu@kmu.edu.tw.
authors.forename: Chi-Yu
authors.initials: CY
authors.lastname: Lu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000032:analysis / Q000378:metabolism*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005504:Food Analysis / Q000379:methods*; D057230:Limit of Detection; D051381:Rats; D015203:Reproducibility of Results; D013095:Spermidine / Q000032:analysis* / Q000494:pharmacology; D013096:Spermine / Q000032:analysis* / Q000494:pharmacology; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D051017:Apoptosis Regulatory Proteins; D013096:Spermine; D013095:Spermidine
keywords: Apoptosis; Food; MALDI–TOF MS; NanoUPLC–MS/MS; Spermidine; Spermine