High-Throughput LC–MS/MS Based Simultaneous Determination of Polyamines Including N-Acetylated Forms in Human Saliva and the Diagnostic Approach to Breast Cancer Patients

Abstract

The determination of polyamines and their N-acetylated forms was performed by ultraperformance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). The polyamines efficiently reacted with 4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (DBD-F) in 0.1 M borax (pH 9.3) at 60 °C for 30 min. The resulting derivatives were analyzed by electrospray ionization (ESI)-MS and sensitively detected by selected reaction monitoring (SRM). Furthermore, a rapid separation of the polyamine derivatives within 10 min was performed by UPLC using an antipressurized column packed with 1.7-μm octadecylsilyl (ODS) silica gel. The limits of detection (S/N = 3) on the SRM chromatograms were at the attomole level (9-43 amol). This procedure was used to successfully determine 11 polyamines, including their N-acetylated forms, in the saliva of patients with primary and relapsed breast cancer and healthy volunteers. The level of several polyamines (Ac-PUT, Ac-SPD, Ac-SPM, DAc-SPD, and DAc-SPM) increases in breast cancer patients. Furthermore, the levels of three polyamines (Ac-SPM, DAc-SPD, and DAc-SPM) were significantly higher only in the relapsed patients. The present method proved highly sensitive and is characterized by specificity and feasibility for sample analysis. Consequently, the proposed method is useful for the noninvasive salivary diagnosis of cancer patients and could be applied to determine polyamines in several specimens of biological nature.

Pharmacometabolomics study identifies circulating spermidine and tryptophan as potential biomarkers associated with the complete pathological response to trastuzumab-paclitaxel neoadjuvant therapy in HER-2 positive breast cancer

“…However, a reduced catabolic clearance of Spd could also explain the higher concentration of Spd observed. Further investigation into the clearance of acetyl derivatives of polyamine [33] may give a more comprehensive explanation for the elevated Spd serum levels observed in the GR group.…”

Derivatization-based High-throughput Bioanalysis by LC-MS

“…[18][19][20][21][22][23] The polyamines in the fingernail of lung-cancer patients were also determined after labeling with DBD-F and detected by LC-TOF-MS. 24 DBD-F was also applied to the determination of polyamines, including its N-acetylated form in the saliva of breast-cancer patients by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). 25 The increase and decrease of specific polyamines were identified in breast-cancer patients. The diagnosis of breast cancer using saliva is now possible by this method.…”

Diagnostic Approach to Disease Using Non-invasive Samples Based on Derivatization and LC-ESI-MS/MS

“…To solve this drawback, the derivatization method was adopted for the target analytes. 134,135) The resulting derivatives provide an efficient separation and/or high detection sensitivity due to increase in the hydrophobicity and positivity. The polyamines and their N-acetyl compounds in saliva were determined as an application.…”