Simultaneous determination nucleosides in marine organisms using ultrasound‐assisted extraction followed by hydrophilic interaction liquid chromatography–electrospray ionization time‐of‐flight mass spectrometry

Abstract

A new method has been developed based on ultrasound-assisted extraction (UAE) followed by hydrophilic interaction chromatography (HILIC) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF/MS) for the simultaneous determination of 16 nucleosides and nucleobases in medicinal extracts of various marine organisms. The separation was achieved on a Venusil HILIC column (250×4.6 mm id, 5 μm) and gradient elution using a solution of acetonitrile and buffer (0.20% formic acid and 20 mmol/L ammonium acetate) as the mobile phase. Identification of the 16 target nucleosides and nucleobases was based on the retention time, UV spectra, and mass measurements of the protonated molecules ([M+H](+)) and main fragment ions (ESI-TOF/MS). In addition, non-target compounds of 2'-deoxyinosine and four other amino acids were also tentatively identified by ESI-TOF/MS. The 16 target compounds were quantified by HILIC-ESI-TOF/MS under optimized mass conditions. All calibration curves showed good linearity (r(2)>0.9951). The recoveries were 84.72-124.10%, and the limits of detection of the 16 target compounds were 0.6-130.0 ng/mL. The developed method was applied to quantify the target compounds in 15 batches of various marine organisms. The method has potential applicability for the identification and determination of highly polar and low-concentration active compounds in marine organisms.

Chemometrics for comprehensive analysis of nucleobases, nucleosides, and nucleotides in Siraitiae Fructus by hydrophilic interaction ultra high performance liquid chromatography coupled with triple‐quadrupole linear ion‐trap tandem mass spectrometry

“…Many techniques were used for analysis of nucleobases, nucleosides, and nucleotides including CE , RP‐LC , and HILIC combined with UV or MS detection . Among these methods, HILIC coupled to triple quadrupole tandem mass spectrometry (QqQ‐MS 2 ) was often regarded as the proper method to analyze these compounds with high polarity and similar structure.…”

Characterization of Nucleosides and Nucleobases in Natural Cordyceps by HILIC–ESI/TOF/MS and HILIC–ESI/MS

“…In contrast to RP–HPLC, a hydrophilic polar stationary phase was used in HILIC, showing good reservation and separation efficiency for polar compounds. HILIC has been used in the analysis of several polar compounds in TCMs such as Ligusticum chuanxiong [26] and several medicinal marine organisms [27]. However, the possibility of simultaneously separating the 15 nucleosides, nucleobases and other components in the Cordyceps samples by using HILIC remains unknown.…”

Evaluation of Brewer’s Spent Yeast To Produce Flavor Enhancer Nucleotides: Influence of Serial Repitching

“…Other techniques include enzymatic assay, capillary electrophoresis, capillary electrophoresis–inductively coupled plasma mass spectrometry, capillary electrochromatography, and ion-pair reversed-phase chromatography . More recently, hydrophilic interaction liquid chromatography (HILIC) has found increased use for the separation of highly polar molecules, namely, nucleotides that are often unretained under RP-HPLC. − …”