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2-(5-Benzoacridine)ethyl-p-toluenesulfonate as sensitive reagent for the determination of bile acids by HPLC with fluorescence detection and online atmospheric chemical ionization-mass spectrometric identification

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Abstract

2-(5-Benzoacridine)ethyl-p-toluenesulfonate (BAETS), a dual-sensitive probe, was reacted with bile acids in the presence of K2CO3 catalyst in dimethyl sulfoxide (DMSO) solvent to give BAETS–bile acid derivatives. Derivatives exhibited intense fluorescence (FL) with an excitation maximum at λ ex 270 nm and an emission maximum at λ em 510 nm. MS analysis using APCI-MS indicated that derivatives had excellent APCI-MS ionizability with percentage ionization δ values changing from 0 to 88.83% in aqueous acetonitrile and from 0 to 89.15% in aqueous methanol. The collision induced dissociation spectra of m/z [M + H]+ contained specific fragment ions at m/z [M + H−H2O]+, [M + H−2H2O]+, [M + H−3H2O]+, 347.3, and 290.1. Repeatability was good for LC separation of BAETS–bile acid derivatives with aqueous acetonitrile as mobile phase. The relative standard deviations (RSDs) of retention time and peak area at 6.6 nmol mL−1 levels with fluorescence detection (FL) were from 0.045 to 0.072% and from 2.16 to 2.73%, respectively. Excellent linear responses were observed, with regression coefficients >0.9995. The FL detection limits (S/N = 3) were in the range of 18.0–36.1 fmol. The online APCI-MS detection limits are in the range of 500–790 fmol (at a signal-to-noise ratio of 3).

Schematic diagram of derivatization procedure

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Acknowledgment

This work was supported by the National Science Foundation of China (No. 20075016) and supported by the 100 Talents Program of The Chinese Academy of Sciences (No. 328).

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Authors and Affiliations

  1. Northwest Plateau Institute of Biology, Chinese Academy of Sciences, Xining, 810001, China

    Jinmao You, Yanyan Fu, Zhiwei Sun & Yourui Suo

  2. The Key Laboratory of Life-Organic Analysis, College of Chemistry Science, Qufu Normal University, Qufu Shandong, 273165, China

    Jinmao You

  3. Graduate University of the Chinese Academy of Science, Beijing, 100049, China

    Zhiwei Sun

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  1. Jinmao You
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Correspondence to Jinmao You.

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You, J., Fu, Y., Sun, Z. et al. 2-(5-Benzoacridine)ethyl-p-toluenesulfonate as sensitive reagent for the determination of bile acids by HPLC with fluorescence detection and online atmospheric chemical ionization-mass spectrometric identification. Anal Bioanal Chem 396, 2657–2666 (2010). https://doi.org/10.1007/s00216-010-3467-4

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  • DOI: https://doi.org/10.1007/s00216-010-3467-4

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