High performance liquid chromatographic determination of N- nitrosoamines by pre-column fluorescence derivatization with acridone-N- acetyl chloride

Jinmao You, Xinjun Fan, Wenjian Lao, Qingyu Ou, Qingcun Zhu

Research output: Contribution to journalArticle

24 Scopus citations


A sensitive high performance liquid chromatographic method for the determination of N-nitrosoamines with pre-column fluorescence derivatization has been developed. N-nitrosoamines are first changed into secondary amines using denitrosation reagent, then react with acridone-N-acetyl chloride (ARC- Cl) to produce corresponding secondary amine derivatives, which exhibit a strong fluorescence. Maximum emission for ARC derivatives is 430 nm (λ(ex) 404 nm). The labelled derivatives are very stable, less than 4% decomposition occurs after heating at 40°C for 24 h. Fluorescence intensities of derivatives are higher in neutral and alkaline than in acidic solutions. This method, in conjunction with a multi-gradient program, offers a baseline resolution of the ARC derivatives from a linear acetonitrile gradient. Separation is carried out on a reverse phase C18 column. Derivatization and chromatographic conditions are optimized. The relative standard deviation (n = 6) at an analytical concentration of 10 pmol of each N-nitroamine is less than 4.5%. The detection limits at the fmol level. The method described is also suitable for analysis of other amino compounds in different biological samples.

Original languageEnglish (US)
Pages (from-to)437-449
Number of pages13
Issue number2
Publication statusPublished - Jan 1 1999
Externally publishedYes



  • Acridone-N-acetyl chloride (ARC-Cl)
  • Column liquid chromatography
  • Denitrosation
  • Fluorescence derivatization
  • N-nitroso compounds

ASJC Scopus subject areas

  • Analytical Chemistry

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