Determination of polyamines in human saliva by high-performance liquid chromatography with fluorescence detection

J Chromatogr B Biomed Sci Appl. 2001 Jun 5;757(1):111-7. doi: 10.1016/s0378-4347(01)00130-x.

Abstract

A high-performance liquid chromatographic method for the determination of polyamines (spermine, spermidine and putrescine) in human saliva was developed. This method is based on pre-column derivatization with o-phthaldialdehyde (OPA). The derivatives were separated on a Nucleosil ODS column (250x4.6 mm I.D.; 5 microm). The gradient elution was performed with two mobile phases A (water) and B (methanol) at a flow rate of 0.8 ml/min. The column eluate was monitored by fluorescence detection (excitation, 360 nm; emission, 510 nm). The within- and between-assay coefficients of variation for all the compounds were below 5%. The detection limits for spermine, spermidine and putrescine were 0.04, 0.05 and 0.06 nmol/ml, respectively. The recovery was greater than 90%. Our analytical technique requires neither preliminary extraction with an organic solvent, nor long multi-step procedures. For saliva samples, this is a simple, rapid and highly reproducible method that can be easily applied to the routine determination of salivary polyamines, whose levels increase early in several pathological conditions.

MeSH terms

  • Biogenic Polyamines / analysis*
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Reproducibility of Results
  • Saliva / chemistry*
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence

Substances

  • Biogenic Polyamines