Optimization and validation of a derivatization method for analysis of biogenic amines in wines using RP-HPLC-DAD

Abstract

An analytical method for determination of the biogenic amines: tryptamine, putrescine histamine, phenylethylamine, tyramine, cadaverine, spermidine and spermine in red and white wines using pre-column derivatization with dansyl chloride (DnsCl) and reversed phase-high performance liquid chromatography with diode array detector (RP-HPLC-DAD) was developed and optimized. Concentration of DnsCl, pH, time of reaction and evaporation under nitrogen were tested and optimized. Best recoveries, ranging from 72 to 102 % were obtained for all analyzed amines applying derivatization with 10 mg/mL of DnsCl for 1 h at 60 oC and pH 9. The developed method showed excellent linearity with R2>0.99 for all tested amines. The accuracy was checked with the standard addition method showing satisfactory recovery, good repeatability and reproducibility. The optimized and validated method was applied for analysis of white and red wines from Macedonian origin, observing higher total amounts of biogenic amines in red wines.

Keywords

• wine
• biogenic amines
• hplc
• derivatization
• dansyl chloride
• validation