Documentation

Method Abstract 124

MA 124 / SULFONAMIDE ARTIFICIAL SWEETENERS IN FOOD PRODUCTS

ANALYSIS OF ACESULFAME, SACCHARIN AND CYCLAMATE BY HPLC WITH POST-COLUMN DERIVATIZATION

Nonnutritive sweeteners are widely used in foods and beverages. Since some studies have raised questions about long-term safety of these compounds, their concentration in food products is regulated. Among the artificial sweeteners used worldwide, Saccharin and Cyclamate cause most controversy and their use is restricted in many countries, including the US. Liquid chromatography is a method of choice for most artificial sweeteners, but Cyclamate analysis is complicated by the fact that this compound does not exhibit noticeable UV absorbance.

This method allows for simultaneous analysis of Acesulfame, Saccharin and Cyclamate by LC with post-column derivatization followed by fluorescence detection. Since the sweeteners are commonly used in combination, a single method of analysis is preferred.
 

MA 124METHOD

Analytical Conditions

Analytical Column: Reversed-phase column, C18, 2×100 mm
Temperature: 50 ˚C
Flow Rate: 0.2 mL/min
Mobile Phase: 0.02 M Potassium Phosphate Monobasic
       in water.  Isocratic for 8 min, followed by
       washing the column with 70 % ACN –
        30 % 0.02 M KH2PO4

Post-column Conditions

Post-column System: Pinnacle PCX
Reagent: Hexadecyltrimethyl ammonium bromide (10-3 M), 1,6 Diphenyl-1,3,5-hexatriene (4×10-6 M)
Reactor: 40 ˚C, 0.1 mL
Flow Rate: 0.3 mL/min
Detection: Fluorescence, λex = 365 nm, λem = 460 nm
  

REFERENCES
James F. Lawrence, Analyst, vol. 112, No 6 (1987), 879-881
James F. Lawrence, Claudette F. Charbonneau, J. Assoc. Anal. Chem., vol. 71, No 5 (1988), 934-937