Page 20 - Analytical method for food addtives

E110: Sunset yellow 5

Analysis of beverages 3

Sample preparation
The samples were prepared as follows:
1 Quantitative determination by direct preparation using calibration graphs:
5 mL of the sample was transferred to a 25 mL flask and diluted with deionised
water to the mark.
2 Quantitative determination by standard addition: to 5 mL of the beverage
sample were added different amounts (2, 4, 6, 8 mg/L) of the dye to determine
and proceed as before.

Analysis of beverages
The samples were filtered through a Millipore filter before being injected into the
chromatographic system and all the experiments were carried out in duplicate.

HPLC conditions
Column Nova-Pak C18
Mobile phase Eluent A Methanol
Eluent B NaH PO /Na HPO buffer solution 0.1 M
2 4 2 4
pH=7
Gradient profile t (initial) 20 % eluent A, 80 % eluent B
0
t (2 min) 100 % eluent A
1
t (4 min) 100 % eluent A
2
t (5 min) 20 % eluent A, 80 % eluent B
0
Flow rate 2 mL/min
Injection volume 20 µL
Detection 520 nm