Page 348 - Analytical method for food addtives

E954: Saccharin 235

comparing the absorption properties of the sample with those of the standard
substance after either recording the absorption curve or taking measurements at
different wavelengths in the relevant range for both sample and standard.
Laboratories equipped with fixed-wavelength detectors should carry out separate
runs for determination of cyclamate and saccharin at the wavelengths concerned.
This method allows the determination of sorbic acid at the same conditions as
chosen for saccharin (of wavelength 265 nm). Whenever the determination of
sorbic acid with improved sensitivity is required, an additional wavelength switch
to 260 nm is recommended.
Note 1. As sorbic acid is a late-eluting compound, the risk of interference in the
next run has to be taken into account.
Note 2. If the separating column and mobile phase (as stated in Table 26.1) are
used, it has been found satisfactory to adopt the following experimental conditions.
Flow 1.7 mL/min
UV detection 200 nm (sodium cyclamate)
265 nm (saccharin)
260 nm (sorbic acid)
Volume injected 20 µL

Determination by HPLC
To carry out the determination by the external standard method, integrate the peak
areas or determine the peak heights and compare the results with the corresponding
values for the standard substance with the nearest peak area/height, or use a
calibration curve. In the case of a calibration curve additional solutions with
concentrations within the linear range may be prepared for the calibration graph.
Inject equal volumes of the sample and standard test solutions. Check the
linearity of the calibration function.
(For further details see BS EN 1379:1997.) 12

Foodstuffs – determination of acesulphame-K, aspartame and saccharin –
high performance liquid chromatographic method 13
Scope
An HPLC method for the determination of acesulphame-K, aspartame and saccha-
rin. It also allows the determination of caffeine, sorbic acid and benzoic acid in
foodstuffs.

Principle
The sample is extracted or diluted with water. If necessary, the sample solution
with the intense sweeteners is purified on a solid-phase extraction column or with
Carrez reagents. The intense sweeteners in the sample test solution are separated
on an HPLC-reversed phase chromatography column and determined
spectrometrically at a wavelength of 220 nm.