GOA L
To rapidly analyze artificial sweeteners in
beverages and canned fruits using SPE followed
by the ACQUITY UPLC® H-Class System
with ELSDs.
BAC KG ROU N D
The increase in obesity due to the consumption
of sugary foods has reached epidemic levels,
especially in the developed world. Food
manufacturers are therefore resorting to non-
caloric sweeteners to reverse this trend. Ideally,
a blend of sweeteners will mimic the taste
and sensation of sugar as closely as possible.
Common sweeteners used today are aspartame,
saccharin, acesulfame K, and sucralose. Less
common substances include sodium cyclamate,
NHDC (neohesperidin dihydrochalcone), and
neotame. Regulations vary as to the legality
of certain sweeteners in various parts of
the world.
Because of the different regulations, it is
desirable to have a rapid method that can
be used to look for the presence of these
sweeteners. Acesulfame K, saccharin, and
aspartame, along with caffeine and certain
preservatives, are routinely analyzed in soft
drinks using UV detection. However, cyclamate
and sucralose are UV transparent rendering UV
detection unfeasible.
Run multiple samples in under five minutes using the
ACQUITY UPLC H-Class System coupled with an ELSD.
Fast Screening for Non-Nutritive Sweeteners using the ACQUITY UPLC H-Class System with ELSD
Figure 1. ELSD Chromatogram of the UPLC separation of seven sweeteners.
Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60
1
76
5
4
3
2
30 L
SU
1- AcesulfameK- 57.6 ppm2- Na_Saccharin- 87.6 ppm3- Na_Cyclamate- 172.8 ppm4- Sucralose- 69.6 ppm5- Aspartame- 166.8 ppm6- NHDC- 58.8 ppm7- Neotame- 56.4 ppm
T H E SO LU T IO N
Using a Waters® ACQUITY UPLC H-Class System equipped with an
Evaporative Light Scattering Detector (ELSD), separation of the seven listed
analytes can be achieved in less than five minutes, as shown in Figure 1.
This method uses UPLC® HSS T3 Column chemistry and a ternary gradient.
A calibration curve using a log linear fit is employed for quantification. This
procedure can also be used for qualitative screening.
Waters Corporation34 Maple StreetMilford, MA 01757 U.S.A.T: 1 508 478 2000F: 1 508 872 1990www.waters.com
For simple beverage samples, filtration and, if
necessary, degassing to remove carbonation can be
employed prior to direct injection of the samples.
For more complicated matrices, a simple cleanup
using Sep-Pak® C18 Cartridge can be used for
sample preparation. This procedure is described in
the European Commission’s validated method for
the analysis of nine sweeteners (EUR 22727 EN,
A. Wasik and M. Buchgraber, 2007). The sample,
canned fruit in this example, was homogenized
and thoroughly mixed with a buffer solution, then
centrifuged. The supernatant was loaded onto a
Sep-Pak C18 Cartridge, washed, and the analytes
eluted with methanol. Chromatograms from a diet
soft drink and canned diet pears carried through
this procedure is shown in Figure 3. Three of the
sweeteners were identified and quantified in the soft
drink and two were identified and quantified in the
canned pear sample.SUMMA RY
Using the ACQUITY UPLC H-Class System with ELSD detection, baseline
separation of seven sweeteners can be achieved in less than five minutes. The
short run time allows many samples to be run either for screening purposes,
or as a quantitative QC check.
Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60
12
3
2
1
7 LS
U7 LS
U
1-AcesulfameK- 79 ppm2- Sucralose- 59 ppm3- Aspartame- 214 ppm
1-AcesulfameK- 42 ppm2- Sucralose- 6 ppm
A: Diet Soft Drink
B: Canned Diet Pears
Figure 3. ELSD chromatograms of two samples following SPE cleanup. A: diet soft drink; B: canned diet pears. The calculated concentrations of each of the detected sweeteners is shown (with sample preparation adjustments accounted for in the final value).
Figure 2. The ACQUITY UPLC H-Class System with ELSD detection.
Waters, ACQUITY UPLC, UPLC, and Sep-Pak are registered trademarks of Waters Corporation. The Science of What’s Possible is a trademark of Waters Corporation. All other trademarks are the property of their respective owners.
©2011 Waters Corporation. Produced in the U.S.A.November 2011 720004166EN AO-PDF
