SUCRALOSE ANALYSIS USING EVAPORATIVE LIGHT SCATTERING DETECTION (ELSD)
Mark E. Benvenuti, and Joseph P. Romano
Waters Corporation, Milford, MA 01757
Setup for Waters 2420 ELSD Detector
Gas (N2)
45.0 psi
Nebulizer 60%
Drift Tube
1 Temperature 43°C
1
212.3 ppm
0.50
1.50
2.50
3.50
4.50
5.50
6.50
10
400
600
800
Diet Fudge Soft Drink
1000
500 Light Scattering
1
2
3
4
2
Sucralose
Fructose
Glucose
Sucrose
1071ppm
1165 ppm
1305 ppm
1278 ppm
4
1
1 Sucralose 198.6 ppm
3
0.50
2.00
6.00
10.0
Minutes
14.0
500 Light Scatter-
2.50
3.50
Minutes
4.50
5.50
1
Diet Ice Tea
% RSD = 0.03 %
% RSD = 2.32
3.5
1 Sucralose 178.8 ppm
0.50
4.0
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4.5
4.5
4.0
Minutes
2.00
6.0
10.00
14.0
18.0
Minutes
Conclusion:
Lamp
Minutes
Glucose (Dextrose)
6.50
Nebulizer
3.5
Sucralose
0.39% By Weight
18.0
Reproducibility: 7 injections of standard 1
Ret Time
Area
%
1.50
Unknown
Sucralose was obtained from Tate and Lyle, Decatur, Illinois
A 1 gram packet of non caloric sweetener containing
sucralose (actual wt 1.095 grams) was dissolved in
50 ml of 75% acetonitrile/ water and quantitated in
the same way as the diet beverages with the following
result.
1400 Light Scattering Units
200
ppm
Standard Preparation:
A sucralose standard was prepared by dissolving
0.208 grams of sucralose in 75% acetonitrile/ water
and diluting to 200 ml with same. This standard was
diluted 1:2, 1:5, 1:10, 1:20, 1:50, and 1:100 with
the same matrix to produce a seven point calibration
curve as shown.
r2= 0.9978
Several commercial diet soft drinks containing sucralose were purchase at a local grocery. Where appropriate, sonnication was used to remove carbonation followed by filtration through a 0.45 micron hydrophilic filter. This was the only sample
preparation necessary. One part sample and three
parts acetonitrile were mixed and injected into the
chromatographic system. The injection volume was 50
microliters.
Minutes
Chromatogram of Common Sugars and Sucralose
A mixed standard was prepared by dissolving 0.107
grams of sucralose, 0.116 grams fructose, 0.131
grams glucose, and 0.128 grams sucrose in 75% acetonitrile/ water, and diluting to 100 ml with same. The
sugars were reagent grade.
1 Sucralose
11 Light Scattering Units
Gain Setting
Diet Cola
10 Light Scattering Units
Another product gaining in popularity is sucralose.
Derived from sucrose through the substitution of three
chlorine
atoms, this compound is a saccharide and thus has no
significant UV activity. For this reason, we shall show
the efficacy of Evaporative Light Scattering Detection
( ELSD ) for the analysis of this compound.
Due to the non- linear response of ELSD data,
a quadratic fit is appropriate
25 Light Scattering Units
Waters Alliance™ Solvent and Sample Manager
Data System: Empower™
Eluent
75 % Acetonitrile, 25% Water
Column
YMC-Pack Polyamine II, 4.6 X 250 mm,
5 µm
Temp
35° C
Flow Rate 1.0 ml/min
Unknown
For health reasons, the use of non-nutritive sweeteners
in soft drink and other products has markedly increased in the past few years. Of these, compounds
such as Acesulfame and Aspartame are UV active and
can easily be detected using this technique.
Sample Preparation:
Linearity
12 Light Scattering Units
HPLC Conditions
5 X 10 6 Peak A rea
Introduction:
1.50
2.50
3.50
4.50
5.50
6.50
Evaporative Light Scattering can be used for the
quantitation of Sucralose. Sample preparation for
soft drink matrices is simple, requiring a filtration
step and decarbonation where appropiate. ELSD
analysis can also be used to confirm the absence of
sugars in a diet preparation.
Minutes
Lit Code Number 720001354EN
©2005 Waters Corporation