Page 14 - Application Notebook - Solution for Food Development

LAAN-A-LM-E092

Application Liquid Chromatography Mass Spectrometry

News Simultaneous Analysis of Nine Sweeteners Using

Triple Quadrupole LC/MS/MS (LCMS-8040)
No.C121

Artificial sweeteners such as saccharin sodium, n Analysis of a Standard Mixture
aspartame, sucralose and acesulfame potassium fall Fig. 1 shows chromatograms measured from a 5 µL
under the category of specified additives in Japan's injected sample of a 10 ng/mL standard mixture of nine
Food Sanitation Act, for which each specified criteria sweeteners, analyzed with the analytical conditions
exist for their use in terms of eligible foods and shown in Table 1. Chromatograms at around the lower
amounts used. limit of quantitation (LLOQ) are shown in Fig. 2. The
Cyclamate, an artificial sweetener used in some regions retention time, calibration curve range, and correlation
of the world outside Japan, is an unspecified additive coefficient for each compound are shown in Table 2.
within Japan, for which inspection is required on A calibration point accuracy of within 100 ± 20 % and
specific imported foods. a percentage of area repeatability (%RSD) of within
In light of these situations, there is a demand for 20 % were employed. Good linearity was obtained for
analyses of various different sweeteners, not only the all compounds with a correlation coefficient of 0.997 or
quantitative testing of permitted sweeteners but also higher.
the testing of unspecified sweetener additives.
This article presents a simultaneous analysis of nine
sweeteners including both specified additives and
unspecified additives, using the LCMS-8040 high-
performance liquid chromatograph-triple quadrupole
mass spectrometer.

(×10,000) (×100,000)
Acesulfame potassium 2.5 Dulcin
2.5
0.0 0.0
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min
(×1,000) (×100,000)
2.0 Saccharin 2.5 Neotame
1.0
0.0 0.0
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min
(×10,000) (×100)
1.0 Cyclamate Rebaudioside A
0.5 5.0
0.0 0.0
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min
(×1,000) (×100)
5.0 Sucralose 5.0 Stevioside
0.0 0.0
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min
(×10,000)
1.0 Aspartame
0.5
0.0
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 min
Fig. 1 Chromatograms from a 10 ng/mL Standard Mixture of Nine Sweeteners

Table 1 Analytical Conditions
Column : Unison UK-C18 (150 mm L. × 3.0 mm I.D., 3.0 µm)
Mobile Phases : A 5 mmol/L Ammonium Formate - Water
: B Methanol
Gradient : B Conc. 0 % (0.0 - 2.0 min) → 70 % (4.5 min) → 90 % (8.0 - 12.0 min) → 0 % (12.01-15.0 min)
Flowrate : 0.2 mL/min
Column Temperature : 40 ˚C
Injection Volume : 5 µL
Probe Voltage : ＋ 4.5 kV (ESI-positive mode) / -3.5 kV (ESI-negative mode)
DL Temperature : 300 ˚C
Block Heater Temperature : 500 ˚C
Nebulizing Gas Flow : 3 L/min
Drying Gas Flow : 15 L/min