4 May / June 2016
Profiling and Quantification of Mono and Disaccharides and Selected Alditols in Juice, Beer, Wine, and Whiskey Using UHPLC with Mass Detection
by Mark E. Benvenuti, Gareth Cleland, and Jennifer Burgess Waters Corporation, Milford, MA, USA
Detector (mass detection) coupled to the ACQUITY Arc System (UHPLC) for the profiling and quantification of sugars in juice, wine, beer, and whiskey samples.
EXPERIMENTAL LC conditions LC system:
Data system: Runtime:
Column: XBridge® UPLC
Empower® 40.0 min
ACQUITY Arc 3
XP BEH Amide 2.5 µm, 3.0 x 150 mm
Column temp.: 85°C Mobile phase A: 90% acetonitrile: 5% IPA:5% water* Mobile phase B: 80% acetonitrile: 20% water* Flow rate:
0.8 mL/min Injection volume: 1 µL SIR channels:
MS conditions MS system:
Analyte ACQUITY QDa (Performance mode)
Ionisation mode: ESI capillary voltage:
Cone voltage: Probe temp:
0.8 V 5.0 V
600°C
Acquisition rate: 2 Hz Full scan: Curve fit:
Smoothing: 50 to 800 Hz
Quadratic, 1/x weighting Mean filter, Level 7
Arabinose Fructose Glucose Inositol Sorbitol Mannitol Sucrose Maltose
Maltotriose
Formula C5
C6 C6 C6 C6 C6
H10 H12 H12 H12 H14 H14
C12 C12 C18
O5 O6 O6 O6 O6 O6
H22 H22 H32
O11 O11 O16
SIR (m/z) ([M+Cl]- 185 215 215 215 217 217 377 377 539
ion)
gradient: 1.
2. 3. 4. 5. 6.
Time (min)
Initial 4.5
18.0 25.0 25.1 40.0
Flow rate (mL/min)
0.8 0.8 0.8 0.8 0.8 0.8
Sugars and sugar alcohols are classes of carbohydrates that are important in human nutrition and natural constituents of foods. With the increasing incidence of obesity and diabetes across the developed world, interest in monitoring sugar intake has vastly increased in recent years. Consequently, there are now requirements to provide accurate information on product labelling in order to comply with increasingly stringent regulatory demands. Profiling the sugar content of products is also a useful tool in assessing product authenticity and potential adulteration. The analysis of sugars and sugar alcohols remains a challenging application, owing to the lack of chromophores and the similarity between these molecules - many of these sugar compounds are isomers of one another. Due to its separation power, accuracy, and speed of analysis, HPLC has become the method of choice for the analysis of sugars. An alternative to RI and ELS detection is the use of mass detection with electrospray ionisation (ESI). Mass detection is complementary to traditional detectors used for LC. It offers the opportunity to decrease detection limits and also to obtain mass spectral information on the components in the sample. The combination of both chromatographic retention time and mass information results in increased selectivity for the analysis of sugars and sugar alcohols. Here we show the application of the Waters® QDa®
ACQUITY®
%A 100
100 0 0
100 100
*Both containing 500 ppb guanidine hydrochloride and 0.05% diethylamine.
%B 0
0
100 100 0 0
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