Analysis of carbohydrates by anion exchange chromatography and mass spectrometry.
A versatile liquid chromatographic platform has been developed for analysing underivatized carbohydrates using high performance anion exchange chromatography (HPAEC) followed by an inert PEEK splitter that splits the effluent to the integrated pulsed amperometric detector (IPAD) and to an on-line single quadrupole mass spectrometer (MS). Common eluents for HPAEC such as sodium hydroxide and sodium acetate are beneficial for the amperometric detection but not compatible with electrospray ionisation (ESI). Therefore a membrane-desalting device was installed after the splitter and prior to the ESI interface converting sodium hydroxide into water and sodium acetate into acetic acid. To enhance the sensitivity for the MS detection, 0.5 mmol/l lithium chloride was added after the membrane desalter to form lithium adducts of the carbohydrates. To compare sensitivity of IPAD and MS detection glucose, fructose, and sucrose were used as analytes. A calibration with external standards from 2.5 to 1000 pmole was performed showing a linear range over three orders of magnitude. Minimum detection limits (MDL) with IPAD were determined at 5 pmole levels for glucose to be 0.12 pmole, fructose 0.22 pmole and sucrose 0.11 pmole. With MS detection in the selected ion mode (SIM) the lithium adducts of the carbohydrates were detected obtaining MDL's for glucose of 1.49 pmole, fructose 1.19 pmole, and sucrose 0.36 pmole showing that under these conditions IPAD is 3-10 times more sensitive for those carbohydrates. The applicability of the method was demonstrated analysing carbohydrates in real world samples such as chicory inulin where polyfructans up to a molecular mass of 7000 g/mol were detected as quadrupoly charged lithium adducts. Furthermore mono-, di-, tri-, and oligosaccharides were detected in chicory coffee, honey and beer samples.
Bruggink
Cees
C
Dionex B.V., Lange Bunder 5, 4854 MB Bavel, The Netherlands. cees.bruggink@dionex.nl
Maurer
Rolf
R
Herrmann
Heiko
H
Cavalli
Silvano
S
Hoefler
Frank
F
D000837:Anion Exchange Resins; D001515:Beer / Q000032:analysis; D002138:Calibration; D002241:Carbohydrates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D003069:Coffee / Q000737:chemistry; D005632:Fructose / Q000032:analysis; D005947:Glucose / Q000032:analysis; D006722:Honey / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*; D013395:Sucrose / Q000032:analysis
D000837:Anion Exchange Resins; D002241:Carbohydrates; D003069:Coffee; D005632:Fructose; D013395:Sucrose; D005947:Glucose