Normal-phase high-performance counter-current chromatography for the fractionation of dissolved organic matter from a freshwater source.
Normal-phase high-performance counter-current chromatography (HPCCC) is used to obtain a preliminary fractionation of components in dissolved organic matter (DOM) from a freshwater source. The HPCCC solvent system involved a normal-phase approach with water/methanol (1:1) as the lower stationary phase and hexane/ethyl acetate (1:1) as the upper mobile phase. The critical experiment parameters were optimised: revolution speed 1800 rpm and flow rate 0.15 mL/min. Under these conditions 50 μL of a 0.50 mg/mL DOM solution was loaded. The detection wavelength was monitored at 330 nm in order to isolate the main portion of DOM, which includes substances such as carboxyl-rich alicyclic molecules. By optimising this system it was possible to isolate materials that, according to GC-MS, can be related to molecules with an analogous structural background. Where fraction analysis was not suitable for GC-MS, RP-HPLC with UV absorbance detection was used, showing unique chromatograms for each fraction at both 210 and 330 nm.
Irish Separation Science Cluster, National Centre for Sensor Research, School of Chemical Sciences, Dublin City University, Dublin, Ireland.
Sara
S
Sandron
Pavel N
PN
Nesterenko
Margaret V
MV
McCaul
Brian
B
Kelleher
Brett
B
Paull
D003377:Countercurrent Distribution / Q000295:instrumentation / Q000379:methods*; D005618:Fresh Water / Q000737:chemistry*; D009930:Organic Chemicals / Q000737:chemistry / Q000302:isolation & purification*; D012995:Solubility; D014874:Water Pollutants, Chemical / Q000737:chemistry / Q000302:isolation & purification*
D009930:Organic Chemicals; D014874:Water Pollutants, Chemical
Counter-current chromatography; Dissolved organic matter; Freshwater source; Sample fractionation