CARBOHYDRATES
CE-LEDIF
275 nm
AN 2.003

Polysaccharides Analysis

9-Aminopyrene-1,4,6-trisulfonate (APTS) is a dye that is frequently used for the analysis of mono or oligosaccharides. The labeling of sugars involves a reductive amination of the reductive function of the mono or oligosaccharides followed by reaction with the dye.

APTS is routinely used in Capillary Electrophoresis separation. In this note, we analyze oligosaccharides labeled with APTS  with a 480nm LED.

CE-LEDIF Analysis

Instruments

Capillary electrophoresis: Agilent Technologies 7100 CE

Detector: Picometrics ZETALIF LED with 480nm/30nm LED

Sample

Dextran 5000 labeled with APTS

Labeling

500µg dextran 5000 + 15µL APTS solution (5mg in 75µL acetic acid and 425µL water) + 5µL cyanoborohydride 1M, heated at 55℃, 2 hours.  After the reaction, the solution was diluted in water to get a final volume of 50µL.  This solution was diluted in water 20x and 20,000x prior to CE-LEDIF analysis.

Method

PVA coated capillary:

65 cm x 50 µm ID

Buffer 40mM ε-aminocaproic acid pH 4.5 ajusted with pure acetic acid glacial + 0.02% hydroxypropylmethylcellulose

Voltage : 20kV

Injection : 0.5psi, 10s

Temperature : 20℃

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