LC separation was carried
out at room temperature on an Eurospher NH2 column, 100 Å pore
size, 7 mm particle size, 250 4.6 mm i.d. (Knauer, Berlin, Ger-many). Prior to use, solvents were filtered through a 0.45-mm
membrane filter and degassed for 15 min in an ultrasonic bath
(Cleaner Model SM 25E-MT, Branson Ultrasonics Corporation, Dan-bury, CT). The mobile phase was an 80:20 (v/v) mixture of acetoni-trile and ultrapure water. The LC was connected to a refractive
index detector K-2301 (Knauer). The flow rate and the injection
volume were 1 ml/min and 20 ll, respectively. Identified sugars
were quantified on the basis of peak areas of external standards
consisting of glucose (2%), fructose (2%) and sucrose (2%) solutions.
Total reducing sugars were obtained as the sum of glucose and
fructose values. Each sample was analysed in triplicate and quan-tification was carried out from integrated peak areas of the sample
against the corresponding standard graph. Results were expressed
as percentage of dry weight