J.
Calculations
Calculate themolar concentration for eachOSpeakusing themaltotriose calibration curveusing
equation1:
Equation1
m
Amt_IS
A
A_IS
Amt_IS
C
A_IS
A_OS
C_OS
sple
sple
std
std
std
std
sple
sple
sple
V
.
.
.
.
Where:
C_OS
sple
=Oligosaccharidemolar concentration in the sample test portion inµmol/g.
C
std
= Concentrationofmaltotriose in standard solution (6.2) inµmol/mL.
Amt_IS
sple
=Amount of Laminaritriose ISadded to the test sample inµmol.
Amt_IS
std
=Amount of Laminaritriose ISadded to the calibration standard inµmol.
A_OS
sple
=Areaof theOSpeak in the sample injection.
A_
std
=Areaofmaltotriosepeak in the standard injection.
A_IS
sple
= Areaof the ISpeak in the sample injection.
A_IS
std
=Areaof the ISpeak in the standard injection.
V=Volumeof sampleextract inmL.
m
sple
=Massof the sample test portion inmg.
Note:
For thepurposeof thismethod,molar response factorsof thedifferent oligosaccharides are
considered tobeequivalent tomaltotriose. Calibrate the fluorescencedetectorwithmaltotriose
standard anduse thismolar response toobtainamolar concentrationof eacholigosaccharide.
ExpressionofOS concentration ing/100 g isobtainedbymultiplying themolar concentration
(obtainedwithequation1) by themolecularweight of theoligosaccharideof interest. This canbe
doneby runningaprofileof theGOSusing the samemethodologywitha LC-FLD-MS instrument. The
totalGOS is then cacluatedby summing these together as follows:-
w = Σ (OSmolar conc. x MW ) x0.0001
Where:
w=amount ofGOS in the sample ing/100g
GOS-01
FORWORKINGGROUP/ERPUSEONLY
DONOTDISTRIBUTE
