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Sugar Stock solutions
Sugar
MW (mg/mMol) Mass (mg/100mL) Purity (%)
D(+) galactose
180.17
100
99
Lactosemonohydrate 342
100
95
Internal Standard MW (mg/mMol) Mass (mg/100mL) Purity (%)
D(-) Ribose
180.17
30
99
Prepare the stock solutions according to the table above byweighing 100± 5mg in
a 100mL volumetric flask. Dissolve the components inMilli-Qwater and adjust to
100mL withMilli-Qwater. The concentration of the stock solution is 1mg/mL. Store
1mL portions in 2mL safelock tubes at -20°C
Standard series:
Make from the standard sugar stocks the following dilutions in 50mL volumetric
flasks:
Concentration
Galactose
(µg/mL)
Concentration
Lactose
(µg/mL)
Volume stock
Galactose (mL)
Volume stock
Lactose (mL)
Volume
Internal
Standard (mL)
30
25
1.50
1.25
5.00
24
20
1.20
1.00
5.00
12
10
0.60
0.50
5.00
6
5
0.30
0.25
5.00
0
0
0
0
5.00
Add galactose, lactose and ribose stock to the volumetric flasks and adjust to 50mL.
40mMPotassiumPhosphate pH6.0±0.1with30µg/mLRibose:
Pipet 5.00mL of 0.2M Potassium Phosphate pH 6.0 in a volumetric flask of 25mL.
Pipet 2.50mL of the Ribose stock. Adjust to 25mL with Milli-Q andmixwell.
Carrez treated lactose standard60µg/ml:
Pipet 3ml lactose stock (1mg/ml) in a 50mL volumetric flask. AddMilli-Qwater
until ¾ of the flask is filled. Pipet 750 µl of carrez 1 and 750 µl of carrez 2 to the
flask andmixwell. Adjust the volume until 50ml withMilli-Qwater, homogenize and
centrifuge for 10minutes at 13000 rpm. Fill a HPLC vial with the supernatant. This
standard is used to determine the fractionation time.
GOS-02
FORWORKINGGROUP/ERPUSEONLY
DONOTDISTRIBUTE
