pellets dissolve. Add increments of water to help the pellets
dissolve. Bring to volume with water. Mix well in stoppered
flask. Store solution at room temperature in a plastic (HDPE)
container, and remake once a precipitate forms (looks like
silvery flakes)—typically this happens after 2–3 weeks. Sodium
hydroxide is a moderate health hazard.
4. Prepare a 3 mg/mL MBTH (3-methyl-2-benzothiazolinone
hydrazine) solution in water, and store in the dark at 4
C for
up to 1 week. MBTH is a moderate health hazard.
5. Prepare a 1 mg/mL DTT (dithiothreitol) solution in water,
and store in the dark at 4
C for up to 1 week. Dithiothreitol is
a moderate health hazard with mild flammability and reactivity.
6. Prepare 0.25 M HCl (hydrochloric acid) in a 200 mL volumetric flask, and store at room temperature in the dark for up to
1 month. Hydrochloric acid is a moderate health hazard. Add a
small amount of water to the volumetric flask. Pipet 4.16 mL of
concentrated HCl into the flask. Bring to volume with water
and cap, and mix thoroughly.
7. Ferric solution: 0.5% ferric ammonium sulfate dodecahydrate,
0.5% sulfamic acid (w/v), and 0.25 M HCl. Dissolve 200 mg of
ferric ammonium sulfate dodecahydrate and 200 mg of sulfamic acid into 40 mL of 0.25 M HCl. This solution may be
stored at room temperature in the dark for up to 1 month.
Sulfamic acid and ammonium ferric sulfate dodecahydrate are
moderate health hazards.
2.5 Liquid
Chromatography
Reagents and
Standards
Use 18.2 MΩ water for all preparations requiring water. Carefully
follow all relevant chemical handling and disposal procedures.
1. 72% (w/w, specific gravity 1.6338 at 20
C) sulfuric acid, used
for sample two-step hydrolysis.
2. Prepare a master mix of all the sugars [L(À)fucose, L(+)rhamnose monohydrate, L(+)arabinose, D(+)galactose, D(+)glucose,
D(+)mannose, D(+)xylose, D(À) ribose, D(+)galacturonic acid
monohydrate, D(À)glucuronic acid] in a 10 mL, class A, volumetric flask by weighing out each sugar at ~ 20 mg (all sugars
should be weighed into the same volumetric flask). Record the
exact weight of each sugar to 0.1 mg. Add approximately 5 mL
water to the volumetric flask, and swirl or vortex to dissolve all
sugars. Bring to volume with water, and mix well in the stoppered flask. Transfer 1 mL aliquots of the ~2 mg/mL master
sugar mix into vials that may be sealed and frozen. The master
sugar mix vials may be stored for up to 2 months in a freezer
(À20
C).
3. Follow Thermo Fisher Scientific/Dionex TN 71 instructions
to prepare 100 mM NaOH and 1 M sodium acetate (NaOAc)/
100 mM NaOH eluent solutions. Store in plastic (high-density
polyethylene, HDPE) solvent bottles.
194
S. Van Wychen and L.M.L. Laurens
dissolve. Bring to volume with water. Mix well in stoppered
flask. Store solution at room temperature in a plastic (HDPE)
container, and remake once a precipitate forms (looks like
silvery flakes)—typically this happens after 2–3 weeks. Sodium
hydroxide is a moderate health hazard.
4. Prepare a 3 mg/mL MBTH (3-methyl-2-benzothiazolinone
hydrazine) solution in water, and store in the dark at 4
C for
up to 1 week. MBTH is a moderate health hazard.
5. Prepare a 1 mg/mL DTT (dithiothreitol) solution in water,
and store in the dark at 4
C for up to 1 week. Dithiothreitol is
a moderate health hazard with mild flammability and reactivity.
6. Prepare 0.25 M HCl (hydrochloric acid) in a 200 mL volumetric flask, and store at room temperature in the dark for up to
1 month. Hydrochloric acid is a moderate health hazard. Add a
small amount of water to the volumetric flask. Pipet 4.16 mL of
concentrated HCl into the flask. Bring to volume with water
and cap, and mix thoroughly.
7. Ferric solution: 0.5% ferric ammonium sulfate dodecahydrate,
0.5% sulfamic acid (w/v), and 0.25 M HCl. Dissolve 200 mg of
ferric ammonium sulfate dodecahydrate and 200 mg of sulfamic acid into 40 mL of 0.25 M HCl. This solution may be
stored at room temperature in the dark for up to 1 month.
Sulfamic acid and ammonium ferric sulfate dodecahydrate are
moderate health hazards.
2.5 Liquid
Chromatography
Reagents and
Standards
Use 18.2 MΩ water for all preparations requiring water. Carefully
follow all relevant chemical handling and disposal procedures.
1. 72% (w/w, specific gravity 1.6338 at 20
C) sulfuric acid, used
for sample two-step hydrolysis.
2. Prepare a master mix of all the sugars [L(À)fucose, L(+)rhamnose monohydrate, L(+)arabinose, D(+)galactose, D(+)glucose,
D(+)mannose, D(+)xylose, D(À) ribose, D(+)galacturonic acid
monohydrate, D(À)glucuronic acid] in a 10 mL, class A, volumetric flask by weighing out each sugar at ~ 20 mg (all sugars
should be weighed into the same volumetric flask). Record the
exact weight of each sugar to 0.1 mg. Add approximately 5 mL
water to the volumetric flask, and swirl or vortex to dissolve all
sugars. Bring to volume with water, and mix well in the stoppered flask. Transfer 1 mL aliquots of the ~2 mg/mL master
sugar mix into vials that may be sealed and frozen. The master
sugar mix vials may be stored for up to 2 months in a freezer
(À20
C).
3. Follow Thermo Fisher Scientific/Dionex TN 71 instructions
to prepare 100 mM NaOH and 1 M sodium acetate (NaOAc)/
100 mM NaOH eluent solutions. Store in plastic (high-density
polyethylene, HDPE) solvent bottles.
194
S. Van Wychen and L.M.L. Laurens