Glucose derivative: Aldonitrile acetat
Preparation of glucose aldonitrile acetate.
1. Hydroxyl amine hydrochloride -- Sigma H-2391 (ACS reagent.) Keep in desiccator.
2. Acetic anhydride 每
Supelco 3-3085
3. Pyridine 99.9% HPLC grade 每 Sigma-Aldrich 27,040-7
4. Ethyl acetate 每 Fisher OPTIMA E196-4
Standards: unlabeled glucose, pure labeled standard.
Procedure:
1. Prepare glucose 每 deproteinized, deionized, lyophilized, 50-200 mg glucose.
2. Prepare hydroxylamine
hydrochloride solution in pyridine, 20 mg / 1 ml.
3. Add 100 mL of hydroxylamine solution to each tube. Heat to 100oC for 30 min in heating block.
4. Add 75 mL of acetic anhydride, heat for 1 hour, 100 oC.
5. Evaporate to dryness with N2.
6. Add 50-200mL of ethyl acetate as solvent just before GC/MS analysis.
GC/MS:
GC column: HP5, length 30m, 250mm ID.
Inlet temp. 250oC, split ratio 1:100 adjusted depending on sample concentration.
Relevant ion clusters
for SIM analysis:
CI mode, m/z 328 cluster for C1-C6. (monitor m/z 327 to m/z 336)
EI mode: m/z 187 cluster for C3-C6 fragment,
and m/z 242 cluster for C1-C4 fragment.
Reagents:
1. Acetic anhydride 每 Supelco 3-3085
2. Pyridine 99.9% HPLC grade 每 Sigma-Aldrich 27,040-7
3. Ethyl acetate 每 Fisher OPTIMA E196-4
Procedure:
1. To a dry glucose sample add 75 mL of pyridine and 75 mL acetic
anhydride, heat for 1 hour, 100oC.
2. Evaporate to dryness with N2.
1. Add 50-200mL of ethyl
acetate as solvent just before GC/MS analysis.
GC/MS:
GC column: HP5, length 30m, 250mm, programmed at constant He flow
of 1ml/min.
Inlet temp. 250oC, split ratio 1:100 adjusted depending on sample concentration.
Relevant ion clusters for SIM analysis:
CI mode, m/z 169 cluster and m/z 331 cluster for C1-C6.
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