Citric Acid → HCID 131724567, compound

HRID 1837280Notes

Reaction equation

HRID 1837280 reaction equation
S[C@@H:2]1[O:10][C@H:9]([CH2:11][OH:12])[C@H:7]([OH:8])[C@H:5]([OH:6])[C@H:3]1[OH:4].C(O)(=O)CC(CC(O)=O)(C(O)=O)[OH:16].[Na+].[Cl-].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O.[C:48]1(=[O:60])[N:52]([CH2:53][CH2:54][CH2:55][C:56]([NH2:58])=[O:57])[C:51](=[O:59])[CH:50]=[CH:49]1>C(O)C>[C:51]1(=[O:59])[N:52]([CH2:53][CH2:54][CH2:55][C:56]([NH2:58])=[O:57])[C:48](=[O:60])[CH:49]=[CH:50]1.[OH:16][CH:2]1[O:10][C@H:9]([CH2:11][OH:12])[C@H:7]([OH:8])[C@H:5]([OH:6])[C@H:3]1[OH:4]

Participants

7

Reactant 5

HCID 311Citric AcidHCID 5234Sodium ChlorideHCID 6049Ethylenediaminetetraacetic AcidHCID 17867096HCID 17867096, compound2×HCID 18638434HCID 18638434, compound

Product 1

HCID 131724567HCID 131724567, compound

Reagents, catalysts & solvents 1

HCID 702EthanolSolvent2×

Procedure

1-thio-b-galactopyranose (1.5 mg) is dissolved in 150 μl of buffer A (20 mM citric acid, 35 mM Na2HPO4, 108 mM NaCl, 1 mM ethylenediamine-tetraacetic acid (EDTA)) and 150 μl of ethanol, N-(m-(3-trifluoromethyl)diazirine-3-yl)phenyl)-4-maleimido-butyramide (MAD, 3.3 mg, preparation according to A. Collioud et al., Bioconjugate Chemistry 4, 528-536 (1993)) is dissolved in 150 μl of ethanol and an additional 150 μl of buffer A. Both solutions are mixed. The mixture is stirred and allowed to react at room temperature for 90 min, to give the compound MAD-Gal. The product is purified by HPLC. The HPLC flow rate is 2 ml/min. For gradient elution (0 min.-100% A; 5 min.-100% A; 20 min.-70% A; 30 min.-0% A; 40 min. 0%A), solvent B is added to solvent A (solvent A, 0.1% TFA in water; solvent B, 0.1% TFA in acetonitrile/water (4:1 by volume)). Detection is performed at 254 nm and fractions of the eluant are collected. 13C.NMR (D2O chemical shift (δ) in parts per million): 179.14, 178.40, 177.99, 177.86, 173.30, 137.79, 129.53, 129.38, 122.46, 121.84, 118.32, 84.42, 84.27, 78.99, 78.91, 73.84, 73.66, 69.73, 69.68, 68.58, 68.49, 60.80, 60.69, 39.61, 38.66, 38.46, 37.71, 35.83, 33.57, 22.68, 22.57 ppm. Mass Calc (M−H): 561.1 Da/e, found: 561.1 Da/e.

Workup

  1. additionBoth solutions are mixed
  2. customto react at room temperature for 90 min

Sources & evidence

ORD record
ord-bdc84f4dbb1d421d99a176ec4e8b2e29
Source dataset
uspto-grants-2004_08
Patent number
US06780980B1

Reaction notes

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