Amide Coupling Array

Carbodiimide Coupling Reagent Kit (HCK1005-01-002)

2 sets of 4 reagents; DCC, DIC, EDC (HCl) and ethyl(hydroxyimino)cyanoacetate 8 reaction vials total
This kit requires EvoluChem reaction block (HCK1006-01-002) included in the EvoluChem Starter Kit (HCK1006-01-001)

The typical protocol is performed at 0.1 mol/l concentration reaction condition using a solution of combined acid and amine. However you can a different ratio of amine and acid. Recommended solvents are CH2Cl2, DMF or NMP. Solvent should be selected based on solubility of the substrates. Recommended bases are Et3N or DIPEA.

Protocol at 100 µl volume reaction condition

  1. Prepare required volume of a solution of acid substrate 0.2 mol/L in the desired solvent. For example, 250 µl solution for 4 reaction conditions (50 µl extra to compensate potential evaporation).
  2. Using a clean and dry syringe, add 50 µl of the acid solution to each reaction vial containing a coupling reagent.
  3. Add 10 µl of the solution of ethyl (hydroxyimino)cyanoacetate to each reaction vial. (prepared by 70 µl of DMF in the reagent vial)
  4. Stir the reaction vials in the reaction block at desired temperature (between 0°C and 25°C) for 30 min.
  5. Prepare 250 µl of a solution of amine substrate 0.2 mol/L and base (Et3N or DIPEA) 0.4 mol/L in the desired solvent. Add 50 µl of this solution to each reaction vial.
  6. Stir the reaction vials in the reaction block at desired temperature for 3 to 12 hours. Do not heat vials at temperature higher than 120°C or higher than 5°C above the boiling point of the used solvent.
  7. Remove the vial caps using a decapper.
  8. Prepare analytical sample for each reaction condition with 5 µl sample diluted into 200 µl in either DMSO or water/acetonitrile 50/50. Analyze resulting analytical samples by LC/MS.

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