Ethyl-3- 3-dimethylaminopropyl carbodiimide Hydrochloride

InChl = l/C8H17N3.ClH/cl-4-9-8-10-6-5-7-ll(2)3 /h4-7H2, l-3H3 lH/fC8H18N3.Cl/hllH lh/q+l -l InChIKey = FPQQSJJWHUJYPU-CZVSVQNTCV (base) 

Alternate Name l-(3-dimethylaminopropyl)-3-ethylcarbodi-imide water-soluble carbodiimide EDC EDCI. 

Physical Data free base is an oil, bp 47-48 °C/0.27 nrniHg HCl salt is a white powder, mp 111-113 °C Mel salt mp 97-99 °C. 

Form Supplied in commercially available as HCl salt and as me-thiodide salt that are white solids. Reagents are 98% pure main impurity is the urea that can form upon exposure to moisture. 

Analysis of Reagent Purity IR 2150 cm (N=C=N stretch) urea has C=0 stretch near 1600-1700 cm.  

Freidel, C. Crafts, J. A. LA 1884, 1, 527 see also The Chemistry of Functional Groups—The Chemistry of Acyl Halides, S. Patai, Ed. Interscience New York, 1972. 

Handling, Storage, and Precautions EDC is moisture-sensitive store under N2 in a cool dry place. It is incompatible with strong oxidizers and strong acids. EDC is a skin irritant and a contact allergen therefore avoid exposure to skin and eyes. 

Ftepttde Coupling Reagent. This carbodiimide (EDC) reacts very similarly to 1,3-Dicyclohexylcarbodiimide and other car-bodiimides. The advantage EDC has over DCC is that the urea produced is water soluble and, therefore, is easily extracted. Di-cyclohexylurea is only sparingly soluble in many solvents and is removed by filtration which may not be as effective as extraction. A typical example of EDC for peptide coupling is shown in (eq 1)  

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COOH groups of the PE (PAA or PGA) and the -NH2 moieties of the pre-loaded protein, using l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) as a catalyst [99]. Negligible enzyme desorption (<0.1 %) is observed from cross-linked lysozyme-loaded MS spheres after exposing the samples to an aqueous solution for 48 h, while about 25 % of the immobilized lysozyme is desorbed under the same conditions when the lysozyme is not cross-linked. 

The reactions involved in an EDC-mediated conjugation are discussed in Chapter 3, Section 1.1 (Note EDC is l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride MW 191.7 and is sometimes referred to as EDAC). The carbodiimide first reacts with available carboxylic groups on either the carrier or hapten to form a highly reactive o-acylisourea intermediate. The activated carboxylic group then can react with a primary amine to form an amide bond, with release of the EDC mediator as a soluble isourea derivative. The reaction is quite efficient with no more than 2 hours required for it to go to completion and form a conjugated immunogen. 

Weigh out 1.25mg of the carbodiimide EDC (l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride Thermo Fisher) into a microfuge tube. 

AT-Acyl-L-HSL can be conveniently prepared in excellent yields by the acylation of l-HSL either with the corresponding carboxylic acids activated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) in a water/ 1,4-dioxane (1 1) solvent system or with the corresponding acid chlorides in dichloromethane (DCM) in the presence of triethylamine (Scheme 4) [15,16, 37,53]. 

NPH HCl) with l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC HCl) as a coupling agent [593,594], The advantage is that with hydrazines it is possible to carry out the derivatization in milder conditions. An efficient postcolumn derivatization is obtained using avidin or streptavidin. In fact, these two proteins react in a very specific way with biotin, therefore they are bound with a fluorescent marker, such as fluorescein 5-isothiocyanate (FITC) to obtain fluorescent derivatives. Some authors [595] report the use of MS or MS/MS for biotin detection, but this method seems to be less sensitive than FLD. 

The carbodiimide of choice used to couple cystamine to carboxylate- or phosphate-containing molecules is most often the water-soluble carbodiimide EDC (l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride Chapter 3, Section 1.1). This reagent rapidly reacts with carboxylates or phosphates to form an active complex highly reactive toward primary amines. The reaction is efficient from pH 4.7 to 7.5, and a variety of buffers may be used, providing they do not contain competing groups. 

Add EDC (l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride Pierce) to a final concentration of 2 mg/ml in the reaction solution. To aid in the addition of a small amount of EDC, a higher concentration stock solution may be prepared in water and an aliquot added to the reaction to give the proper concentration. Since EDC is labile in aqueous solutions, the stock solution must be made quickly and used immediately. 

A mixture of 4 -[(methylsulfonyl)amino]-y-oxobenzenebutanoic acid and 1-hydroxybenzotriazole in dimethylformamide (DMF) under nitrogen, is treated with dibutylamine in DMF. The mixture is cooled in an ice bath and l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) added in portions over 5 min. The mixture is stirred in the cold 1 h and overnight at room temperature. The solvent is removed in vacuo (bath temperature 35°C). The residue is treated with ice and ethyl acetate and the organic layer washed sequentially with 0.5 N monopotassium sulfate, cold 4% NaHC03, cold water and finally brine. The organic solution is dried (Na2S04) and concentrated in vacuo. The N,N-dibutyl-y-oxo-4-[(methylsulfonyl)amino]benzenebutan-amide (crystallized from ethyl acetate-hexane) is obtained. 

Abbreviations used Dde, N-[l-(4,4-dimethyl-2,6-dioxocyclohexadiene)]-ethyl DIEA, diisopropylethylamine DMF, N,N-dimethylformamide EDC, l-Ethyl-3-(3 -dimethylaminopropyl)carbodiimide hydrochloride HOBt, 1-hydroxybenzotriazole NHS, N-hydroxysuccinimide NMP, l-methyl-2-pyrroIidinone PAL, Peptide Amide Linker [5-(4-(9-fluorenylmethyloxycarbonyl) aminomethyl-3,5-dimethoxyphenoxy) valeric acid] PyBOP, Benzotriazole-l-yl-oxy-trispyrrolidino-phosphonium hexafluorophosphate TFA, trifluoroacetic acid. 

The Ox group can be removed by reduction, either by palladium-catalyzed hydrogenation (quantitative) or by use of sodium in liquid ammonia (75-85 % yield). The Ox group can also be cleaved by oxidation with excess m-chloroperbenzoic acid followed by hydrolytic workup (70%). Simple Ox dipeptides have been prepared using 1-ethyl-3-(3 -dimethylaminopropyl)carbodiimide hydrochloride (1, 371). The free dipeptides... 

Modification of the protein amine groups is the procedure most frequently used to produce antibody-liposome conjugates. Early procedures used crosslinking agents, such as l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) (27,28) in the presence of preformed liposomes containing a lipophilic carboxylic acid. Condensing agents like these tend to form protein-protein polymers. Control of these reactions is typically difficult and complex, and as a result separation of the liposomes from protein polymers is a... 

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