[PDF] [PDF] Technical Support Information Bulletin 1073 - Aapptec

DMF in Acetic Anhydride - Home - Thieme Connect

DMF in Acetic Anhydride: A Useful Reagent for Multiple-Component Syntheses of sation system of DMF in acetic anhydride and highly acidic 1,2,5,6-

[PDF] Supporting Information - The Royal Society of Chemistry

published protocol 1 Dimethylformamide (DMF) stored over molecular sieves ( Labscan, Acetic anhydride, t-butanol, trifluoroacetic acid, sodium hydroxide and

[PDF] Standard practices for Fmoc-based solid-phase - UCI Chemistry

19 sept 2018 · It is incredibly important the you use the ”good” DMF Prepare the capping solution by combining acetic anhydride and pyridine in a 3:2 ratio 

A comparative study of terminating agents for use in solid-phase

acetic anhydride and triethylamine in DMF - in the solid-phase synthesis of H-Leu -Ala-Gly-Val-OH The terminating agent was used to block any N-terminal 

[PDF] Technical Support Information Bulletin 1073 - Aapptec

If DMF is used as the solvent in the coupling reaction, it should be degassed end-capped with acetic anhydride to block any unreacted active sites on the resin  

[PDF] Supporting Information - Wiley-VCH

NMM were added to a solution of 4 eq protected amino acid (0 1 M) in DMF DMF (5x) Capping: Acetic anhydride/pyridine (1:9) was added to the resin

[PDF] Synthesis and Purification of Peptide Nucleic Acids - Deep Blue

4 nov 2018 · Deblocking solution: 20 (v/v) piperidine in N,N-dimethylformamide (DMF) Capping solution: 5 (v/v) acetic anhydride/6 (v/v) 2,6-lutidine in 

[PDF] acetic anhydride ir spectrum

[PDF] acetic anhydride is obtained by the reaction of acetic acid and

[PDF] acetic anhydride ka formula

[PDF] acetic anhydride lewis structure

[PDF] acetic anhydride line formula

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[PDF] acetic anhydride literature melting point

[PDF] acetic anhydride melting boiling point

[PDF] acetic anhydride molecular formula

[PDF] acetic anhydride molecular melting point

[PDF] acetic anhydride molecular weight

[PDF] acetic anhydride msds fisher

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[PDF] acetic anhydride msds pdf

aapptecTechnical Support

Information Bulletin 1073

Wang Resin

Wang resin is the standard peptide synthesis resin used with Fmoc-chemistry. The resin is acid labile and finished peptides can be easily cleaved by treatment with 50 % (v/v) TFA/DCM. These relatively mild cleavage conditions have made this resin popular also in solid phase organic synthesis. The first amino acid is attached to Wang resin using an activating agent such as dicyclohexylcarbodiimide (DCC) and a catalytic amount of 4-dimethylamino-pyridine (DMAP). These conditions can lead to partial epimerization of the amino acid, so HOBt is normally added to reduce racemization. Even with HOBt added, racemization can be a problem if the first amino acid is Cys or His. If DMF is used as the solvent in the coupling reaction, it should be degassed under vacuum or sparged with nitrogen first to remove any dimethylamine that may be contaminating it. The dimethylamine can remove the Fmoc group of the attached amino acid and

lead to C-terminal oligomeric impurities. After the first amino acid is attached, the resin should be

end-capped with acetic anhydride to block any unreacted active sites on the resin. Fmoc-amino acid fluorides can also be used to attach the first amino acid. This method is useful where racemization may be a problem.

Attachment of the First Amino Acid

A.DIC/HOBt Method

1.In a round bottom flask suspend the resin in 9:1 v/v CH2Cl2/DMF (approximately

15 mL per gram of resin).

2.In a separate flask dissolve 1.5 to 2.5 equivalents (relative to the resin) of the

Fmoc-amino acid in a minimum amount of DMF. Add the same equivalency of HOBt. Stir the mixture until the HOBt dissolves. If the HOBt doesn't dissolve completely, add just enough DMF to bring it into solution. Add this solution to the resin.

3.In a separate flask dissolve 0.1 equivalent (relative to the resin) of DMAP in a

minimum amount of DMF.

4.Add 1.0 equivalent (relative to the amino acid) of DIC to the resin mixture then

add the DMAP solution. Fit a drying tube onto the flask.

5. Agitate the mixture with a mechanical shaker for 2 to 3 hours at room

temperature. Add 2 equivalents (relative to the resin) of acetic anhydride and pyridine to the reaction flask and mix an additional 30 minutes at room temperature to end cap any unreacted hydroxyl groups on the resin.

6.Filter the resin in a fine porosity sintered glass funnel and wash it 3 times with

DMF, then 3 times with DCM, and finally 3 times with methanol. In each wash use enough solvent to slurry the resin. After the final methanol wash, dry the resin in vacuo to a constant weight. The substitution of the resin can be estimated from the weight gain of the resin. For an accurate measurement of the resin substitution, the amount of Fmoc released from a weighed sample of resin can be measured spectrophotometrically.

Cleavage of the Peptide From the Resin

A.TFA Proceedure1

1.Place the resin in a round bottom flask and add 20% piperidine in DMF until the

resin is just covered. Let the mixture stand for 30 minutes to remove the N- terminal Fmoc group.

2.Transfer the resin to a sintered glass funnel with fine porosity and apply vacuum.

Wash the resin 3 times with DMF. Slurry the resin in DCM three times to remove the DMF.

3. Slurry the resin in 50%TFA in DCM (v/v) containing scavengers as required by

the amino acid composition of the peptide. Swirl the mixture occasionally during the reaction time. The reaction time will depend on the amino acid composition of the peptide. If there are no Arg(Mtr) or Arg(Pmc) groups, cleavage will take 1.5 to

2 hours. If the peptide contains Arg(Pmc), allow 2 to 4 hours for cleavage. If the

peptide contains Arg(Mtr), over 6 hours may be required for complete removal of the Mtr group.

4.Filter the resin in a fine sintered glass funnel. Wash the resin 3 times with small

portions of TFA.

5.Combine the filtrates and add 8-10 times the volume of cold ether. If necessary,

keep the mixture at 4°C overnight to precipitate the peptide. Filter the peptide using a fine sintered glass funnel. Wash the crude peptide further with cold ether.

B.TMSBr Procedure2

If the peptide contains Arg(Mtr), this procedure will cleave the peptide from the resin and remove the Mtr group more rapidly than the TFA procedure.

1.Place the resin in a round bottom flask and add 20% piperidine in DMF until the

resin is just covered. Let the mixture stand for 30 minutes to remove the N- terminal Fmoc group.

2.Transfer the resin to a sintered glass funnel with fine porosity and apply vacuum.

Wash the resin 3 times with DMF. Slurry the resin in DCM three times to remove the DMF.

1 Based on results reported in Jubilut, GN; Cilli, EM; Crusca, E; Silva, EH; Okada, Y; Nakaie, CR.

Chem Pharm Bull (Tokyo) 2007, 55, 468-470.2 Based on procedures in Guo,S.; et al. Chem. Pharm. Bull. 1988, 36, 4989; Yajima, H.; et al.

Tetrahedron 1988, 44, 805-819.

3.For 100 mg of peptide-resin, mix 250 uL of ethanedithiol, 50 uL of m-cresol, 590

uL of thioanisole and 3.75 mL of TFA. Cool the mixture in an ice bath then add

660 ul of TMSBr. Cool the cleavage mixture to 0 ºC, then add 100 mg of the

peptide resin. Allow the mixture to stand 15 minutes under nitrogen.

4.Filter the resin in a fine sintered glass funnel. Wash the resin 3 times with small

portions of TFA.

5.Combine the filtrates and add 8-10 times the volume of cold ether. If necessary,

keep the mixture at 4 °C overnight to precipitate the peptide. Filter the peptide using a fine sintered glass funnel. Dissolve the precipitated peptide in 20% aqueous acetic acid and lyophilize.quotesdbs_dbs17.pdfusesText_23