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== TNBS Test - Primary Amines == | |||
== TNBS Test - | '''Solution 1''': 10% DI(P)EA in DMF (store at 25<sup>o</sup> C)<br> | ||
'''Solution 1''': 10% DI(P)EA in DMF (store at 25<sup>o</sup>)<br> | '''Solution 2''': 1 M aqueous TNBS (store at 4<sup>o</sup> C)<br><br> | ||
'''Solution 2''': 1 M aqueous TNBS (store at 4<sup>o</sup>)<br><br> | |||
'''Procedure''':<br> | '''Procedure''':<br> | ||
- Place a few resin beads in a small test tube.<br> | - Place a few resin beads in a small test tube.<br> | ||
Line 16: | Line 15: | ||
|- | |- | ||
|Intense orange/red beads||Yellow/light orange beads/bead core||Colourless beads | |Intense orange/red beads||Yellow/light orange beads/bead core||Colourless beads | ||
|} | |||
== Kaiser Test - Primary Amines == | |||
'''WARNING''': Ambiguous with Wang resins<br> | |||
'''Solution 1''': 5 g ninhydrin in 100 mL ethanol (store at 25<sup>o</sup> C, in dark)<br> | |||
'''Solution 2''': 80 g phenol in 20 mL ethanol (store at 25<sup>o</sup> C, in dark)<br> | |||
'''Solution 3''': 2 mL 0.001 M KCN in 98 mL pyridine (store at 25<sup>o</sup> C, in dark)<br><br> | |||
'''Procedure''':<br> | |||
- Place a few resin beads in a small test tube.<br> | |||
- Wash resin with ethanol 2-3 times to remove adsorbed DMF<br> | |||
- Add 2-5 drops of each solution<br> | |||
- Mix, heat to 100<sup>o</sup> C for 5 minutes<br> | |||
{| border="2" | style="font-size:120%; text-align:center; margin: 1em auto 1em auto;" | |||
|- | |||
! Free Amine (positive) | |||
! Ambiguous/Partial Positive | |||
! Negative | |||
|- | |||
|Variable intensity blue solution/resin||Blue bead cores||Colourless/light yellow beads | |||
|} | |||
== Chloranil Test - Primary Amines (acetaldehyde) - Secondary amines (acetone)== | |||
'''WARNING''': Keep solution for a maximum of one month<br> | |||
'''Solution 1''': 2% acetaldehyde (primary amines) or 2% acetone (secondary amines) in DMF (store at 4<sup>o</sup> C)<br> | |||
'''Solution 2''': 2% chloranil in DMF (store at 4<sup>o</sup> C)<br><br> | |||
''ALTERNATIVE SOLUTIONS''<br> | |||
'''Solution 1''': Saturated solution of chloranil in toluene (store at 4<sup>o</sup> C)<br> | |||
'''Solution 2''': Only necessary for primary amines. 2% acetaldehyde DMF (store at 4<sup>o</sup> C)<br><br> | |||
'''Procedure''':<br> | |||
- Place a few resin beads in a small test tube.<br> | |||
- Add 2-5 drops of each solution<br> | |||
- Mix, leave at room temperature for 5 minutes<br> | |||
{| border="2" | style="font-size:120%; text-align:center; margin: 1em auto 1em auto;" | |||
|- | |||
! Free Amine (positive) | |||
! Ambiguous/Partial Positive | |||
! Negative | |||
|- | |||
|Dark blue/green beads||Blue/green bead cores||Colourless/light yellow beads | |||
|} | |||
== Bromophenol Blue Test - Primary and Secondary amines == | |||
'''WARNING''': Ensure resin is well washed with DMF before performing this test<br> | |||
'''Solution 1''': 0.05% bromophenol blue in DMA (store at 25<sup>o</sup> C)<br><br> | |||
'''Procedure''':<br> | |||
- Place a few resin beads in a small test tube.<br> | |||
- Add 10-20 drops of solution<br> | |||
- Inspect immediately<br> | |||
{| border="2" | style="font-size:120%; text-align:center; margin: 1em auto 1em auto;" | |||
|- | |||
! Free Amine (positive) | |||
! Ambiguous/Partial Positive | |||
! Negative | |||
|- | |||
|Blue beads||---||Colourless beads | |||
|} | |||
== Isatin Test - Secondary amines == | |||
'''Solution 1''': Saturated isatin in benzyl alcohol with BOC-Phe-OH<br> | |||
'''Preparation''':<br> | |||
- Saturated isatin (~0.4 g) in benzyl alcohol (~12 mL)<br> | |||
- Stir for ~2 hours at room temperature, filter remaining isatin<br> | |||
- Mix 10 mL of filtered solution with 0.5 g BOC-Phe-OH<br><br> | |||
'''Procedure''':<br> | |||
- Place a few resin beads in a small test tube.<br> | |||
- Add 3-4 drops of solution<br> | |||
- Heat to 100<sup>o</sup> C for a 1.5 to 2 minutes ('''MAXIMUM''')<br> | |||
{| border="2" | style="font-size:120%; text-align:center; margin: 1em auto 1em auto;" | |||
|- | |||
! Free Amine (positive) | |||
! Ambiguous/Partial Positive | |||
! Negative | |||
|- | |||
|Blue beads||---||Colourless beads | |||
|} | |} |
Latest revision as of 07:51, 7 July 2010
TNBS Test - Primary Amines
Solution 1: 10% DI(P)EA in DMF (store at 25o C)
Solution 2: 1 M aqueous TNBS (store at 4o C)
Procedure:
- Place a few resin beads in a small test tube.
- Add 1-3 drops of each solution
- Mix, and leave at room temperature for 10 minutes
Free Amine (positive) | Ambiguous/Partial Positive | Negative |
---|---|---|
Intense orange/red beads | Yellow/light orange beads/bead core | Colourless beads |
Kaiser Test - Primary Amines
WARNING: Ambiguous with Wang resins
Solution 1: 5 g ninhydrin in 100 mL ethanol (store at 25o C, in dark)
Solution 2: 80 g phenol in 20 mL ethanol (store at 25o C, in dark)
Solution 3: 2 mL 0.001 M KCN in 98 mL pyridine (store at 25o C, in dark)
Procedure:
- Place a few resin beads in a small test tube.
- Wash resin with ethanol 2-3 times to remove adsorbed DMF
- Add 2-5 drops of each solution
- Mix, heat to 100o C for 5 minutes
Free Amine (positive) | Ambiguous/Partial Positive | Negative |
---|---|---|
Variable intensity blue solution/resin | Blue bead cores | Colourless/light yellow beads |
Chloranil Test - Primary Amines (acetaldehyde) - Secondary amines (acetone)
WARNING: Keep solution for a maximum of one month
Solution 1: 2% acetaldehyde (primary amines) or 2% acetone (secondary amines) in DMF (store at 4o C)
Solution 2: 2% chloranil in DMF (store at 4o C)
ALTERNATIVE SOLUTIONS
Solution 1: Saturated solution of chloranil in toluene (store at 4o C)
Solution 2: Only necessary for primary amines. 2% acetaldehyde DMF (store at 4o C)
Procedure:
- Place a few resin beads in a small test tube.
- Add 2-5 drops of each solution
- Mix, leave at room temperature for 5 minutes
Free Amine (positive) | Ambiguous/Partial Positive | Negative |
---|---|---|
Dark blue/green beads | Blue/green bead cores | Colourless/light yellow beads |
Bromophenol Blue Test - Primary and Secondary amines
WARNING: Ensure resin is well washed with DMF before performing this test
Solution 1: 0.05% bromophenol blue in DMA (store at 25o C)
Procedure:
- Place a few resin beads in a small test tube.
- Add 10-20 drops of solution
- Inspect immediately
Free Amine (positive) | Ambiguous/Partial Positive | Negative |
---|---|---|
Blue beads | --- | Colourless beads |
Isatin Test - Secondary amines
Solution 1: Saturated isatin in benzyl alcohol with BOC-Phe-OH
Preparation:
- Saturated isatin (~0.4 g) in benzyl alcohol (~12 mL)
- Stir for ~2 hours at room temperature, filter remaining isatin
- Mix 10 mL of filtered solution with 0.5 g BOC-Phe-OH
Procedure:
- Place a few resin beads in a small test tube.
- Add 3-4 drops of solution
- Heat to 100o C for a 1.5 to 2 minutes (MAXIMUM)
Free Amine (positive) | Ambiguous/Partial Positive | Negative |
---|---|---|
Blue beads | --- | Colourless beads |