Thin Layer Chromatography of aminoacids and short peptides


Below is a group TLC of all 20 aminoacids plus some other related compounds. TLC was done on a regular silica plate. 1uL of aminoacid solution in water (5mg/mL; 3mg Na2CO3 added for Cystine, Phenylalanine and Tyrosine) was applied (about 3mm spot was formed). This means each spot contained 5ug (65 nmols to 25 nmols) of an aminoacid. The plate was eluted with n-butanol:acetic acid:water 3:1:1(by volume) mixture followed by ninhydrin stain

TLC of aminoacids on silica stained with ninhydrin

Same aminoacids, same elutant but on cellulose TLC plate, ninhydrin stain.

TLC of aminoacids on cellulose stained with ninhydrin

Same aminoacids, same elutant but on neutral alumina TLC plate, ninhydrin stain.

TLC of aminoacids on alumina stained with ninhydrin

Structures are given for natural L(s-) aminoacids

LaneAminoacidRf
silica
(cell.)
FW:letter
codes
structure LaneAminoacidRf
silica
(cell.)
FW:letter
codes
structure
1Alanine0.30 (0.46)89.09ala

A
alanine structure 14Methionine0.51 (0.60)149.21met

M
Methionine structure
2Asparagine0.21 (0.24)132.12asn

N
asparagine structure 15Norleucine0.63 (0.69)131.17nleNorleucine structure
3Aspartic Acid0.24 (0.29)133.10asp

D
asparagine structure 16Phenylalanine0.62 (0.66)165.19phe

F
Phenylalanine structure
4Arginine0.16 (0.32)174.20arg

R
Arginine structure 17Proline0.24 (0.48)115.13pro

P
Proline structure
5Cysteine0.37 (0.41)121.16cys

C
Cysteine structure 18Hydroxyproline0.24 (0.31)131.13hypHydroxyproline structure
6Cystine0.14 (0.15)240.30-

C
Cystine structure 19Serine0.26 (0.25)105.09ser

S
Serine structure
7Glutamine0.25 (0.29)146.14gln

Q
Cystine structure 20Threonine0.30 (0.36)119.12thr

T
Threonine structure
8Glutamic Acid0.31 (0.35)147.13glu

E
Glutamic Acid structure 21Tryptophan0.61 (0.57)204.23trp

W
Tryptophan structure
9Histidine0.12 (0.32)155.15his

H
Histidine structure 22Tyrosine0.55 (0.44)181.19tyr

Y
Tyrosine structure
10Isoleucine0.53 (0.73)131.17lle

I
Isoleucine structure 23Valine0.44 (0.66)117.15val

V
Valine structure
11Glycine0.25 (0.26)75.07gly

G
Glycine structure 242,3-diamino propinic acid
(DAP)
0.10 (0.17)104.11dapDAP structure
12Lysine0.12 (0.24)146.19lys

K
Lysine structure 25Gly-Gly-Gly
tripeptide
0.18 (0.34)189.17-gly-gly-gly structure
13Leucine0.61 (0.71)131.18leu

L
Leucine structure 26Ser-Gly
dipeptide
0.18 (0.40)162.15-ser-gly structure
------ 27Ala-ala
dipeptide
0.23 (0.70)160.17-ala-ala structure