Download p-Anisaldehyde: This clear tan stain is prepared by dissolving 0

TLC 용 발색 시약 제조법 및 용도
1. p-Anisaldehyde: This clear tan stain is prepared by dissolving 0.7 mL of p-anisaldehyde in 250 mL of
EtOH containing 9.5 mL of conc. H2SO4 and 2.7 mL of acetic acid. The plate is developed by heating on a
hot plate. This stain chars lavender and is good for oxygenated compounds.
2. Cobalt(II) chloride: This pink stain is prepared by dissolving ~2 g of CoCl2 in 100 mL of water and 10
mL of conc. H2SO4. The stain is used in conjunction with PMA. In cases where PMA is not effective
enough, dipping the plate (already processed with PMA) in CoCl 2 assists the staining process. The spots
are often different colors against a pink or colorless background.
3. 2,4-Dinitrophenylhydrazine: This stain is prepared by dissolving 12 g of 2,4-dinitrophenylhydrazine in
200 mL of EtOH, 60 mL of conc. H2SO4 and 80 mL of water. The stain is effective for carbonyl containing
compounds.
4. Ehrlich's Reagent: This stain is prepared by dissolving 1.0 g of p-dimethylbenzaldehyde in 75 mL of
MeOH and 50 mL of concentrated HCl. The stain is effective for indoles, amines, ergot alcohols and
pyrrolizidine alkaloids.
5. Ceric Ammonium Molybdate (CAM): This stain is prepared by dissolving 2.5 g of (NH4) 6-Mo7O24
and 1.0 g Ce(SO4)2 in 90 mL of water and 10 mL of conc. H2SO4. The plate is developed by heating on a
hot plate. This stain chars blue and is good for polyhydroxylated compounds.
6. I2: A jar of I2 crystals or a jar of fine grade silica gel impregnated with iodine works well. The plate is
simply allowed to stand in the jar of iodine vapor or is immersed in the silica gel for a few seconds. After
examining the plate, the stain can be removed by heating the plate briefly on a hot plate. The plate can then
be stained by other means.
7. Ninhydrin: This stain is prepared by dissolving 0.3 g of ninhydrin in 100 mL of n-BuOH or EtOH
containing 1-3 mL of acetic acid. The plate is developed by heating on a hot plate and spots are shades of
purple or brown. The stain is specific for amino acids, amino sugars and primary, secondary, and tertiary
amines.
8. Phosphomolybdic acid (PMA): This green-yellow stain is prepared by dissolving ~3-4 g of PMA in
100 mL of EtOH. The plate is developed by heating on a hot plate. PMA is the most commonly used
stain for organic materials, since it reacts with a wide variety of functional moieties. The spots are usually
black against a yellow-green background. PMA is also effective with extremely dilute reaction samples and
in this regard can be used semi-quantitatively.
9. Potassium Permanganate: This stain is prepared by dissolving 3 g of KMnO4 and 20 g of K2CO3 in 5
mL of 5% NaOH and 300 mL of water.
10. Vanillin: This light tan stain is prepared by dissolving 1-2 g of vanillin in 100 mL of EtOH containing
1 mL of conc. H2SO4. The plate is developed by heating on a hot plate. This stain is particularly good for
hydroxyl and carbonyl containing molecules. The spots are often brightly colored. This may be an
advantage when two molecules have the same Rf value.
TLC Stains
p-Anisaldehyde (1L 병 기준)
Ninhydrin
EtOH 95% 800 mL
Dissolve 0.3 g ninhydrin in 100 mL
p-Anisaldehyde 40 mL
of n-butanol ; add 3 mL AcOH
H2SO4 40 mL
Acetic acid 0.8 mL
Phosphomolybdic Acid (PMA)
� 0℃ Ice bath 에서 위 순서대로
10% (w/v) PMA solution in ethanol
가능한 서서히 시약 가한 뒤, 냉장보관.
CuSO4
3% (w/v) CuSO4 in 15% aq. H3PO4
KMnO4
DNP
1.5 g KMnO4
12 g 2,4-Dinitrophenylhydrazine
10 g K2CO3
60 mL H2SO4
2.5 mL 5% NaOH
80 mL H2O
150 mL H2O
200 mL 95% EtOH
Vanilline
Iodoplatinate
3 g Vanilline
10 mL 5% PtCl2 solution
50 mL 95% EtOH
5 mL conc. HCl
15 mL 2 M H2SO4
240 mL 2% aq. iodine solution
Jones
Cerium(IV) Sulphate (Ceric sulfate)
20 g Na2CrO7
10% Cerium(IV) Sulphate
160 mL H2SO4
15% H2SO4, H2O
240 mL water
Morin Hydrate .1% wt % in methanol
Cerium Molybdate (Hanessian's Stain)
Ceric Ammonium Molybdate
5.0 g CeSO4
40 mL H2SO4
25.0 g (NH4)Mo7O24․4H2O
360 mL H2O
450 mLl H2O
10 g Ammonium molybdate
50 mL H2SO4
4 g Ceric ammonium Sulfate