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.
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 CoCl2
assists the staining process. The spots are often different colors
against a pink or colorless background.
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.
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.
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.
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.
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.
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.
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.
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.