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Reagen Penampak Noda KLT: P-Anisaldehyde (PAA)
Reagen Penampak Noda KLT: P-Anisaldehyde (PAA)
Reagen Penampak Noda KLT: P-Anisaldehyde (PAA)
Bagi mereka yang bekerja dalam sintesis organik dan ataupun isolasi senyawa-senyawa metabolit
sekunder bahan alam sangat terbantu sekali dengan kromatografi lapis tipis (KLT). Namun demikian
terkadang noda KLT tidak semuanya terlihat dengan lampu uv (254; 366 nm). Maka reagen penampak
noda dibawah sangat bermanfaat.
• 12.5 mL p-anisaldehyde
Requires heating
General purpose • 5 mL AcOH
Range of colors on
p-Anisaldehyde (PAA) (especially • 465 mL EtOH
light pink background
nucleophiles) • 17.5 mL conc. H2SO4 Medium shelf life
To a cold (0 °C) mixture of AcOH (5 mL) and absolute EtOH (465 mL) was added p-anisaldehyde (12.5
mL) followed by slow addition of sulfuric acid (17.5 mL). The resulting clear solution was warmed to rt
and used as is. The excess was stored in the fridge.
This stain is light and oxidation sensitive and will gradually turn pink/orange. The stain should be kept in
an aluminum-covered jar while in use, and the excess should be kept cold and in the dark. Once the stain
turns dark red, it should be discarded and made fresh again.
Difficultly
Chromic acid (K2Cr2O7) • 2.5 g potassium chromate (K2Cr2O7)
stainable
compounds • 100 mL 20% aq. H2SO4
To a cold (0 °C) solution of sulfuric acid (100 mL, 20% v/v aq.) was slowly added potassium chromate
(2.5 g). The resulting clear bright red/orange solution was warmed to rt and used as is. The excess was
stored in the fridge.
• 12 g 2,4-
Requires heating
dinitrophenylhydrazine
Yellow to red spots
2,4-Dinitrophenylhydrazine Aldehydes • 60 mL conc. H2SO4
on light yellow
(DNP) Ketones • 80 mL H2O
background
• 200 mL 95% EtOH
Long shelf life
To a cold (0 °C) mixture of water (80 mL) and absolute EtOH (200 mL) was added 2,4-
dinitrophenylhydrazine (12 g). To the resulting cloudy bright orange solution was slowly added sulfuric
acid (60 mL) resulting in a clear orange solution that was warmed to rt and used as is. The excess was
stored in the fridge.
Solution A:
• 1.7 g Bi5O(OH)9(NO3)4
• 80 mL H2O
Amines • 20 mL AcOH
Dragendorff Organic base Solution B:
• 40 g KI
• 100 mL H2O
5 mL A + 5 mL B in 20 mL AcOH + 70 mL H2O
• 10 g KI
• 1.5 g Bi(NO3)3
Dragendorff-Munier Amines • 20 g Tartaric acid
• 120 mL H2O
• 0.5 g dimethylaminobenzaldehyde
Ehrlich Reagent Amines • 10 mL conc. H2SO4
• 90 mL 95% EtOH
• 1 g FeCl3
• 50 mL MeOH
Ferric chloride Phenols
• 50 mL H2O
• 0.1 g C15H10O7·xH2O
Morin hydrate General Purpose
• 100 mL MeOH
• 0.3 g Ninhydrin
Amino acids
Ninhydrin Amines • 3 mL conc. H2SO4
• 100 mL n-butanol
• 7g
Requires heating
phosphomolybdic
Phosphomolybdic Dark green/black spots on light
General purpose acid
acid (PMA) green background
• 100 mL EtOH
Long shelf life
Phosphomolybdic acid (10 g) was added to cold (0 °C) absolute EtOH (100 g) and then warmed to rt. The
resulting cloudy yellow suspension was filtered over filter paper to give a clear bright yellow solution that
was used as is. The excess was stored in the fridge.
This stain can be kept for months on the bench and in the fridge. PMA stain will gradually turn green but
does not affect the stain.
• 15 g vanillin
Requires heating
General • 250 mL EtOH
Vanillin purpose • 2.5 mL conc. H2SO4
Range of colors
Medium shelf life
To a cold (0 °C) clear colorless solution of vanillin (15 g) in absolute EtOH (250 mL) was slowly added
sulfuric acid (2.5 mL). The resulting clear solution was warmed to rt and used as is. The excess was
stored in the fridge.
This stain is light and oxidation sensitive and will gradually turn dark. The stain should be kept in an
aluminum-covered jar while in use, and the excess should be kept cold and in the dark. Once the stain
turns black, it should be discarded and made fresh again.