تست نین هیدرین

آزمایش نین هیدرین :

پرولین + نین هیدرین← زرد

روش کار:

1-2 سی سی محلول اسید آمینه + چند قطره محلول نین هیدرین  ← 5 ذقیقه در بن ماری جوش

با آب مقطر نیز Blank می گذاریم.

همه آمینواسیدها جواب مثبت می دهند ← ارغوانی

پرولین  ← زرد

آب مقطر  ←  بی رنگ

Procedure

Add about 2 mg of the sample to 1 mL of a solution of 0.2 g of ninhydrin (1,2,3indanetrione monohydrate) in 50 mL of water. The test mixture is heated to boiling for 15-20 sec; This reaction is important not only because it is a qualitative test, but also because it is the source of the absorbing material that can be measured quantitatively by an automatic amino acid analyzer. This color reaction is also used to detect the presence and position of amino acids after paper chromatographic separation.

Positive Test

A blue to blue-violet color is given by a-amino acids and constitutes a positive test. Other colors (yellow, orange, red) are negative.

Complications

Proline, hydroxyproline, and 2-, 3-, and 4-aminobenzoic acids fail to give a blue color but produce a yellow color instead.

Ammonium salts give a positive test. Some amines, such as aniline, yield orange to red colors, which is a negative test

physical propertis of nin hydrin

 Physical State: Crystalline powder
Appearance: slightly yellow
Odor: characteristic odor
pH: Not available.
Vapor Pressure: Negligible.
Vapor Density: 6.16 (air=1)
Evaporation Rate:Not applicable.
Viscosity: Not available.
Boiling Point: Not available.
Freezing/Melting Point:466 deg F (dec)
Autoignition Temperature: Not applicable.
Flash Point: Not applicable.
Decomposition Temperature:466 deg F
NFPA Rating: (estimated) Health: 2; Flammability: 1; Reactivity: 0
Explosion Limits, Lower:Not available.
Upper: Not available.
Solubility: Soluble in water.
Specific Gravity/Density:0.86
Molecular Formula:C9H6O4
Molecular Weight:178.14