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Ninhydrin reagent solution

ID: 198249  |  No.: 653  |  Alfred Krause

School experiment from Year 5 onwards

Category: Evidence

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Permanent archiving (DOI)

DOI: https://doi.org/10.5281/zenodo.22811670

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Procedure

Dissolve 0.2 g of ninhydrin (2,2-dihydroindane-1,3-dione; C₉H₆O₄; 1.1 mmol) in 10 ml of ethanol (98 per cent) in a test tube whilst gently heating.  The pale yellow solution is decanted into a brown glass bottle and can be used immediately for the detection of primary and secondary amines. The solution can be stored in the fridge at 5–7 °C for an extended period.

Spray reagent solution for thin-layer chromatography:
0.05 g of ninhydrin is dissolved in 10 ml of ethanol (96 per cent) and, after the addition of approximately 0.2 ml of acetic essence (25 per cent), is transferred to a brown glass spray bottle. The solution is stored in the fridge. 

Equipment & materials

Test tubes, 50 ml measuring cylinder or glass pipette, disposable plastic pipette, 25 ml amber glass bottle, 10 ml brown spray bottle (10 ml spray bottle (Tip: an empty, cleaned brown nasal spray bottle works well for this))

Hazardous substances
Name Formula CAS no. H-statements P-statements Pictograms
Ninhydrin-Sprühlösung 0,5% in Ethanol C₉H₆O₄ 485-47-2
H225 – Liquid and vapour are highly flammable.
German original: Flüssigkeit und Dampf leicht entzündbar.
H319 – Causes serious eye irritation.
German original: Verursacht schwere Augenreizung.
P210 – Keep away from heat, hot surfaces, sparks, open flames and other sources of ignition. Do not smoke.
German original: Von Hitze, heißen Oberflächen, Funken, offenen Flammen und anderen Zündquellen fernhalten. Nicht rauchen.
P240 – Earth the container and the equipment to be filled.
German original: Behälter und zu befüllende Anlage erden.
P403+P233 – Store the container tightly sealed in a well-ventilated place.
German original: Behälter dicht verschlossen an einem gut belüfteten Ort aufbewahren.
P305+P351+P338 – IF IN EYES: Rinse gently with water for several minutes. Remove any contact lenses if possible. Continue rinsing.
German original: BEI KONTAKT MIT DEN AUGEN: Einige Minuten lang behutsam mit Wasser spülen. Vorhandene Kontaktlinsennach Möglichkeit entfernen. Weiter spülen.
GHS02, GHS07
Essigsäure 25% CH₃COOH·aq
H314 – Causes severe chemical burns to the skin and serious eye damage.
German original: Verursacht schwere Verätzungen der Haut und schwere Augenschäden.
P280 – Wear protective gloves / protective clothing / eye protection / face protection.
German original: Schutzhandschuhe / Schutzkleidung / Augenschutz / Gesichtsschutz tragen.
P363 – Wash contaminated clothing before wearing it again.
German original: Kontaminierte Kleidung vor erneutem Tragen waschen.
P308+P310 – IN THE EVENT OF EXPOSURE or if affected: Call the POISON INFORMATION CENTRE or a doctor immediately.
German original: BEI Exposition oder falls betroffen: Sofort GIFTINFORMATIONSZENTRUM oder Arzt anrufen.
P303+P361+P353 – IF IN CONTACT WITH THE SKIN (or hair): Remove all contaminated clothing immediately. Wash the skin with water or take a shower.
German original: BEI BERÜHRUNG MIT DER HAUT (oder dem Haar):Alle kontaminierten Kleidungsstücke sofort ausziehen. Haut mit Wasser abwaschen/duschen.
P305+P351+P338 – IF IN EYES: Rinse gently with water for several minutes. Remove any contact lenses if possible. Continue rinsing.
German original: BEI KONTAKT MIT DEN AUGEN: Einige Minuten lang behutsam mit Wasser spülen. Vorhandene Kontaktlinsennach Möglichkeit entfernen. Weiter spülen.
GHS05
Ethanol C₂H₅OH 64-17-5
H225 – Liquid and vapour are highly flammable.
German original: Flüssigkeit und Dampf leicht entzündbar.
H319 – Causes serious eye irritation.
German original: Verursacht schwere Augenreizung.
P210 – Keep away from heat, hot surfaces, sparks, open flames and other sources of ignition. Do not smoke.
German original: Von Hitze, heißen Oberflächen, Funken, offenen Flammen und anderen Zündquellen fernhalten. Nicht rauchen.
P233 – Keep the container tightly closed.
German original: Behälter dicht verschlossen halten.
P240 – Earth the container and the equipment to be filled.
German original: Behälter und zu befüllende Anlage erden.
P241 – Use explosion-proof electrical equipment / ventilation systems / lighting /...
German original: Explosionsgeschützte elektrische Betriebsmittel / Lüftungsanlagen / Beleuchtung /... verwenden.
P242 – Use only non-sparking tools.
German original: Nur funkenfreies Werkzeug verwenden.
P305+P351+P338 – IF IT COMES INTO CONTACT WITH THE EYES: Rinse gently with water for a few minutes. Remove any contact lenses if possible. Continue rinsing.
German original: BEI KONTAKT MIT DEN AUGEN: Einige Minuten lang behutsam mit Wasser spülen. Vorhandene Kontaktlinsennach Möglichkeit entfernen. Weiter spülen.
GHS02, GHS07
Ninhydrin C₉H₆O₄ 485-47-2
H302 – Harmful if swallowed.
German original: Gesundheitsschädlich bei Verschlucken.
H315 – Causes skin irritation.
German original: Verursacht Hautreizungen.
H319 – Causes serious eye irritation.
German original: Verursacht schwere Augenreizung.
P301+P312+P330 – IF SWALLOWED: If you feel unwell, call the POISON INFORMATION CENTRE or a doctor. Rinse your mouth.
German original: BEI VERSCHLUCKEN: Bei Unwohlsein GIFTINFORMATIONSZENTRUM oder Arzt anrufen. Mund ausspülen.
P305+P351+P338 – IF IN EYES: Rinse gently with water for several minutes. Remove any contact lenses if possible. Continue rinsing.
German original: BEI KONTAKT MIT DEN AUGEN: Einige Minuten lang behutsam mit Wasser spülen. Vorhandene Kontaktlinsennach Möglichkeit entfernen. Weiter spülen.
GHS07

H/P statements: machine translation, with the German original below (authoritative). Name, formula, CAS no. and pictograms are not translated.

Personal protective equipment
  • Safety goggles must be worn.
    German original
    Eine Gestellschutzbrille ist zu tragen.
  • Disposable nitrile gloves are used as splash protection.
    German original
    Als Spritzschutz dienen Nitril-Einmalhandschuhe.
  • A long, fully fastened laboratory coat (protective gown) must be worn.
    German original
    Ein langer, geschlossener Labormantel (Schutzkittel) ist zu tragen.
Protective measures

Use disposable plastic pipettes to measure out the reagent for the detection reaction. 

Caution: Contact with the ninhydrin solution causes the skin to turn blue or purple.

German original

Für die Dosierung des Reagenzes für die Nachweisreaktion Einwegpipetten aus Kunststoff verwenden. 

Achtung: Kontakt mit der Ninhydrin-Lösung führt zu blau- / violett gefärbter Haut.
⚠ Hazards
  • through skin contact
    German original
    durch Hautkontakt
  • through eye contact
    German original
    durch Augenkontakt
  • Fire hazard
    German original
    Brandgefahr
⚠ Further hazards and notes
  • Unintentional release
    Pupils must maintain a safe distance from the source of danger. To remove the contamination, safety goggles, disposable gloves and, where aerosols may be present, an FFP3-rated filtering half-mask must be worn. Contaminated objects or surfaces must be cleaned or wiped down with a damp cloth immediately and, where necessary, disinfected. Use cellulose paper for wiping and absorbing; this must then be autoclaved and disposed of. Detailed information on species-specific procedures can be found in the data sheets of the GESTIS biological agent database.
    German original
    Schüler und Schülerinnen halten sicheren Abstand zur Gefahrenquelle. Zur Beseitigung der Kontamination sind Schutzbrille, Einmalhandschuhe und bei möglichem Vorhandensein von Aerosolen filtrierende Halbmaske der Schutzstufe FFP3 zu tragen. Kontaminierte Gegenstände oder Oberflächen sofort reinigen bzw. nass aufwischen und gegebenen­falls desinfizieren. Zum Wischen und Aufsaugen Zellstoff verwenden, diesen anschließend autoklavieren und entsorgen. Detaillierte Angaben zur speziesabhängigen Vorgehensweisen sind den Datenblättern der GESTIS-Biostoffdatenbank zu entnehmen.
Substitution Substitution 1
The substitution of hazardous substances, as well as their forms of use and methods of application, has been assessed. This experiment is essential for teaching key learning content and may be carried out provided that the restrictions set out in the experiment instructions are observed and the protective measures specified therein are followed.
There are no known hazardous properties of the substances or other risks that would, in principle, preclude the experiment from being carried out by pupils or teachers. The list of substances in DGUV Information 213-098 at degintu.dguv.de has been taken into account.
Media & figures
Ninhydrin_Reagenz.jpg
Ninhydrin_Reagenz.jpg
Ninhydrin_Sprühreagenz.jpg
Ninhydrin_Sprühreagenz.jpg

Ninhydrin solution and spray reagent

Literature

Experimental procedure: Preparation of a ninhydrin reagent solution (2026), Krause, A. and Klemeyer, H. DOI: 10.5281/zenodo.22811670

Ninhydrin – Wikipedia