Vitamins Laboratory: Reactions for Vitamin A, B1, B2, B6, B12, C, and Folic Acid

Vitamins Implementation of Practicum a. Tools and materials Vitamin compound reagents, test tubes, dropper pipettes, tube racks, glass objects, cover glasses, microscopes, spirit lamps, sublimation rings b. Work procedures 1. Vitamin A a. Fluorescence: faded yellow green b. Substance + AgNO3→ rosa c. Substances in water → orange d. Substance + Phosphomolybdate → blue e. Carr - Price reaction: Substance in 10 drops of chloroform + 2 drops of anhydrous acetic acid + SbCl3(slightly) → blue → red brown. f. Trichloroacetate (TCA) reaction: Substance + 1 ml trichloroacetate in chloroform, shake → greenish blue. 2. Vitamin B1 a. Formaldehyde Azo test: Substance + azo benzene + H2SO4 + NaOH+ formaldehyde→ red b. Substance + HgCl2→↓ white c. Substance + 10% Pb-acetate solution and 1 mL NaOH 6 N→ yellow color, heat → ↓ brown-black d. Substance + 10 drops of bismuth nitrate solution + 2 drops of 5% KI solution → ↓ orange red e. Substance + NaOH + K3Fe(CN)6+ amyl alcohol → when shaken, it fluoresces purple blue on the amyl alcohol layer f. Zat + Nessler → black yellow g. Substance + NaOH→ yellow green + KMnO4→ green h. Substance + NaOH, heat → yellow i. Substance + ninhydrin → stable yellow j. Substance + Phosphomolybdate → blue k.Crystal Reaction: fe. Complex, picric acid, Bouchardat. 3. Vitamin B2 (Riboflavin) a. Substance + H2SO4deep→ cherry red/orange red b. Substance + HNO3solid → yellow green c. Substance + concentrated HCl → yellow green d. Substance + concentrated acetic acid → cloudy yellow e. Zat + Frohde→ red f. Substance + Liebermann Bouchard→greenish yellow (-) g. Substance + Salkowski→red (+) h. Substance + Cuprifil→ (+) i. Substance + Marquis→ orange + gas j. Substance + AgNO3→cherry red, or over time a red precipitate forms k. Substance + KMnO4→purple red (+) l. Substance + Diazo A + Diazo B + NaOH→ yellow → orange red m. Substance + FeCl3→ orange red/brownish red n. Substance + Diazo → red o. Zat + Fehling → ↓red p. Substance + Nessler → ↓ q. Substance + water → solution+ AgNO3→color change occurs. Some time can form ↓red r. Substance in water → yellow ± orange in fluorescent light yellow dark green + NaOH 3N/HCl 3 N, fluorescence disappears s. Crystal reactions: Dragendorf, Bouchardat, Mayer, Picric acid, Sublimation 4. Vitamin B6 a. Formaldehyde Azo test: Substance + azo benzene + H2SO4 + NaOH+ formaldehyde→ red b. Substance + Diazo A + Diazo B + NaOH ad base - yellow orange red c. Substance + FeCl3→ orange to dark red/blood red d. Substance + 2 drops of CuSO solution42% and 10 drops of NaOH 3 N → blue-purple e. Zat + Nessler → black yellow f. Substance + NaOH→ yellow green + KMnO4→ stable yellow green g. Substance + ninhydrin → stable yellow h. Substance + Phosphomolybdate → blue i. Test substance + AgNO3→ precipitate + NH4OH → soluble precipitate j. Substance + Diazo A & B + NaOH → yellow → orange red k.Crystal Reaction: Fe. Complex, picric acid, Bouchardat. 5. Vitamin B12 a. Substance + Nessler → black b. Substance + KMnO4, in cold conditions → purple color is reduced to black c. Substance + KNO3in acid → Yellow precipitate (Co identification) 6. Vitamin C a. Substance + Nessler → black b. Substance + KMnO4, in cold conditions → purple color is reduced to black c. Substance + AgNO3,→ gray precipitate, heat→ silver mirror d. Substance + NaHCO35%+ 2 drops of FeCl solution3→ red purple e. Substance + Phosphomolybdate → purple f. Substance + FeCl3→ negative g. Substance + ninhydrin → negative h. Cuprifil, (Substance + NaOH ad alkalis + CuSO41% → purple blue) i. Test substance + NaOH + FeSO4(liquid) → violet green color j. Substance + NaOH → yellow + KMnO4→ green k. Substance + NaOH, heat → rosa l. Substance + HNO3+ AgNO3→ ↓grey m. Reducing Fehling's reagent, (Substance + Fehling's reagent A: Fehling's B (1:1) + NaOH 2N ad alkalis, heat in WB → ↓ Cu2O (brick red)) n. Reducing Luff's reagent, (Substance + Luff's reagent, heat in WB → ↓ Cu2O (brick red)) o. Reducing Barfoed's reagent (3 ml of Barfoed's reagent + 1 ml of sample solution, heat in WB → ↓ Cu2O (brick red)) p. Substance + 15 drops of Benedict's reagent, heat until boiling 2 minutes → ↓ yellowish green to red q. Substance + water + NaHCO3(solid)+ FeSO4(solid), shake, let stand → purple+ H2SO4 dilute → purple color disappears r. Substance in water + Potassium Cupril Tartrate → ↓ orange, heated → red brown 7. Folic Acid a. Solution in Na2CO3yellow b. The solution in hot dilute HCl is yellow. If you add H2SO4 the thick yellow color is gone c. The solution in NaOH fluoresces blue green d. The substance in heated nitric acid + NaOH→ yellowish green fluorescence e. Substance + Marquis reagent → red f. Substance + H2SO4→ green, after a while it disappears