• Fume Hood
• Nebulizer - 250 ml, 20/40 Joint
  Mfg - Kontes
  Catalog # - K4225300250
  Mfg - Chemglass
  Catalog # - NC9755621
• Stirring hot plate
• Magnetic stir bars
• Assorted glassware
• Molybdenum VI Oxide - Aldrich Catalog Number - 22,181-3
• Molybdenum metal - Aldrich Catalog Number - 20,382-3
• Sulfuric acid
• Deionized water

Procedure:
Preparation of Reagent 1

• Dissolve 40.1g of molybdic anhydride in 1L of conc. sulfuric acid with heat and stirring overnight.
• Cover the beaker or flask with plastic film (Saran wrap) and secure with rubber band to prevent uptake of moisture.
• Produces a clear, light blue solution.

Preparation of Reagent 2

• Dissolve 1.78g of molybdenum metal using 500mL of Reagent 1 with heat and stirring overnight.
• Cover the beaker or flask with plastic film (Saran wrap) and secure with rubber band to prevent uptake of moisture.
• Produces a dark blue solution.

Preparation of Phosphorus Spray

• To 120mL of deionized water add with stirring: 40mL Reagent 1 40mL Reagent 2
• Use extreme caution while adding reagents to water.
• Important! Always add reagents to water, not water to reagents. Adding water to sulfuric acid solutions can be explosive.
• Thoroughly mix the solution, allow to cool, and pour into a nebulizer.

Application of Phosphorus Spray

• In a fume hood, spray TLC plates with molybdate solution until plate is covered with reagent.
• Heat plate slightly until spots completely develop.
• Phosphorus containing compounds will appear as blue spots on white background. Additional heating will char unsaturated compounds.

Reference

• Ellingson, J.S. & William E.M. Lands (1968) "Phospholipid Reactivation of Plasmalogen Metabolism", Lipids, pp. 111-120.

• Nebulizer
• Assorted glassware
• n-Butanol
• Ninhydrin reagent - Acros Catalog Number - 41572-0250
• Deionized water

Procedure:
Preparation of Ninhydrin Spray

• Saturate n-butanol with water by shaking 200mL of n-butanol with 200mL of deionized water in a separatory funnel. Discard the water phase (lower phase).
• Dissolve 200mg of ninhydrin reagent in saturated n-butanol with stirring.
• Produces a clear, light yellow solution.
• Thoroughly mix the solution and pour into a nebulizer.

Application of Ninhydrin Spray

• In a fume hood, spray TLC plates with ninhydrin solution until plate is covered with reagent.
• Heat plate until spots completely develop.
• Primary amine containing compounds will appear as pink spots on white background.

Reference

• Skipski, V.P., Peterson, R.F., & M. Barclay (1962) J. Lipid Res. 3:467.