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Evaluation of the
Chlorophyll Content of Different "Greens"
by Filter-Fluorometry


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INTRODUCTION:

      There are many WAVELENGTHS of Energy in the Electromagnetic Spectrum; from the ultra-short region of X-Rays to the very long wavelengths of Microwaves. In between is a spectral region of Light called the UV-VIS (Ultra-Violet / Visible) Spectrum. These wavelengths of energy from 190nm. beginning in the UV, through 900nm. at the end of the VIS, can generate a tremendous amount of descriptive information about molecules that ABSORB certain portions of this Spectrum.
      Most Organic molecules will simply ABSORB some of this light energy, and generate a Peak on a Spectrum. There are a few special Molecules that will undergo a different type of Light ABSORPTION, and will GIVE OFF Light of a lower energy relative to the wavelength of Light they are exposed to. This is the nature of FLUORESCENCE. Fluorometric instruments are designed so that the Light SOURCE and the DETECTOR are offset (angled) by 90o, so the only energy that can reach the Detector must be given off by the Sample itself.

PURPOSE:

      Natural, Plant-based CHLOROPHYLL is a 4-ring structure coordinated with some "METAL" Ion… usually Magnesium, but it can also be Copper or Manganese.
      Because of its complex molecular structure, Chlorophyll is also FLUORESCENT, and has a pale Yellow-Green response when exposed to mid-range UV light. BY running a "REFERENCE" Plant, such as Spinach, we can compare either different Varieties of "GREEN" vegetables, or design a Greenhouse Experiment to see the effect of Environmental variables on the "quality" of the Chlorophyll in the Plant.

FUN-SCI Kit: Includes all supplies Buy it!

INSTRUMENTS NEEDED:

  • Model Mini FL-900 / 380 Fluorometer w/ Fluorescent Filter set (from BUCK Scientific) Buy it!
  • Filter Set for EXCITATION at 390 & 460nm. and EMISSION Response range of 540-700nm. Buy it!
  • 10mm. square Quartz Fluorescence Cuvettes (clear on ALL 4 sides, unlike regular Spectro cuvettes) Buy it!

    SUPPLIES:
  1. Disposable GLASS Vials (to hold Sample Extracts)
  2. Assorted GREEN Plants (Germanium Leaves, Grass, Alfalfa, etc) OR
  3. GREEN Vegetables (Spinach, Kale, Mustard Greens, Collard Greens, Turnip Greens, Flat & Curly Parsley, etc).
  4. Extracting Solvents: Methanol, Hexane, Methylene Chloride, Acetone
  5. Reagents for pH adjustment: 0.05N Hydrochloric Acid, 0.05N Sodium Hydroxide
  6. Reagents for Red-Ox adjustments: 3% Hydrogen Peroxide, 0.1% Sodium Bisulfite
  7. Disposable plastic pipets / eyedroppers
  8. Disposable 100ml. Beakers
  9. Mortar & Pestle (agate or porcelain)
SAMPLES:
  • Student can bring in other TYPES of Plants or Vegetable GREENS from the supermarket or farm stand
  • Download this experiment
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