Operation Research 2 Phase Technique - PowerPoint

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					Research Institute Facilities
By Christine Andrews, Karen Gogala & Marja Simpson
    Horticulture Centre Equipment

•   Laboratory facilities with large working space
•   Trial Site Area
•   Plant Growth Cabinets
•   Large Capacity Dehydrating Oven
•   Cool Room
•   Video and scanner based image analysis system
•   Glasshouses with automatic heating and cooling
•   Steam generator
•   Automatic weather stations
Horticulture Centre Laboratory
          & Trial Site
          Plant Growth Cabinets
•   1 large cabinet, 2 smaller cabinets
•   Temperature and humidity controlled
•   Lighting intensity variable by switching lamps
•   24 h timers provide control between instruments
  Dehydrating Oven & Cool Room




Operating range is +10oC to + 200oC
Video and scanner based image analysis system
    •  Captures images with coloured video camera, Delta-T SCAN splash-
    protected flatbed scanner
    •   WinDIAS and Delta-T SCAN image analysis software analyse
    images
    •   Usage: WinDIAS- Measurement of the area of healthy and
    diseased plant leaves
             Delta-T SCAN- Leaf measurement, Root length
    measurement, object size analysis, eg. soil particles, seeds,
    measurements from photographs or copies, count objects, eg. seeds
                     Glasshouses
•   Glasshouse size 3m x 7.5m x 3m
•   Automatic cooling and heating system
•   Winter heating capacity 10oC overnight, 20oC day
•   Summer cooling 20-25oC
•   Lighting is provided by incandescent and fluorescent
    lamps which is 24 h timer controlled
          Steam Generator
• Soil and plastic container treatment to control
soil borne fungal diseases, nematodes and weeds
   Automatic Weather Station
• 2 stations
• Records: Wind speed, wind direction, air temperature,
  rainfall, relative humidity, solar radiation, logger
  calculates evaporation
• Data available online
  http://www.orange.usyd.edu.au/research/weather/index.htm
                 Soil Shaker
• The Endecotts EFL 2000 is a vibrating shaker that is
  used to carry out sieve tests in conjunction with sieve
  stacks for particle sizing of various material samples.
• Sizes of sieves available:        1.0mm
                                    2.0mm
                                    500micron
                                    250micron
                                    125micron
                                    63micron
    Equipment – Research Lab

•   Atomic Absorption Spectrometer
•   UV/Visible Spectrometer
•   Scanning Electron Microscope
•   Fluorescence Microscope
•   PCR System
•   Automontage Microscope
•   GIS System
•   GC/MS
•   HPLC
        Atomic Absorption
       Spectrometer (AAS)

• Measures the amount
  of light absorbed by
  atoms
• Liquid sample
  aspirated, aerosolized
  & mixed with gas
• Ignited in flame
• Atoms reduced to free
  state which absorbs
  light
UV/VIS Spectrophotometer

• Measures amount of
  light a sample absorbs
• A beam of light passes
  through onto a detector
• Amount of molecules in
  a sample can be
  detected
• Both UV & visible
  spectra
      Scanning Electron
         Microscopy
• Creates magnified
  images by using
  electrons instead
  of light waves
• Shows 3D images
  at much higher
  magnification
• Samples prepared
  – sputter coater
 Fluorescence Microscopy

• Sample you want to study
  is the light source
• Energy absorbed by atom;
  it gets excited
• Electron jumps to a higher
  energy level
• Drops back to ground
  state, emits a photon
  (fluorescing)
         PCR Room
• Polymerase Chain
  Reaction is a
  molecular biological
  technique for
  amplifying DNA without
  using a living
  organism.

• PCR is commonly used
  in medical and
  biological research
  labs for a variety of
  tasks.
Automontage
 Microscopy
• Perfectly focused
  3D images
• Increased depth
  of field software
• Allows images of
  small insects
  almost as good
  as the specimen
  itself
GIS System
• Manages spatial
  data and
  associated
  attributes.
• It is a computer
  system capable of
  integrating, storing,
  editing, analysing,
  and displaying
  geographically-
  referenced
  information.
HPLC and GC/MS
• High Performance Liquid
  Chromatograph
• Gas Chromatograph coupled with a
  Mass Spectrometer
High Performance Liquid
Chromatograph
•   Chromatography—what is it?
•   Liquid Chromatography
•   Basic Operation
•   Equipment used
•   Types of Chromatography
•   Applications for HPLC
What is chromatography?
• Chromatography –’colour’ and ‘to write”
• Originally described by Tswett in 1906 who devised a
  method to separate plant pigments using a tube filled with
  CaCO3.
• Basically it is a broad range of physical methods used to
  separate and /or to analyse complex mixtures
• Components to be separated are distributed between two
  phases:a stationary phase bed and a mobile phase which
  flows through the stationary bed.
• Individual species are retained by the stationary phase
  (packing) based on various interactions such as surface
  adsorption, relative solubility of the sample in the mobile
  phase and charge.
Chromatography
• LC-mobile phase is a solvent and
  stationary phase is a packed bed of
  silica particles.
Liquid Chromatography
• HPLC is this process conducted at a high velocity and
  under pressure.
• Sample can be in an aqueous form or in an
  organic/aqueous form.
• Sample is injected onto the column and is pushed through
  by the mobile phase(eluent) under high pressure.
• Components are retained and separated on the
  column.They elute at different times depending on their
  chemical interaction with the packing in the column.
• The time at which they elute (retention time) is a
  characteristic of that compound.
• After compounds elute,they enter a detector(PDA) which
  creates an electronic signal. The greater the concentration
  of the compound, the greater the signal.
Liquid Chromatography
• Chromatogram
Basic Operation
Equipment used
• Shimadzu HPLC
Types of chromatography

  • Adsorption




  •Ion Exchange
     Applications
•   Chemical separations
•   Identification
•   Quantification
•   Purification
•   Cosmetics,energy,food,life sciences
    pharmaceutical, biomedical, drug detection and
    identification.




                                 Carbohydrates in vegetables
           Herbicides
Gas Chromatograph/Mass
Spectroscopy (GC/MS)
• Gas Chromatograph
• Mass Spectrometer
Gas Chromatograph
• Mobile phase is an inert gas such as helium
• Sample is injected into a heated injection port, becomes
  vapourised and travels onto the column by means of the
  carrier gas.
• Column is made of fused silica onto which is coated the
  liquid stationary phase and it is enclosed in a heated
  zone(oven)



• Compounds become separated as they interact with the
  column
• Variables are temperature,gas flow, and column
  specifications.
• Separated compounds identified by specific
  detector.(FID.NPD,ECD)
Gas Chromatography
• Schematic diagram
Mass Spectrometer
• Creates charged particles (ions) from
  molecules.
• Analyzes those ions to provide
  information about the MW of the
  compound and it’s chemical structure
• Many types of MS which allow wide range
  of analyses.
• GC/MS is the coupling of GC with MS
GC/MS



•  A. Capillary column interface which connects GC to
  mass spectrometer
• B. Sample enters ionization chamber
• Ionization occurs. A beam of electrons impacts the sample
  molecules which lose an electron becoming positive (M+)
• C. A positive potential is applied to repel the + ions out of
  the ionization chamber and into the mass analyser.(filter)
GC/MS
• Mass analyser separates the positively charged particles
  under vacuum according to their mass.




• Particles then enter a detector which sends information to
  the computer and resulting chromatograms give a mass
  spectrum of the sample components.
• Identification of the compound relies on the fact that every
  compound has a unique fragmentation pattern.
GC/MS
• Mass Spectrum Jamaican coffee
Shimadzu GC/MS

				
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