Chem Exam 1 Flashcards
What are the approximate wavelengths of visible light?
400-700 nm
How does wavelength relate to energy?
Shorter wavelength = higher energy
Longer wavelength = lower energy
What is the formula that relates absorbance and transmittance?
A = -log T = -log (T1/T0) = 2- log %T
Examples of light sources for spectrophotometer
tungsten lamp, quartz-halogen lamp, deuterium discharge lamp, mercury arc lamp, or laser
Types of monochromators
Colored glass filters, prism, interference filters (mirrors), diffraction grating
What are these types of: colored glass filters, prisms, diffraction grating
Monochromators
What is the measure of the monochromator’s spectral purity?
Bandwidth - specifically a narrow bandwidth
What characteristics must a cuvette have in spectrophotometry?
Standard in size, clean, unscratched
Types of photodetectors
Photocell, phototube, photomultiplier tube, diode array
Advantages and disadvantages of a photocell as a photodetector
Adv: cheap, easy, does not need a current/voltage
Disadv: needs a LOT of light, quick to fatigue
Advantages and disadvantages of a phototube as a photodetector
Adv: simple to use
Disadv: requires a current, requires a lot of light, cannot multiply a light signal, quick to fatigue, requires time between measurements
Advantages and disadvantages of a photomultiplier tube
Adv: most sensitive of the detectors, can multiply a low light signal, slow to fatigue
Disadv: expensive and very complex
Advantages and disadvantages of a diode array
Adv: can measure more than one wavelength simultaneously, specific
Disadv: not quite as sensitive as photomultiplier tube
What are some measurements used to assure quality results from a spectrophotomer?
Linearity, stray light check, wavelength check
How can we limit background interference in spectrophotometry?
Sample blanking - contains everything except patient sample to detect any baseline absorption
What are the light scattering measurements?
Nephelometry, turbidimetry, laser light scatter (flow cytometry)
Nephelometry
Nephelometry measures the scatter of light from particles in a reaction solution with a detector for scattered light at an angle to the source light (forward scatter position)
Turbidimetry
Measures the reduction in amount of light by particles in a reaction solution, can be measured with most traditional spectrophotometers
What is unique about the light source for A.A.S.?
Lamp contains the material that will be measured (heavy metals)
What does a chopper do?
Chops the light signal coming from the lamp to allow for detector to distinguish between light coming from lamp and light coming from excited atoms in the flame
What does the monochromator do in A.A.S.?
Selects wavelength that reaches the detector after light enters sample
Protects the detector from large amounts of light
What does the detector do?
Distinguishes between light from the light source and light emitted by excited atoms in the flame
Name a few elements A.A.S. measures
Calcium, lead, copper, chromium, aluminum
Advantages of A.A.S.
sensitive and precise, can be used for analytes that are difficult to measure in other ways (measures heavy metals)
Disadvantages of A.A.S.
possibility for chemical interference and possibility of excitation of atoms by flame if flame temp is too high
need a separate lamp for each analyte
What does lanthanum do?
acts as a competitor for phosphate leaving calcium free for measurement during A.A.S.
Limitations in calcium measurement? What is used to help with this?
calcium will frequently bind to phosphate and calcium levels will not be accurately measured. Lanthanum is added to act as a competitor for phosphate.
What is a fluorophore?
A substance that can fluoresce –> it absorbs energy at higher energy wavelengths and emits light at lower energy wavelength