Instrumentation

Almira Coleen · 28問 · 1年前

通報

問題一覧

Instrumentation

Spectrometry, Luminescence, Electroanalytic Methods, Chromatography

Measuring of interaction with light and matter

Spectrometry

Measures absorption of light

Spectrophotometry

Measuring of light which needs atoms ratio

Atomic Absorption

Measures mass charge ratio of ions

Mass Spectrometry

Emission of light by a substance after it absorbs energy

Luminescence

Emission of light because of chemical reaction

Chemiluminescence

Involves in measurement of-electro electrical properties

Electrophoresis

Involve in measuring potential difference - used for ISC or Iso-electrical focusing - Electrolytes, Sodium, Potassium

Potentiometry

Lead is the current flow to the solution and it uses potential different charge

Amperometry

Based on separation of components, mobile -> gas, liquid and stationary -> thin layer

Chromatography

• Measures light energy • Involves in measuring of light • Based on the property -> colored solutions • Absorb light in a specific wavelength

Spectrophotometer

• Different types of Radiation • 400nm to 700nm • Gamma rays, X-rays, Infrared, Radio waves

Electromagnetic Spectro

True or False Increased in wavelength, Decrease in energy

True

True or False Decrease in energy, Increase in Wavelength

True

In Radio waves, it is the strongest energy

Violet

In Radio waves, it is the lowest energy

Red

Concentration: Directly proportional to the amount of light absorbed and to the amount of transmitted light.

Beers Law of Spectrophotometry

True/ False Increase in substance, Decrease in transmit of light

True

Parts of Beers Law Concentration: • A = E x b x c • A = • E= • b = • c =

Absorbance, Epsilon -> molar absorbidity, length of light path, concentration of substance/absorbance

• Measures the light transmitted by the solution • Determine the concentration of the substance in the solution

Beer's Law

Components of Spectrophotometer

Light source, Entrance slit, Monochromator, Exit slit, Sample cell, Photo detector

Provide incident light for the system

Light Source

Visible and clear infrared spectrum

Incandescent Tungsten/ Tungsten Iodide Lamp

Use for UV spectrum

Deuterium Discharge Lamp and Mercury Arc Lamp

Infrared Spectrum

Silicone Carbide

Isolates specific wavelength from the light source

Monochromator

Colored Glass Filters, Interference Filter, Prism, Diffraction Grating

Inexpensive but has low transmittance of wavelength

Colored Glass Filter

• Constructive interference of waves • More precise than color glass filter • Produce very narrow waves

Interference Filter

Separates white light into a continuous spectra

Prism

• Widely used monochromator • Separates light into component wavelength as wavelength bends as they pass a sharp corner

Diffraction Grating

Exclude unwanted or stray light

Entrance Slit

Controls the width of the light beam

Exit Slit

• Cuvette or Analytical Cell • Holds the solution of which the absorption is to be measured

Sample Cell

Types of Sample Cell

Glass Cuvette, Quartz or fused silica

A sample cell which is visible for visible range

Glass Cuvette

Sample Cell used for UV range

Quartz or Focused silica

Reminder! In using cuvette make sure that it is free from _______, it will scatter the light ----> NADH

Scratches

Converts transmitted radiant energy into an equivalent amount of electrical energy

Photo Detector

Photo detector

Photo Cell, Photo Tube, Photo multiplier Tube, Phototransistor & Protoiodide

A barrier layer cell, selenide cell Generates electromotive force No external voltage Output is not amplified

Photocell

Similar to the other one but requires external voltage

Phototube

Amplifies radiant energy (200x sensitive)

Photomultiplier Tube

Uses a photosensitive positive-negative junction diode to produce a photo current

Photodiode

A moving needle on a dial or a digital display which indicates the amount of light passing through a sample.

Read-out Device

Measures the light transmitted by a solution to determine concentration of a substance in the solution

Spectrophotometry

________ Cuvettes are preferably used because they reduce incident light

Rectangular

Number one source of error in spectrophotometer is cuvette _______.

Mismatch

To check the WL calibration of spectrophotometer, ______ or ______ filters are used.

Holmium Oxide, Didymium Filters

Principle: Absorbance is measured at two different WL using a light from either monochromators that would analyze samples and reference cuvette.

Double Beam Spectrophotometer

Two types of Double Beam Spectrophotometer

Double Beam in Space, Double Beam in Time

Passes light in two directions. One beam is directed to sample and the other is to reference cuvette simultaneously.

Double Beam in Space

Splits light using a rotating chopper. One beam is directed to sample and the other is for reference cuvette with an interval time.

Double Beam in Time

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