Specto poto meter ko

Ir Rudy Gunung Sugih

7.1 Using the Spectrophotometer to Detect Molecules

Molecules are too tiny to be seen.

When an indicator solution changes color, that means a
molecule of interest is present.
A VIS spectrophotometer uses white light, composed of the
visible spectrum, to detect colorful molecules.
Absorbance, Transmittance, and Reflection. A spectrophotometer measures
how light interacts with atoms or molecules in a sample.
Parts of a Spectrophotometer
• Lamp
• Prism
• Sample holder
• Display

How a Spectrophotometer Works

White light hits grating or prism

Light is split into colors of the rainbow
Wavelength knob directs different colors toward sample
 Colors of Light in the Visible
Spectrum. Humans can see light
with wavelengths of about 350 to
700 nm.

How a UV Spectrophotometer Works. Similar to a

VIS spectrophotometer, the UV spec shines
ultraviolet light or visible light on a sample, and a
detector measures the amount of light that passes
through, or is absorbed by, the sample.
How Concentration Affects Absorbance. If a sample has twice as many molecules as
another, it can absorb twice as much light. This is true at any wavelength. It is important to
know a sample’s wavelength of maximum light absorbance, so that the difference in
absorbance due to concentration is obvious.
 Vocabulary

• Ultraviolet light – a wavelength of light that is used to detect colorless molecules

• Spectrophotometer – an instrument that measures the amount of light that passes through (is
transmitted through) a sample
• Nanometer – 10-9 meters; the standard unit used for measuring light
• Visible light spectrum – the range of wavelengths of light that humans can see, from
approximately 350 to 700 nm; also called white light
• Transmittance – the passing of light through a sample
• Absorbance – the amount of light absorbed by a sample (the amount of light that does not pass
through or reflect off a sample)
• Tungsten lamp – a lamp, used for VIS spectrophotometers, that produces white light (350 –
700 nm)
• Deuterium lamp – a special lamp used for UV spectrophotometers that produces light in the
ultraviolet (UV light) part of the spectrum (200 – 350 nm)
• % transmittance – the manner in which a spectrophotometer reports the amount of light that
passes through a sample
• Absorbance units – (abbreviated “au”) a unit of light absorbance determined by the decrease
in the amount of light in a light beam
• Absorbance spectrum – a graph of a sample’s absorbance at different wavelengths
• Lambdamax – the wavelength that gives the highest absorbance value for a sample
A pH meter is used to adjust the pH of solutions or to watch the pH of a solution change over time. The
pH electrode is sitting in a 50-mL tube of storage solution.
 Vocabulary

• Acid – a solution that has a pH less than 7

• Base – a solution that has a pH greater than 7
• Hydrogen – a hydrogen atom which has lost an electron (H+)
• Neutral – uncharged
• Pepsin – an enzyme, found in gastric juice, that works to break down food (protein) in
the stomach
• pH paper – a piece of paper that has one or more chemical indicators on it and that
changes colors depending on the amount of H+ ions in a solution
• pH meter – an instrument that uses an electrode to detect the pH of a solution
• Buffer standards – solutions, each of specific pH, used to calibrate a pH meter
7.3 Buffers

Buffer solution resists changes in pH.