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Calibration of

Performing at least the following checks should
validate instrument function:-

Wavelength accuracy
Wavelength accuracy means that the
indicated on the control dial is the actual
of light passed by the monochromator.
It is most commonly checked using standard
absorbing solutions or filters with absorbance
maxima of known wavelength.
Didymium or holmium oxide in glass is stable
The filter is placed in the light path and the
wavelength control is set at the wavelength at
which maximal absorbance is expected.
The wavelength control is then rotated in either
direction to locate the actual wavelength that has
maximal absorbance.
If these two wavelengths do not match, the
Stray light
optics must be adjusted to calibrate the
monochromator correctly.
Any wavelengths outside the band transmitted
by the monochromator.
The most common causes of stray light are
reflection of light from scratches on optical
surfaces or from dust particles anywhere in the
light path and higher-order spectra produced by
To check for stray light in the near-UV region, for
example, insert a filter that does not transmit in
the region of 200 nm to 400 nm.
If the instrument reading is greater than 0% T,
stray light is present.
Certain liquids, such as NiSO4, NaNO2, and
acetone, absorb strongly at short wavelengths
and can be used in the same way to detect stray
light in the UV range.
Linearity is demonstrated when a change in
concentration results in a straight-line calibration
curve, as discussed under Beers law.
Colored solutions may be carefully diluted and
used to check linearity, using the wavelength of
maximal absorbance for that color.
Sealed sets of different colors and
concentrations are available commercially.
They should be labeled with expected
absorbance for a given band pass instrument.
Less than expectedabsorbance is an indication
of stray light or of a band pass that is wider than
Sets of neutraldensity filters to check linearity
over a range of wavelengths are also
commercially available.