PHARM.

ANALYSIS 2 LEC-MODULE 1

Saffia A. Abu-shendi
 §Photometry is the science
of measuring visible light in
units that are weighted
LESSON 2: according to the sensitivity
SPECTROMETRIC of the human eye.
CALCULATION §This measurement is used
to determine the amount of
an analyte in a solution
or liquid.
§ Is the measurement of light.
§ It is the most commonly used measurement in the
laboratory because light can undergo
§ Absorption – spectrophotometry
§ Scatter – nephelometry
§ Reflection – turbidimetry
§ Emission – flame spectroscopy
§ Fluorescence – fluorometry
§ Light is a form of electromagnetic energy that appears to
travel in waves.
§ The wavelength or distance between peaks of light waves is a
function of its energy.
§ As the wavelength is increased, emitted energy is decreased.
§ As the wavelength is decreased, emitted energy is increased.
§ The color of light is a function of its wavelength.
§ A law stating that the power of a transmitted radiant
beam decreases exponentially as the concentration
of the solution containing the absorbing chemical
species increases arithmetically.
§ According to this
law, the amount
of light absorbed
is directly
proportional to
the
concentration of
solute in the
solution under
analysis.
§ Where:
§ A = absorbance
§ a = absorptivity coefficient
(constant)
§ b = length of light path
(constant)
§ C = concentration
§ The term “absorbance” is used to represent the
logarithm of transmittance.
§ Absorbance increases linearly with concentration.
§ Absorbance is also known as “optical activity”,
“absorbancy” or “extinction coefficient”.
§ In spectrophotometric analysis, the absorbance of an
unknown concentration (u) of a particular constituent
is compared with the absorbance of a known
concentration (s).
§Au / As = Cu / Cs
§ It states that the power of a transmitted radiant beam
decreases exponentially as the thickness of the
solution containing the absorbing chemical species
increases arithmetically.
§ Thickness is inversely proportional to transmittance
and directly proportional to absorbance.
§ A combination of the above laws and relates the
power of the incident beam and transmitted beam to
the thickness and concentration of the solution
containing the absorbing chemical species.
§ Calculation step:
§1-Determine the concentration of active constituent (Cu) in the final dilution.
§ Cu = Au/As x Cs

§2- Determine the concentration of the sample (U) taken in the final dilution.
§ U = wt/mL x # of dilutions

§ 3-Determine the quantity of the active constituent in a given quantity of

sample.
§ Amount/tab = Cu/U x Average weight of tablet

§ 4- Compute for the % labeled amount.

§ % LA = Amount per tablet x 100%
§ Labeled claim
§ Calculation step:
§1-Determine the concentration of reference standard (Cs) in the final dilution.
§ Cs = wt/mL x # dilutions

§2-Determine the concentration of trioxalen (Cu) in the final dilution using the
spectrometric equation
§ Cu = Au/As x Cs

§ 3- Determine the concentration of sample present in the final dilution.

§ U = wt/mL x # dilutions

§ 4- Determine the % purity.

§ % purity = Cu/U x 100%

§ Short- Method : % Purity = Au/As x S/U x 100%