spectrophotometric koko

‫(‪)spectrophotometric‬‬
‫االسم ‪ :‬كيرلس عاطف جورجي‬
‫المستوي‪ :‬الرابع‬
‫القسم ‪ :‬كيمياء‬
‫د\ اسامه‬
‫د\ نورهان‬
:INTRODUCTION
UV-VIS spectrophotometry is probably the most useful
tool available for quantitative determinations in diverse
areas. It is due to its versatility, accuracy and sensitivity
Spectrophotometry is any technique that uses light to
measure chemical concentrations. A procedure based on
absorption of visible light is called colorimetry The
spectrophotometer is an optical instrument for
.measuring the intensity of light relative to wavelength
:Aim
Determination of iron content in some
pharmaceutical products using 1,10-
.phenanthroline
:Tools
.UV/VIS spectrophotometer )1
.Cuvettes )2
.Beaker )3
.Measure Flask )4
.Pipette )5
: Chemicals
Ferrous ammonium sulfate hexahydrate[Fe(NH4)2.H₂O] •
phenanthorline-1.10 •
Ascorbic acid •
Sodium acetate •
Glacial acetic acid •
.Iron table or solution (unknown) Ex: hydroferrin drop •
: Step
.Preparation of stock solutions
mL standard ferrous solution 10 ppm: weigh a 100 .1
suitable amount of analytical grade
ferrous ammonium sulfate hexahydrate [Fe(NH4)
(SO4)2.6H2O]. Dissolve it in sufficient
amount of water containing 2.5 mL of concentrated
sulphuric acid and complete to 100 mL by water
phenanthroline: dissolve 10 mg of the reagent -1,10 .2
in sufficient amount of water
containing drops of concentrated sulphuric acid and
.complete to 100 mL by water
Ascorbic acid: dissolve 1 g ascorbic acid in 100 mL .3
.distilled water
Sodium acetate 1M: dissolve a suitable amount in .4
.100 mL water
Acetic acid 1M: dilute about 6 mL of glacial acetic .5
.acid to 100 mL by water
Acetic acid-sodium acetate buffer (PH=4.5): mix .6
65 mL of 1M acetic acid and 35 mL of 1M sodium
acetate
:Preparation of standard solutions

Place one tablet (or 1 mL) of an iron-containing vitamin (note which .1
brand you use and the nominal mass of iron per tablet) in a 100-mL
beaker and boil gently on a hot plate (in a fume hood) with 25 mL of 6 M
.HCl for 15 min
Filter the solution directly into a 100-mL volumetric flask using .2
qualitative filter paper.Wash the beaker and filter paper several times
with small portions of water to complete aquantitative transfer. If some
insoluble bits make it through the filter paper, re-filter your solution
before proceeding
Allow the solution to cool, dilute to the mark, and mix well. (Note that .3
is important to let the solution cool before diluting, since volumetric flask
marks are accurate only at room temperature.)
Dilute 5.00 mL of this solution to 100.0 mL in a fresh volumetric flask. .4
If the label
indicates that the tablet contains <15 mg of Fe, use 10.00 mL instead of
5.00 mL. This“twice-diluted” solution is the unknown solution you will
use. While your original unknown solution will likely appear green or
yellow (depending on the dyes used in the tablet), your twice-diluted
solution will be clear
absorbance Wave
:Calculations length
0.073 360
=Weight of Fe(||) at complex ferrous ammonium sulfate hexahydrate •
0.09 380
wt = m.wt * m*v
0.206 400
gm 19.6 = 10-3*100* 0.5*392 =
0.33 420
preparation of sodium acetate •
0.416 440
W.t =M.W.T *M.V
0.486 460
M.W.T = =82gm/mol
0.554 480
Wt =82*100*10-³* 1 0.585 500
Wt=8.2gm 0.547 520
Preparation of ascorbic acid • 0.268 540
Weight of 1 gm in 100 ml D.W 0.086 560
Preparation of acetic acid 1M • 0.033 580
Dilute 6 ml of acetic acid to 100 ml by water 0.011 600
Preparation of acid sodium acetate buffer • 0.004 620
Mix 65 ml of 1M acetic acid and 35 ml of 1M sodium acetate
:Preparation of 1,10 phenanthroline •
Dissolve of 1 gm in 100 ml D.w
And withdraw 1 ml from it and dilute in 100 ml d.w
Lamda max = 500 nm
Absorbance Vs wave length

0.7
0.6
0.5
Abcorbance
0.4
0.3
0.2
0.1
0
300 350 400 450 500 550 600 650
wave length
Absorbance Vs Concetration
0.7
0.6
0.5
Absorbance
0.4
0.3
0.2
0.1
0
0 1 2 3 4 5 6
concentration
Absorbance concentration
0.159 0.5
0.122 0.75
0.211 1.25
0.363 2.5
0.461 3.75
0.615 5

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‫(‪)spectrophotometric‬‬

‫االسم ‪ :‬كيرلس عاطف جورجي‬

.measuring the intensity of light relative to wavelength

:Preparation of standard solutions

Lamda max = 500 nm

Absorbance Vs wave length

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