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Real concentration:
C1*V1=C2*V2
C1-concentration before diluting
V1-25 ml pipette
C2-concentration after diluting
V2-50 ml flask
( x Xi )2
n1
chemical
quinineHCl2H2O
acid micture
(HCl+H3PO4)
sample tonic water
brand name
Acros Organics
N.A.
lot number
163720250
N.A.
concentration
99%
1 M (1:1)
date
N.A.
N.A.
Royal Club
0042L26805
N.A.
04.2014.
Equipment(table 2)
equipment
spectrophotometer
scale
brand name
Shimadzu
Sartorius
type
UV-1800
M-Power
registration number
Han-05125548
Han-05125542/00391
4.2. Procedure:
1. Preparing the quinine stock solution:
1.1 Weigh carefully about 155 mg of quinineHCl2H2O and dissolve in 50 ml acid mixture.
1.2 Transfer the solution to a 250 ml volumetric flask, top up with demineralised water to the line
and homogenize.
2. Calibration series:
3.1.Using the stock solution quinineHCl2H2O make the quinine standard solution with a final
concentration of 10-20-30-40-50 mg quinine/l in volumetric flasks of 50 ml.
3.2.To each volumetric flask add 12.5 ml acid mixture.
3.3.Top up the volumetric flasks to the mark and homogenize.
4. Preparing the sample solution:
4.1.Pipette (in triplo) 25 ml of analysis material in a 50 ml volumetric flask. Add 12.5 ml of the acid
mixture and top up to the line and homogenize.
5. Measurements:
5.1.Using a solution with the concentration of 40 mg quinine/l take a spectrum of 300 up to 400 nm
to determine the maximum wavelength.
5.2.Adjust the spectrophotometer to this fixed wavelength.
5.3.Measure the absorbance of all standards and samples.
5.4. Using the standard solutions, calculate a calibration line and determine the concentration of
quinine in tonic with the use of this line. Do not forget the dilution of the sample. Calculate
using the actual concentration of the quinine standard solution.
5. Results
The quinine calibration standards data are presented in table 3 and 4. A plot of Concentration against Absorbance of
tonic water samples is presented in Graph 1. This plot has a regression coefficient (R2) of 0.999.
Measurement Data-table 3
VquinineHCl2H2O pipetted
(l)
10.1342
20.2684
30.4026
40.5368
50.671
2
Absorbance
0.157
0.319
0.481
0.650
0.820
max = 345.25 nm
Absorbance(max)=0.7
52
Measurement Data-table 4
1
28.21
0.442
56.63
Absorbance
Real concentration (mg/ml)
2
28.06
0.438
56.125
3
28.44
0.444
56.875
0.5
Linear (Absorbance)
Absorbance 0.4
Linear (Absorbance)
0.3
0.2
0.1
0
0 10 20 30 40 50 60
Concentration (mg/l)
Calculation examples:
example 1
i/MquinineHCl2H O=
2
620 mg/l
396.91 g/mol
= 1.562 mol/l
example 2
Volume of stock solution to be pipetted:
C1*V1=C2*V2
500* V1=10*50
V1= 1 ml
example 3
Concentration of calibration series(standard solutions):
C1*V1=C2*V2
506.71*1= C2*50
x=
s=
s=
s=
s=
s=
( x Xi )2
0.00701+0.1737+ 0.1109
2
n1
2
( x Xi ) + ( x Xi ) + ( x Xi )
31
( 56.5418656.6256
) + ( 56.5418656.125 )2 + ( 56.5418656.875
)
0.29161
2
s=0.3818
6.
Discussion/Conclusion
From the completion of 5 standard solutions of quinine hydrochlorid dihydrate and 3 sample solutions of tonic water, the
amount of quinine was determined to be about 56 ppm. This value was calculated via Concentration against Abrosbance
graph which was plotted in Excel. It seems reasonable because it falls under the range given by US Food and Drug
Administration, which limits it to 83 ppm. As seen in the spectrophotometry results, the maximum absorbance
wavelength was 345.25 nm. This matches hypotheses, made prior to the experiment, that quinine absorbance peaks
around 350 nm. Overall, this data met the objectives for this experiment and it is of good quality. The regression
coefficient of 0.999 is very high and shows that the best-fit line is of good quality and represents the data well. This is
important because we used that slope in later analysis, so this proves the quality of the calculations as a whole. There
were some possible sources of error in this experiment due to impurities of glassware and cuvettes, random errors or
imprecise measurements which may have altered the results. The calculated standard deviation confirms that the random
error was very small.
i =
mquinineHCl2 H 2 O
154.9 mg
=
= 619.6 mg/l
V flask
0.25 l
i/MquinineHCl2H O=
2
619.6 mg/l
396.91 g/mol