CHEM340

Tutorial: Quantitative methods of analysis.

Question 1
The following emission data was recorded for the ICP analysis for phosphorus standards.

[ P], mg L-1 Emission Intensity

2130 0.048
4260 0.110
6400 0.173
8530 0.230

When the emission data is plotted against the concentration of phosphorous, the intensity can be expressed
by the equation equation: Emission Intensity = 2.85x10-5[P] - 0.012.

a) Deduce the calibration sensitivity of the ICP method.

To determine the purity of a sample of Na2HPO4, a 2.469 g sample is dissolved and diluted to volume in a
100 mL volumetric flask. Analysis of the resulting solution for phosphorous gives an emission intensity of
0.135.
b) What is the percentage purity of the Na2HPO4?
c) Calculate the emission intensity of phosphorous for a sample that is 99% Na2HPO4 pure.

Question 2

The %(wt/wt) concentration of H2O in a methanol reagent was determined by GC, using a capillary column
coated with a non-polar stationary phase using an appropriate detector. A series of %( wt/wt) H2O-in-
methanol calibration standards gave the following results.

% (w/w) H2O (arb. units) Peak Height

0.00 1.15
0.014 2.74
0.047 6.33
0.095 11.58
0.176 20.43
0.290 32.97

Plotting the data gave the following equation: Peak height = 109.69 x %(wt/wt)H2O - 1.15

a) What is the %(wt/wt)H2O in a sample giving a peak height of 8.63?

The %(wt/wt) H2O in a freeze-dried antibiotic is determined in the following manner. A 0.175 g sample is
placed in a vial along with 4.500 g of methanol. Water in the sample extracts into the methanol. Analysis of
the extract gave a peak height of 13.66.
b) What is the %(wt/wt) H2O in the antibiotic? Show all your working.

Question 3
A fragrance mixture was prepared containing 10.0 ppm of an ester standard and 15.0 ppm of an appropriate
internal standard. A sample containing an unknown amount of the ester was treated in the same manner and
analyzed by gas chromatography. Measured data is summarized in the Table given below.

Peak area ( ester) Peak area (Internal standard)

10 ppm standard ester + 15 ppm internal 155 000 233 000
standard
Sample containing the ester fragrance 274 000 198 000
+ 15 ppm internal standard

a) Calculate the concentration (ppm) of the ester in the sample.

b) Why is the internal standard added to the ester standard as well as the sample?
c)
(i) Describe how the calibration could have been done using a multi-point-calibration approach.
(ii) Sketch the expected calibration graph and write down the general calibration equation derived
from it.
(iii) Show on the calibration graph how the concentration of the ester of an unknown sample can be
determined.

Question 4

A 0.100 g sample of groundnuts contaminated with aflatoxin B1 was extracted into an organic layer and the
extract transferred to a 250.0 mL volumetric flask and diluted to volume. Two 10.0 mL aliquots of the sample
were drawn out and further treated as follows:

Absorbance readings of sample and the sample spiked with an aflatoxin B1 standard.
Volume of sample (mL) Volume of a 1.00 ug L-1 Final volume (mL) Fluorescence
aflatoxin B1 standard (mL) intensity signal
10.00 0 25. 00 0.235
10.00 10 25.00 0.502

Calculate the concentration (ug kg-1) of aflatoxin B1 in the groundnuts sample. Show all your working.
Question 5

To determine the concentration of a certain antioxidant in a sample, a standard addition was performed. A
5.00 mL portion of the sample was analyzed and then successive 0.10 mL spikes of a 600.0 μg L-1 of an
antioxidant standard were added, analyzing the solution after each addition. The results are summarized in
the Table given below.

Absorbance readings of the solution after each addition of 0.10 mL spike of a 600.0 μg L-1 standard.
Volume (mL) of sample after at each addition of Volume of a 600 μg L-1 standard Absorbance
standard.
5.00 0.00 0.119
5.10 0.10 0.231
5.20 0.20 0.339
5.30 0.30 0.442

When the absorbance of the solution after each addition was plotted against the volumes of the added
standard, the following data were recorded. Analyze the data and use it to determine the concentration of the
antioxidant (μg L-1) in the sample.

0.45

0.40

0.35
Absorbance units

0.30

0.25

0.20

0.15

0.10

0.05

0.00
0.00 0.05 0.10 0.15 0.20 0.25 0.30

Volume of standard (mL)

Linear Regression for Data:

y=c+m*x
Parameter Value
--------------------------------
c 0.121
m 1.077
--------------------------------

Question 6

To analyze for vitamin A in a cereal, a 10.0 g sample of cereal is placed in a 250 mL Erlenmeyer flask along
with 1 g of sodium ascorbate, 40 mL of ethanol, and 10 mL of 50% (w/v) KOH. After refluxing for 30 min,
60 mL of ethanol is added, and the solution is cooled to room temperature. Vitamin A is extracted using three
100 mL portions of hexane. The combined portions of hexane are evaporated, and the residue containing
vitamin A is transferred to a 5 mL volumetric flask and diluted to volume with methanol. A standard addition
is prepared in a similar manner using a 10.0 g sample of the cereal and spiking it with 0.0200 mg of vitamin A.
Injecting the sample and standard addition into the HPLC gives peak areas of 6.77 x 103 and 1.32 x 104,
respectively.
Report the content of vitamin A in the sample in milligrams/100 g cereal.
Question 7

You are to calibrate a UV-Visible spectrophotometer for the analysis of trace amounts of nitrates (NO3-) in
50 mL water samples drawn at different sampling points from an inland lake. You gathered that the
concentration of nitrates in the lake samples ranges from 10 mg/L to about 40 mg/L.

You have a KNO3 primary standard for calibrating the instrument.

a) Calculate the mass of KNO3 (Mt = 101,11 g mol-1) that should be weighed and dissolved and made up to
1.0 L in order to prepare a 100.0 mg/L NO3- (Mt = 62,10 g mol-1) stock solution of nitrate.

In order to quantity the nitrates, a colour forming (derivatizing) reagent is added to the dissolved solution of
the primary standard before it is topped to 1L and subsequently diluted. The same addition is done to the
lake water samples.
During an instrument brief from the technician, you learnt that at least five working standards including a
reagent blank should be brought to the machine per group for the calibration of the machine. He advises you
that the linear range for nitrate calibration normally spans up to 50.00 mg/L. The wavelength of 540 nm is
going to be used for the analysis since it offers better sensitivity. He has already done a preliminary detection
limit experiment for you using a sample blank (a sample assumed to be free from a NO 3- ion which is being
analyzed) taken from a double distilled water. The detection limit had been estimated to be 0.08 mg/L.

b) Suggest a set of nitrate working standards which may be prepared for theproper calibration of the
machine.
Showing all sample calculations, detail how you would prepare the standards assuming that you have all
the conventional pipettes, the I.0 L and 6 x 100 mL volumetric flasks at your disposal. You may need to
draw up a summary table to show the amounts of stock solution pipette, the dilution factors involved
and the final concentrations of each of the prepared standard.

When you were almost done with the preparation of the working standards, a friend walk across to you and
thinks that instead of following the procedure as specified in the manual (preparing 100 mg/L of NO 3-) it is
more convenient to weigh the mass of KNO3 salt that will afford a 50 mg/L stock solution in NO 3- ions and it
would save time and reduces errors associated with large dilution factors in the subsequent preparations of
the working standards.
c)
(i) Showing all working, criticize your friend’s suggestion to prepare the 50 mg/L stock solution in NO3-
ions directly from the nitrate primary salt standard.

(ii) Reflecting on the briefing from the technician, what is meant by sensitivity and linear dynamic range? In
what ways are they important factors in quantification by instrumental analysis?

(iii) Briefly describe how the technician estimated the detection limit of the instrument for the nitrate
determination.
You are further told that linear regression of the calibration equation is done promptly by an online program
on the machine.
d)
(i) Explain the principles of linear regression through the least-square method.

Your regression results read:

Linear regression: Abs unit = B*{[NO3-]ppm} + C
Output Standard error

B 0.018 ppm-1 ±0.004

C 0.0044 abs units ±0.003
Spillway sample (diluted three 0.677 abs units ±0.001
times)
R2 = 99.998%; n = 5

(ii) Calculate the concentration of the spillway sample and its uncertainty.

(iii) Calculate the 99% confidence interval of the slope of the calibration curve.

(iv) Deduce the reading (abs. units) of your top working standard (from b)) from the calibration equation.

(v) What is the significance of the value of R2 given as a footnote in the regression data?